Get 20M+ Full-Text Papers For Less Than $1.50/day. Start a 14-Day Trial for You or Your Team.

Learn More →

Long Non-Coding RNA (LncRNA) CASC15 Is Upregulated in Diabetes-Induced Chronic Renal Failure and Regulates Podocyte Apoptosis

Long Non-Coding RNA (LncRNA) CASC15 Is Upregulated in Diabetes-Induced Chronic Renal Failure and... CLINICAL RESEARCH e-ISSN 1643-3750 © Med Sci Monit, 2020; 26: e919415 DOI: 10.12659/MSM.919415 Received: 2019.08.14 Long Non-Coding RNA (LncRNA) CASC15 Is Accepted: 2019.11.05 Available online: 2020.01.21 Upregulated in Diabetes-Induced Chronic Renal Published: 2020.02.13 Failure and Regulates Podocyte Apoptosis Authors’ Contrib ution: BCDEF Xiaohua Qin Department of Nephrology, Second Affiliated Hospital of Nanchang University, Study Design A Nanchang, Jiangxi, P.R. China BCD Shuying Zhu Data Collection B BC Yanxia Chen Statistical Analysis C Data Interpr etation D BC Dan Chen Manuscript Preparation E B Weiping Tu Literature Search F ABCDEF Hongchang Zou Funds Collection G Corresponding Author: Hongchang Zou, e-mail: vcarnmzqgmi56@163.com Source of support: Departmental sources Background: CASC15 has been recently characterized as an oncogenic lncRNA. This study aimed to investigate the role of CASC15 in diabetic patients complicated with chronic renal failure (DCRF). Material/Methods: Levels of CASC15 in plasma derived from 3 groups of participants were measured by qPCR and compared by ANOVA and Tukey test. The interaction between CASC15 and miR-34c was analyzed by performing cell trans- fections. Cell apoptosis assay was performed to analyze the effects of transfections on the apoptosis of CIHP-1 cells (podocytes). Results: We found that CASC15 in plasma was upregulated in DCRF compared with diabetic patients (no obvious com- plications) and healthy controls. Upregulation of CASC15 distinguished DCRF patients from healthy controls and diabetic patients. High D-glucose environment induced the upregulation of CASC15 in cells of the human podocyte cell line CIHP-1. Overexpression of CASC15 did not affect miR-34c in CIHP-1 cells, but bioinformatics analysis showed that CASC15 can sponge miR-34c. Overexpression of CASC15 led to an increased apoptotic rate of CIHP-1 cells, and miR-34c overexpression led to a decreased apoptotic rate of CIHP-1 cells. In addition, CASC15 overexpression attenuated the effects of miR-34c overexpression on cell apoptosis. Conclusions: Therefore, CASC15 is upregulated in DCRF patients and promotes the apoptosis of podocytes by sponging miR-34c. Our study adds to our understanding of the pathogenesis of DCRF and suggests that CASC15 could serve as a potential therapeutic target of DCRF. MeSH Keywords: Apoptosis • Kidney Failure, Chronic • MicroRNAs Full-text PDF: https://www.medscimonit.com/abstract/index/idArt/919415 1879 — 4 20 Indexed in: [Current Contents/Clinical Medicine] [SCI Expanded] [ISI Alerting System] This work is licensed under Creative Common Attribution- [ISI Journals Master List] [Index Medicus/MEDLINE] [EMBASE/Excerpta Medica] e919415-1 NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0) [Chemical Abstracts/CAS] Qin X. et al.: LncRNA CASC15 regulates renal cell apoptosis CLINICAL RESEARCH © Med Sci Monit, 2020; 26: e919415 Background complications. Exclusion criteria were: 1) complications were observed, and 2) other severe diseases were observed. The par- Diabetes is one of the most prevalent diseases in both develop- ticipants were informed about the details of the study and all ing and developed countries [1]. With changes in modern soci- of them signed written informed consent. ety, the number of people living with this disease has doubled during the past 20 years [1]. It is predicted that this number Plasma specimens will continuously increase at least until 2030 [2]. Currently, dia- betes is a major public health burden [3]. Compared to healthy Before the initiation of therapies, 5 ml of blood was extracted adults without diabetes, a 50% higher risk of death was ob- under fasting conditions. To prepare plasma samples, blood served in adults with diabetes, largely due to the fact that al- was placed into EDTA tubes, which were centrifuged for 20 min most all diabetic patients will develop complications during the at 1200 g to collect the supernatant. progression of disease [4]. At present, prevention of diabetic complications is still critical for long-time survival of patients [5]. Podocytes The long-term high-glucose environment in diabetic patients The human podocyte cell line CIHP-1 (Ximbio) was used in this can seriously affect functions of the most important organs study. The reason we used podocytes is that glomerular func- and cause development of multiple diabetic complications, tion requires the involvement of podocytes, and chronic renal such as diabetic chronic renal failure (DCRF) [6]. Under high- failure results in dysfunctional podocytes. CIHP-1 cells were glucose conditions, wound-healing of kidney tissue is usually cultivated in a mixture of 10% FBS and 90% RPMI-1640 under unsuccessful and chronic renal failure is common [6]. DCRF the conditions of 37°C, 95% humidity, and 5% CO . For high- patients usually suffer from multiple functional disorders of glucose treatment, we added 5, 10, 20, and 30 mM D-glucose the kidney, such as the apoptosis of renal cells [7]. Therefore, (Sigma-Aldrich) to the medium, and cells were harvested at inhibition of renal apoptosis is a potential therapeutic ap- 12, 24, and 48 h after the initiation of treatment. proach for clinical treatment. It has been reported that DCRF requires the involvement of multiple genetic factors [8,9]. In a Transient cell transfections recent study, Liu et al. showed that miR-34c can target Notch signaling pathways to inhibit the apoptosis of podocytes in- The CASC15 expression vector was constructed using pcDNA3.1 duced by high glucose [10]. CASC15 is a well-studied onco- vector (Sangon, Shanghai, China). Negative control (NC) miRNA genic lncRNA; however, the role of CASC15 in renal injuries is and miR-34c mimic were from Sangon. CIHP-1 cells were unknown [11]. In our preliminary experiments, bioinformatics counted. The transient transfections were performed using lipo- analysis revealed the potential targeting effect between CASC15 fectamine 2000 reagent (Invitrogen, USA) to transfect 15 nM and miR-34c. This study aimed to investigate the exact inter- vectors (empty vector as NC group) or 35 nM miRNAs (NC miRNA actions between CASC15 and miR-34c in DCRF. as NC group) into 2×10 cells. The following experiments were performed using cells harvested at 24 h after transfection. For all transfections, control (C) cells were untransfected cells. Material and Methods RNA extractions and qPCR Research subjects Total RNAs in 0.3 ml plasma and 2×10 cells were extracted Research subjects of the present study included 50 DCRF pa- using RNAzol (Sigma-Aldrich). To harvest miRNAs, 85% etha- tients (DCRF group, 26 males and 24 females; age 36–67 nol was used to precipitate all RNA samples. All RNA samples years, mean age 51.9±6.0 years), 50 diabetic patients with- were digested with DNase I for 80 min at 37°C to completely out obvious complications (Diabetes group, 26 males and 24 remove genomic DNA. The digested DNA samples were re- females; age 35–68 years, mean age 52.3±6.5 years), and 50 verse-transcribed into cDNA using Tetro Reverse Transcriptase healthy volunteers (Control group, 26 males and 24 females; (Bioline), and cDNA was used as a template to prepare qPCR age 36–68 years, mean age 52.0±6.3 years). All participants mixtures using Brilliant III Ultra-Fast SYBR Green QPCR Master were selected at the Second Affiliated Hospital of Nanchang Mix (Agilent Technologies). With 18S rRNA as an endoge- University between April 2016 and April 2018. All patients were nous control, the expression levels of CASC15 were detected. diagnosed according to the criteria proposed by the Chinese To measure the expression levels of miR-34c, all steps were Medical Association. This study was approved by the hospi- completed using the All-in-OneTM miRNA qRT-PCR Detection tal Ethics Committee. Patient inclusion criteria were: 1) newly Kit (GeneCopoeia). U6 was used as the endogenous control. –DDCT diagnosed cases, 2) no therapies were performed within 100 The 2 method was used to process all data. All PCR reac- days before admission, and 3) CRF caused by diabetes without tions were performed in triplicate. Indexed in: [Current Contents/Clinical Medicine] [SCI Expanded] [ISI Alerting System] This work is licensed under Creative Common Attribution- [ISI Journals Master List] [Index Medicus/MEDLINE] [EMBASE/Excerpta Medica] e919415-2 NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0) [Chemical Abstracts/CAS] Qin X. et al.: LncRNA CASC15 regulates renal cell apoptosis CLINICAL RESEARCH © Med Sci Monit, 2020; 26: e919415 Control (n=50) Control (n=50) Control (n=50) B C 100 100 80 80 60 60 40 40 Sensitivity% Sensitivity% 20 20 Identity% Identity% 0 0 02040 60 80 100 02040 60 80 100 100%-Speci city% 100%-Speci city% Figure 1. Plasma CASC15 was upregulated in DCRF and showed diagnostic values. Levels of CASC15 in plasma derived from 3 groups of participants (DCRF, Diabetes, and Control) were measured by qPCR and compared by ANOVA (one-way) and Tukey test (A). In ROC curve, true-positive cases were DCRF patients and true-negative cases were either diabetic patients (B) or healthy controls (C). Experiments were performed in 3 replicates and mean values are presented, * p<0.05. Cell apoptosis analysis different patient and cell groups by performing ANOVA (one- way) and Tukey test. The diagnostic analysis was performed CIHP-1 cells were harvested at 24 h after transfection and using ROC curves. In ROC curves, true-positive cases were DCRF counted. To prepare a single-cell suspension, 4×10 cells were patients and true-negative cases were either diabetic patients mixed with 1 ml of the cell culture medium. Cells were trans- or healthy controls. p<0.05 was statistically significant. ferred to a 6-well plate with 2 ml cell suspension per well, fol- lowed by the addition of 30 mM D-glucose. Cells were cultivat- ed for 48 h under the aforementioned conditions. After that, Results cells were washed with PBS, and 0.25% trypsin digestion was performed. Finally, cells were stained with Annexin V-FITC and Plasma CASC15 was upregulated in DCRF and showed propidium iodide (PI) for 20 min in the dark, and flow cytome - diagnostic values try was then performed to separate apoptotic cells. Levels of CASC15 in plasma derived from 3 groups of partici- Statistical analysis pants (DCRF, Diabetes, and Control) were measured by qPCR and compared by one-way ANOVA and Tukey test. Comparing The mean values of 3 biological replicates were calculated and to the Control group, significantly higher plasma levels of were used in all data analyses. Differences were explored among CASC15 were observed in the DCRF group but not in the Indexed in: [Current Contents/Clinical Medicine] [SCI Expanded] [ISI Alerting System] This work is licensed under Creative Common Attribution- [ISI Journals Master List] [Index Medicus/MEDLINE] [EMBASE/Excerpta Medica] e919415-3 NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0) [Chemical Abstracts/CAS] 100% Sensitivity Relative CASC15 level 100% Sensitivity Qin X. et al.: LncRNA CASC15 regulates renal cell apoptosis CLINICAL RESEARCH © Med Sci Monit, 2020; 26: e919415 Figure 2. Upregulation of CASC15 was induced by high glucose in CIHP-1 cells. CIHP-1 cells were treated with 5, 10, 20, 5 5 mM 10 mM and 30 mM D-glucose for 12, 24, and 48 h, followed by 20 mM measuring the expression levels of CASC15 by qPCR. 30 mM Data were compared by ANOVA (one-way) and Tukey test. Experiments were performed in 3 replicates and mean values are presented, * p<0.05. 0 12 24 48 * * 5 * 8 * C NC CASC15 C NC CASC15 1.5 1.5 1.0 1.0 0.5 0.5 0.0 0.0 C NC CASC15 C NC CASC15 2 2 0 miR-34c 5’ - AG GG UUGC-3’ GCA UGUAGUUA CUGA CASC15 UGU ACAUCAGU GACU 3’-AAU . . . AAUUG AG G CCCAU . . . AGG-5’ 510 491 Figure 3. CASC15 can sponge miR-34c. CIHP-1 cells were transfected with the CASC15 expression vector and miR-34c mimic. Overexpression of CASC15 and miR-34c was confirmed at 24 h after transfection by qPCR ( A). The interaction between CASC15 and miR-34c was analyzed by qPCR (B). IntaRNA was used to predict the interaction between miR-34c and CASC15 (C). Experiments were performed in 3 replicates and mean values are presented, * p<0.05. Indexed in: [Current Contents/Clinical Medicine] [SCI Expanded] [ISI Alerting System] This work is licensed under Creative Common Attribution- [ISI Journals Master List] [Index Medicus/MEDLINE] [EMBASE/Excerpta Medica] e919415-4 NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0) [Chemical Abstracts/CAS] Relative expression level of CASC15 Relative miR-34c level Relative CASC15 level Relative CASC15 level Relative miR-34c level Qin X. et al.: LncRNA CASC15 regulates renal cell apoptosis CLINICAL RESEARCH © Med Sci Monit, 2020; 26: e919415 C NC miRNA pcDNA3.1 4 4 4 10 10 10 3 3 3 10 10 10 2 2 2 PI 10 PI 10 PI 10 1 1 1 10 10 10 0 0 0 10 10 10 0 1 2 3 4 0 1 2 3 4 0 1 2 3 4 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 miR-34c CASC15 CASC15+miR-34c 4 4 4 10 10 10 3 3 3 10 10 10 2 2 2 PI 10 PI 10 PI 10 1 1 1 10 10 10 0 0 0 10 10 10 0 1 2 3 4 0 1 2 3 4 0 1 2 3 4 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 C NC CASC15 CASC15 CASC15 CASC15 Figure 4. CASC15 attenuated the effects of miR-34c on cell apoptosis. Cell apoptosis assay was performed to analyze the effects of transfections on apoptosis of CIHP-1 cells. Experiments were performed in 3 replicates and mean values are presented, * p<0.05. Diabetes group (Figure 1, p<0.05). The diagnostic value of plas- Upregulation of CASC15 was induced by high glucose in ma CASC15 for DCRF was analyzed by ROC curve. AUC>0.65 CIHP-1 cells indicates potential diagnostic value. With diabetic patients as true-negative cases, the AUC was 0.97 (95% confidence inter - CIHP-1 cells were treated with 5, 10, 20, and 30 mM D-glucose val: 0.95–0.99; standard error: 0.013). With healthy controls as for 12, 24, and 48 h, followed by measuring the expression lev- true-negative cases, the AUC was 0.97 (95% confidence inter - els of CASC15 by qPCR. As shown in Figure 2, expression levels val: 0.95–1.00; standard error: 0.013). AUC>0.65 indicated di- of CASC15 in CIHP-1 cells were upregulated by D-glucose in a agnostic values. Therefore, plasma CASC15 can serve as a di- dose- and time-dependent manner (Figure 2, p<0.05). However, agnostic marker for DCRF. 5 mM D-glucose failed to affect CASC15 expression, possibly because 5 mM is within the normal range. Indexed in: [Current Contents/Clinical Medicine] [SCI Expanded] [ISI Alerting System] This work is licensed under Creative Common Attribution- [ISI Journals Master List] [Index Medicus/MEDLINE] [EMBASE/Excerpta Medica] e919415-5 NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0) [Chemical Abstracts/CAS] Apoptotic cell percentage Qin X. et al.: LncRNA CASC15 regulates renal cell apoptosis CLINICAL RESEARCH © Med Sci Monit, 2020; 26: e919415 CASC15 can sponge miR-34c environment led to significantly upregulated expression of CASC15 in podocytes. The possible reason is that the upregu- CIHP-1 cells were transfected with the CASC15 expression vector lated CASC15 in podocytes fails to affect the plasma levels of and miR-34c mimic. Expression levels of CASC15 and miR-34c CASC15 in diabetic patients. With the development of DCRF, were measured at 24 h after transfection. Compared to the C CASC15 is further upregulated and more CASC15 is released and NC groups, expression levels of CASC15 and miR-34c were into blood, resulting in increased plasma levels of CASC15. significantly increased after transfections (Figure 3A, p<0.05). However, overexpression of CASC15 and miR-34c failed to af- This study showed that the upregulation of plasma CASC15 fect the expression of each other (Figure 3B). Bioinformatics could be used to assist in diagnosis of DCRF, but the accura- analysis using IntaRNA showed that miR-34c can bind CASC15 cy remains to be further analyzed. We showed that CASC15 from position 492 to 509 (Figure 3C). can interact with miR-34c, while overexpression experiments showed that CASC15 and miR-34c have no effects on the ex - CASC15 attenuated the effects of miR-34c on cell pression of each other. Therefore, it appears that miR-34c apoptosis does not target CASC15. It is known that lncRNAs can sponge miRNAs to attenuate their functions [16,17]. The main role of Cell apoptosis assay was performed to analyze the effects of a miRNA sponge is to inhibit the roles of miRNA. This study transfections on apoptosis of CIHP-1 cells. Compared to the revealed that CASC15 attenuated the effects of miR-34c on C and NC groups, overexpression of CASC15 led to increased cell apoptosis. Therefore, CASC15 can sponge miR-34c to pro- apoptotic rate of CIHP-1 cells and miR-34c overexpression led mote the apoptosis of podocytes. to a decreased rate. In addition, CASC15 overexpression at- tenuated the effects of miR-34c overexpression on cell apop - CASC15 is dysregulated in multiple clinical disorders, such as dif- tosis (Figure 4, p<0.05). ferent types of cancers and cardiac hypertrophy [11,15,18–20]. Therefore, the use of CASC15 as a diagnostic marker for DCRF should exclude the possibility of these clinical disorders. In ad- Discussion dition, this study was limited by its small sample size. Studies with larger sample sizes are needed to confirm our conclusions. In this study, we investigated the functions of CASC15 in DCRF and also analyzed the clinical value. We found that CASC15 was upregulated in DCRF and can sponge miR-34c to promote Conclusions the apoptosis of podocytes. In conclusion, CASC15 was upregulated in DCRF and can sponge Previous studies have revealed many lncRNAs involved in the miR-34c to promote the apoptosis of podocytes. Our study pro- development of renal disorders. For instance, lncRNA Erbb4-IR vides novel insights into the molecular mechanism of DCRF can target miR-29b to promote renal injury under a high-glu- and suggests potential therapeutic targets. cose environment [12]. lncRNA MALAT1 was reported to be up- regulated in diabetic patients, and downregulation of MALAT1 Availability of data and materials can promote recovery of renal functions in a rat model [13]. In another study, Wang et al. showed that p53 can downregu- The analyzed datasets generated during the study are avail- late lncRNA ZEB1-AS1 to participate in renal fibrosis in patients able from the corresponding author on reasonable request. with diabetic nephropathy [14]. CASC15 is a well-characterized oncogenic lncRNA [11,15]. CASC15 is upregulated in cancers, Ethics approval and consent to participate and the overexpression of CASC15 promotes cancer progres- sion by regulating cancer cell behaviors, such as inhibition of The present study was approved by the Ethics Committee of cell apoptosis [15]. The present study is the first to report the the Second Alffi iated Hospital of Nanchang University. The re - upregulation of CASC15 and the increased cell apoptosis rate search was carried out in accordance with the World Medical after CASC15 overexpression. Therefore, CASC15 can contrib- Association Declaration of Helsinki. All patients and healthy ute to the development of DCRF. Our study also showed that volunteers provided written informed consent prior to their CASC15 can have opposite roles in regulating cell apoptosis inclusion in the study. in different diseases. Conflict of interests Our study did not observe the upregulation of CASC15 in plas- ma derived from diabetic patients. However, a high-glucose None. Indexed in: [Current Contents/Clinical Medicine] [SCI Expanded] [ISI Alerting System] This work is licensed under Creative Common Attribution- [ISI Journals Master List] [Index Medicus/MEDLINE] [EMBASE/Excerpta Medica] e919415-6 NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0) [Chemical Abstracts/CAS] Qin X. et al.: LncRNA CASC15 regulates renal cell apoptosis CLINICAL RESEARCH © Med Sci Monit, 2020; 26: e919415 References: 1. Zimmet P, Alberti KG, Magliano DJ et al: Diabetes mellitus statistics on prev- 12. Sun SF, Tang PMK, Feng M et al: Novel lncRNA Erbb4-IR promotes diabetic alence and mortality: facts and fallacies. Nat Rev Endocrinol, 2016; 12(10): kidney injury in db/db mice by targeting miR-29b. Diabetes, 2018; 67(4): 616–22 731–44 2. Rowley WR, Bezold C, Arikan Y et al: Diabetes 2030: Insights from yester- 13. Wu D, Cheng Y, Huang X et al: Down-regulation of lncRNA MALAT1 con- day, today, and future trends. Popul Health Manag, 2017; 20(1): 6–12 tributes to renal functional improvement after duodenal-jejunal bypass in a diabetic rat model. J Physiol Biochem, 2018; 74(3): 431–39 3. American Diabetes Association: Economic costs of diabetes in the US in 2017. Diabetes Care, 2018; 41(5): 917–28 14. Wang J, Pang J, Li H et al: lncRNA ZEB1-AS1 was suppressed by p53 for renal fibrosis in diabetic nephropathy. Mol Ther Nucleic Acids, 2018; 12: 741–50 4. Brownlee M: The pathobiology of diabetic complications: A unifying mech- anism. Diabetes, 2005; 54(6): 1615–25 15. Fernando TR, Contreras JR, Zampini M et al: The lncRNA CASC15 regulates SOX4 expression in RUNX1-rearranged acute leukemia. Mol Cancer, 2017; 5. Testa R, Bonfigli A, Genovese S et al: The possible role of flavonoids in the 16(1): 126 prevention of diabetic complications. Nutrients, 2016; 8(5): pii: E310 16. Liang WC, Fu WM, Wong CW et al: The lncRNA H19 promotes epithelial 6. Bos-Touwen I, Schuurmans M, Monninkhof EM et al: Patient and disease to mesenchymal transition by functioning as miRNA sponges in colorectal characteristics associated with activation for self-management in patients cancer. Oncotarget, 2015; 6(26): 22513–25 with diabetes, chronic obstructive pulmonary disease, chronic heart failure and chronic renal disease: A cross-sectional survey study. PLoS One, 2015; 17. Liu D, Li Y, Luo G et al: LncRNA SPRY4-IT1 sponges miR-101-3p to promote 10(5): e0126400 proliferation and metastasis of bladder cancer cells through up-regulating EZH2. Cancer Lett, 2017; 388: 281–91 7. Miranda-Díaz AG, Pazarín-Villaseñor L, Yanowsky-Escatell FG et al: Oxidative stress in diabetic nephropathy with early chronic kidney disease. J Diabetes 18. Yao XM, Tang JH, Zhu H et al: High expression of LncRNA CASC15 is a risk Res, 2016; 2016: 7047238 factor for gastric cancer prognosis and promote the proliferation of gas- tric cancer. Eur Rev Med Pharmacol Sci, 2017; 21(24): 5661–67 8. Ahlqvist E, Van Zuydam NR, Groop LC et al: The genetics of diabetic com- plications. Nat Rev Nephrol, 2015; 11(5): 277–87 19. Li C, Zhou G, Feng J et al: Up-regulation of lncRNA VDR/CASC15 induced by facilitates cardiac hypertrophy through modulating miR-432-5p/TLR4 axis. 9. Ma RCW: Genetics of cardiovascular and renal complications in diabetes. J Biochem Biophys Res Commun, 2018; 503(4): 2407–14 Diabetes Investig, 2016; 7(2): 139–54 20. Jing N, Huang T, Guo H et al: LncRNA CASC15 promotes colon cancer cell pro- 10. Liu XD, Zhang LY, Zhu TC et al: Overexpression of miR-34c inhibits high glu- liferation and metastasis by regulating the miR-4310/LGR5/Wnt/b-catenin cose-induced apoptosis in podocytes by targeting Notch signaling path- signaling pathway. Mol Med Rep, 2018; 18(2): 2269–76 ways. Int J Clin Exp Pathol, 2015; 8(5): 4525–34 11. He T, Zhang L, Kong Y et al: Long non-coding RNA CASC15 is up-regulat- ed in hepatocellular carcinoma and facilitates hepatocarcinogenesis. Int J Oncol, 2017; 51(6): 1722–30 Indexed in: [Current Contents/Clinical Medicine] [SCI Expanded] [ISI Alerting System] This work is licensed under Creative Common Attribution- [ISI Journals Master List] [Index Medicus/MEDLINE] [EMBASE/Excerpta Medica] e919415-7 NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0) [Chemical Abstracts/CAS] http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Medical Science Monitor : International Medical Journal of Experimental and Clinical Research Pubmed Central

Long Non-Coding RNA (LncRNA) CASC15 Is Upregulated in Diabetes-Induced Chronic Renal Failure and Regulates Podocyte Apoptosis

Qin, Xiaohua; Zhu, Shuying; Chen, Yanxia; Chen, Dan; Tu, Weiping; Zou, Hongchang
Medical Science Monitor : International Medical Journal of Experimental and Clinical Research , Volume 26 – Feb 13, 2020
Download PDF
7 pages

Loading next page...
 
/lp/pubmed-central/long-non-coding-rna-lncrna-casc15-is-upregulated-in-diabetes-induced-OK9uheCUbd

References (21)

Publisher
Pubmed Central
Copyright
© Med Sci Monit, 2020
ISSN
1234-1010
eISSN
1643-3750
DOI
10.12659/MSM.919415
Publisher site
See Article on Publisher Site

Abstract

CLINICAL RESEARCH e-ISSN 1643-3750 © Med Sci Monit, 2020; 26: e919415 DOI: 10.12659/MSM.919415 Received: 2019.08.14 Long Non-Coding RNA (LncRNA) CASC15 Is Accepted: 2019.11.05 Available online: 2020.01.21 Upregulated in Diabetes-Induced Chronic Renal Published: 2020.02.13 Failure and Regulates Podocyte Apoptosis Authors’ Contrib ution: BCDEF Xiaohua Qin Department of Nephrology, Second Affiliated Hospital of Nanchang University, Study Design A Nanchang, Jiangxi, P.R. China BCD Shuying Zhu Data Collection B BC Yanxia Chen Statistical Analysis C Data Interpr etation D BC Dan Chen Manuscript Preparation E B Weiping Tu Literature Search F ABCDEF Hongchang Zou Funds Collection G Corresponding Author: Hongchang Zou, e-mail: vcarnmzqgmi56@163.com Source of support: Departmental sources Background: CASC15 has been recently characterized as an oncogenic lncRNA. This study aimed to investigate the role of CASC15 in diabetic patients complicated with chronic renal failure (DCRF). Material/Methods: Levels of CASC15 in plasma derived from 3 groups of participants were measured by qPCR and compared by ANOVA and Tukey test. The interaction between CASC15 and miR-34c was analyzed by performing cell trans- fections. Cell apoptosis assay was performed to analyze the effects of transfections on the apoptosis of CIHP-1 cells (podocytes). Results: We found that CASC15 in plasma was upregulated in DCRF compared with diabetic patients (no obvious com- plications) and healthy controls. Upregulation of CASC15 distinguished DCRF patients from healthy controls and diabetic patients. High D-glucose environment induced the upregulation of CASC15 in cells of the human podocyte cell line CIHP-1. Overexpression of CASC15 did not affect miR-34c in CIHP-1 cells, but bioinformatics analysis showed that CASC15 can sponge miR-34c. Overexpression of CASC15 led to an increased apoptotic rate of CIHP-1 cells, and miR-34c overexpression led to a decreased apoptotic rate of CIHP-1 cells. In addition, CASC15 overexpression attenuated the effects of miR-34c overexpression on cell apoptosis. Conclusions: Therefore, CASC15 is upregulated in DCRF patients and promotes the apoptosis of podocytes by sponging miR-34c. Our study adds to our understanding of the pathogenesis of DCRF and suggests that CASC15 could serve as a potential therapeutic target of DCRF. MeSH Keywords: Apoptosis • Kidney Failure, Chronic • MicroRNAs Full-text PDF: https://www.medscimonit.com/abstract/index/idArt/919415 1879 — 4 20 Indexed in: [Current Contents/Clinical Medicine] [SCI Expanded] [ISI Alerting System] This work is licensed under Creative Common Attribution- [ISI Journals Master List] [Index Medicus/MEDLINE] [EMBASE/Excerpta Medica] e919415-1 NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0) [Chemical Abstracts/CAS] Qin X. et al.: LncRNA CASC15 regulates renal cell apoptosis CLINICAL RESEARCH © Med Sci Monit, 2020; 26: e919415 Background complications. Exclusion criteria were: 1) complications were observed, and 2) other severe diseases were observed. The par- Diabetes is one of the most prevalent diseases in both develop- ticipants were informed about the details of the study and all ing and developed countries [1]. With changes in modern soci- of them signed written informed consent. ety, the number of people living with this disease has doubled during the past 20 years [1]. It is predicted that this number Plasma specimens will continuously increase at least until 2030 [2]. Currently, dia- betes is a major public health burden [3]. Compared to healthy Before the initiation of therapies, 5 ml of blood was extracted adults without diabetes, a 50% higher risk of death was ob- under fasting conditions. To prepare plasma samples, blood served in adults with diabetes, largely due to the fact that al- was placed into EDTA tubes, which were centrifuged for 20 min most all diabetic patients will develop complications during the at 1200 g to collect the supernatant. progression of disease [4]. At present, prevention of diabetic complications is still critical for long-time survival of patients [5]. Podocytes The long-term high-glucose environment in diabetic patients The human podocyte cell line CIHP-1 (Ximbio) was used in this can seriously affect functions of the most important organs study. The reason we used podocytes is that glomerular func- and cause development of multiple diabetic complications, tion requires the involvement of podocytes, and chronic renal such as diabetic chronic renal failure (DCRF) [6]. Under high- failure results in dysfunctional podocytes. CIHP-1 cells were glucose conditions, wound-healing of kidney tissue is usually cultivated in a mixture of 10% FBS and 90% RPMI-1640 under unsuccessful and chronic renal failure is common [6]. DCRF the conditions of 37°C, 95% humidity, and 5% CO . For high- patients usually suffer from multiple functional disorders of glucose treatment, we added 5, 10, 20, and 30 mM D-glucose the kidney, such as the apoptosis of renal cells [7]. Therefore, (Sigma-Aldrich) to the medium, and cells were harvested at inhibition of renal apoptosis is a potential therapeutic ap- 12, 24, and 48 h after the initiation of treatment. proach for clinical treatment. It has been reported that DCRF requires the involvement of multiple genetic factors [8,9]. In a Transient cell transfections recent study, Liu et al. showed that miR-34c can target Notch signaling pathways to inhibit the apoptosis of podocytes in- The CASC15 expression vector was constructed using pcDNA3.1 duced by high glucose [10]. CASC15 is a well-studied onco- vector (Sangon, Shanghai, China). Negative control (NC) miRNA genic lncRNA; however, the role of CASC15 in renal injuries is and miR-34c mimic were from Sangon. CIHP-1 cells were unknown [11]. In our preliminary experiments, bioinformatics counted. The transient transfections were performed using lipo- analysis revealed the potential targeting effect between CASC15 fectamine 2000 reagent (Invitrogen, USA) to transfect 15 nM and miR-34c. This study aimed to investigate the exact inter- vectors (empty vector as NC group) or 35 nM miRNAs (NC miRNA actions between CASC15 and miR-34c in DCRF. as NC group) into 2×10 cells. The following experiments were performed using cells harvested at 24 h after transfection. For all transfections, control (C) cells were untransfected cells. Material and Methods RNA extractions and qPCR Research subjects Total RNAs in 0.3 ml plasma and 2×10 cells were extracted Research subjects of the present study included 50 DCRF pa- using RNAzol (Sigma-Aldrich). To harvest miRNAs, 85% etha- tients (DCRF group, 26 males and 24 females; age 36–67 nol was used to precipitate all RNA samples. All RNA samples years, mean age 51.9±6.0 years), 50 diabetic patients with- were digested with DNase I for 80 min at 37°C to completely out obvious complications (Diabetes group, 26 males and 24 remove genomic DNA. The digested DNA samples were re- females; age 35–68 years, mean age 52.3±6.5 years), and 50 verse-transcribed into cDNA using Tetro Reverse Transcriptase healthy volunteers (Control group, 26 males and 24 females; (Bioline), and cDNA was used as a template to prepare qPCR age 36–68 years, mean age 52.0±6.3 years). All participants mixtures using Brilliant III Ultra-Fast SYBR Green QPCR Master were selected at the Second Affiliated Hospital of Nanchang Mix (Agilent Technologies). With 18S rRNA as an endoge- University between April 2016 and April 2018. All patients were nous control, the expression levels of CASC15 were detected. diagnosed according to the criteria proposed by the Chinese To measure the expression levels of miR-34c, all steps were Medical Association. This study was approved by the hospi- completed using the All-in-OneTM miRNA qRT-PCR Detection tal Ethics Committee. Patient inclusion criteria were: 1) newly Kit (GeneCopoeia). U6 was used as the endogenous control. –DDCT diagnosed cases, 2) no therapies were performed within 100 The 2 method was used to process all data. All PCR reac- days before admission, and 3) CRF caused by diabetes without tions were performed in triplicate. Indexed in: [Current Contents/Clinical Medicine] [SCI Expanded] [ISI Alerting System] This work is licensed under Creative Common Attribution- [ISI Journals Master List] [Index Medicus/MEDLINE] [EMBASE/Excerpta Medica] e919415-2 NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0) [Chemical Abstracts/CAS] Qin X. et al.: LncRNA CASC15 regulates renal cell apoptosis CLINICAL RESEARCH © Med Sci Monit, 2020; 26: e919415 Control (n=50) Control (n=50) Control (n=50) B C 100 100 80 80 60 60 40 40 Sensitivity% Sensitivity% 20 20 Identity% Identity% 0 0 02040 60 80 100 02040 60 80 100 100%-Speci city% 100%-Speci city% Figure 1. Plasma CASC15 was upregulated in DCRF and showed diagnostic values. Levels of CASC15 in plasma derived from 3 groups of participants (DCRF, Diabetes, and Control) were measured by qPCR and compared by ANOVA (one-way) and Tukey test (A). In ROC curve, true-positive cases were DCRF patients and true-negative cases were either diabetic patients (B) or healthy controls (C). Experiments were performed in 3 replicates and mean values are presented, * p<0.05. Cell apoptosis analysis different patient and cell groups by performing ANOVA (one- way) and Tukey test. The diagnostic analysis was performed CIHP-1 cells were harvested at 24 h after transfection and using ROC curves. In ROC curves, true-positive cases were DCRF counted. To prepare a single-cell suspension, 4×10 cells were patients and true-negative cases were either diabetic patients mixed with 1 ml of the cell culture medium. Cells were trans- or healthy controls. p<0.05 was statistically significant. ferred to a 6-well plate with 2 ml cell suspension per well, fol- lowed by the addition of 30 mM D-glucose. Cells were cultivat- ed for 48 h under the aforementioned conditions. After that, Results cells were washed with PBS, and 0.25% trypsin digestion was performed. Finally, cells were stained with Annexin V-FITC and Plasma CASC15 was upregulated in DCRF and showed propidium iodide (PI) for 20 min in the dark, and flow cytome - diagnostic values try was then performed to separate apoptotic cells. Levels of CASC15 in plasma derived from 3 groups of partici- Statistical analysis pants (DCRF, Diabetes, and Control) were measured by qPCR and compared by one-way ANOVA and Tukey test. Comparing The mean values of 3 biological replicates were calculated and to the Control group, significantly higher plasma levels of were used in all data analyses. Differences were explored among CASC15 were observed in the DCRF group but not in the Indexed in: [Current Contents/Clinical Medicine] [SCI Expanded] [ISI Alerting System] This work is licensed under Creative Common Attribution- [ISI Journals Master List] [Index Medicus/MEDLINE] [EMBASE/Excerpta Medica] e919415-3 NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0) [Chemical Abstracts/CAS] 100% Sensitivity Relative CASC15 level 100% Sensitivity Qin X. et al.: LncRNA CASC15 regulates renal cell apoptosis CLINICAL RESEARCH © Med Sci Monit, 2020; 26: e919415 Figure 2. Upregulation of CASC15 was induced by high glucose in CIHP-1 cells. CIHP-1 cells were treated with 5, 10, 20, 5 5 mM 10 mM and 30 mM D-glucose for 12, 24, and 48 h, followed by 20 mM measuring the expression levels of CASC15 by qPCR. 30 mM Data were compared by ANOVA (one-way) and Tukey test. Experiments were performed in 3 replicates and mean values are presented, * p<0.05. 0 12 24 48 * * 5 * 8 * C NC CASC15 C NC CASC15 1.5 1.5 1.0 1.0 0.5 0.5 0.0 0.0 C NC CASC15 C NC CASC15 2 2 0 miR-34c 5’ - AG GG UUGC-3’ GCA UGUAGUUA CUGA CASC15 UGU ACAUCAGU GACU 3’-AAU . . . AAUUG AG G CCCAU . . . AGG-5’ 510 491 Figure 3. CASC15 can sponge miR-34c. CIHP-1 cells were transfected with the CASC15 expression vector and miR-34c mimic. Overexpression of CASC15 and miR-34c was confirmed at 24 h after transfection by qPCR ( A). The interaction between CASC15 and miR-34c was analyzed by qPCR (B). IntaRNA was used to predict the interaction between miR-34c and CASC15 (C). Experiments were performed in 3 replicates and mean values are presented, * p<0.05. Indexed in: [Current Contents/Clinical Medicine] [SCI Expanded] [ISI Alerting System] This work is licensed under Creative Common Attribution- [ISI Journals Master List] [Index Medicus/MEDLINE] [EMBASE/Excerpta Medica] e919415-4 NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0) [Chemical Abstracts/CAS] Relative expression level of CASC15 Relative miR-34c level Relative CASC15 level Relative CASC15 level Relative miR-34c level Qin X. et al.: LncRNA CASC15 regulates renal cell apoptosis CLINICAL RESEARCH © Med Sci Monit, 2020; 26: e919415 C NC miRNA pcDNA3.1 4 4 4 10 10 10 3 3 3 10 10 10 2 2 2 PI 10 PI 10 PI 10 1 1 1 10 10 10 0 0 0 10 10 10 0 1 2 3 4 0 1 2 3 4 0 1 2 3 4 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 miR-34c CASC15 CASC15+miR-34c 4 4 4 10 10 10 3 3 3 10 10 10 2 2 2 PI 10 PI 10 PI 10 1 1 1 10 10 10 0 0 0 10 10 10 0 1 2 3 4 0 1 2 3 4 0 1 2 3 4 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 C NC CASC15 CASC15 CASC15 CASC15 Figure 4. CASC15 attenuated the effects of miR-34c on cell apoptosis. Cell apoptosis assay was performed to analyze the effects of transfections on apoptosis of CIHP-1 cells. Experiments were performed in 3 replicates and mean values are presented, * p<0.05. Diabetes group (Figure 1, p<0.05). The diagnostic value of plas- Upregulation of CASC15 was induced by high glucose in ma CASC15 for DCRF was analyzed by ROC curve. AUC>0.65 CIHP-1 cells indicates potential diagnostic value. With diabetic patients as true-negative cases, the AUC was 0.97 (95% confidence inter - CIHP-1 cells were treated with 5, 10, 20, and 30 mM D-glucose val: 0.95–0.99; standard error: 0.013). With healthy controls as for 12, 24, and 48 h, followed by measuring the expression lev- true-negative cases, the AUC was 0.97 (95% confidence inter - els of CASC15 by qPCR. As shown in Figure 2, expression levels val: 0.95–1.00; standard error: 0.013). AUC>0.65 indicated di- of CASC15 in CIHP-1 cells were upregulated by D-glucose in a agnostic values. Therefore, plasma CASC15 can serve as a di- dose- and time-dependent manner (Figure 2, p<0.05). However, agnostic marker for DCRF. 5 mM D-glucose failed to affect CASC15 expression, possibly because 5 mM is within the normal range. Indexed in: [Current Contents/Clinical Medicine] [SCI Expanded] [ISI Alerting System] This work is licensed under Creative Common Attribution- [ISI Journals Master List] [Index Medicus/MEDLINE] [EMBASE/Excerpta Medica] e919415-5 NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0) [Chemical Abstracts/CAS] Apoptotic cell percentage Qin X. et al.: LncRNA CASC15 regulates renal cell apoptosis CLINICAL RESEARCH © Med Sci Monit, 2020; 26: e919415 CASC15 can sponge miR-34c environment led to significantly upregulated expression of CASC15 in podocytes. The possible reason is that the upregu- CIHP-1 cells were transfected with the CASC15 expression vector lated CASC15 in podocytes fails to affect the plasma levels of and miR-34c mimic. Expression levels of CASC15 and miR-34c CASC15 in diabetic patients. With the development of DCRF, were measured at 24 h after transfection. Compared to the C CASC15 is further upregulated and more CASC15 is released and NC groups, expression levels of CASC15 and miR-34c were into blood, resulting in increased plasma levels of CASC15. significantly increased after transfections (Figure 3A, p<0.05). However, overexpression of CASC15 and miR-34c failed to af- This study showed that the upregulation of plasma CASC15 fect the expression of each other (Figure 3B). Bioinformatics could be used to assist in diagnosis of DCRF, but the accura- analysis using IntaRNA showed that miR-34c can bind CASC15 cy remains to be further analyzed. We showed that CASC15 from position 492 to 509 (Figure 3C). can interact with miR-34c, while overexpression experiments showed that CASC15 and miR-34c have no effects on the ex - CASC15 attenuated the effects of miR-34c on cell pression of each other. Therefore, it appears that miR-34c apoptosis does not target CASC15. It is known that lncRNAs can sponge miRNAs to attenuate their functions [16,17]. The main role of Cell apoptosis assay was performed to analyze the effects of a miRNA sponge is to inhibit the roles of miRNA. This study transfections on apoptosis of CIHP-1 cells. Compared to the revealed that CASC15 attenuated the effects of miR-34c on C and NC groups, overexpression of CASC15 led to increased cell apoptosis. Therefore, CASC15 can sponge miR-34c to pro- apoptotic rate of CIHP-1 cells and miR-34c overexpression led mote the apoptosis of podocytes. to a decreased rate. In addition, CASC15 overexpression at- tenuated the effects of miR-34c overexpression on cell apop - CASC15 is dysregulated in multiple clinical disorders, such as dif- tosis (Figure 4, p<0.05). ferent types of cancers and cardiac hypertrophy [11,15,18–20]. Therefore, the use of CASC15 as a diagnostic marker for DCRF should exclude the possibility of these clinical disorders. In ad- Discussion dition, this study was limited by its small sample size. Studies with larger sample sizes are needed to confirm our conclusions. In this study, we investigated the functions of CASC15 in DCRF and also analyzed the clinical value. We found that CASC15 was upregulated in DCRF and can sponge miR-34c to promote Conclusions the apoptosis of podocytes. In conclusion, CASC15 was upregulated in DCRF and can sponge Previous studies have revealed many lncRNAs involved in the miR-34c to promote the apoptosis of podocytes. Our study pro- development of renal disorders. For instance, lncRNA Erbb4-IR vides novel insights into the molecular mechanism of DCRF can target miR-29b to promote renal injury under a high-glu- and suggests potential therapeutic targets. cose environment [12]. lncRNA MALAT1 was reported to be up- regulated in diabetic patients, and downregulation of MALAT1 Availability of data and materials can promote recovery of renal functions in a rat model [13]. In another study, Wang et al. showed that p53 can downregu- The analyzed datasets generated during the study are avail- late lncRNA ZEB1-AS1 to participate in renal fibrosis in patients able from the corresponding author on reasonable request. with diabetic nephropathy [14]. CASC15 is a well-characterized oncogenic lncRNA [11,15]. CASC15 is upregulated in cancers, Ethics approval and consent to participate and the overexpression of CASC15 promotes cancer progres- sion by regulating cancer cell behaviors, such as inhibition of The present study was approved by the Ethics Committee of cell apoptosis [15]. The present study is the first to report the the Second Alffi iated Hospital of Nanchang University. The re - upregulation of CASC15 and the increased cell apoptosis rate search was carried out in accordance with the World Medical after CASC15 overexpression. Therefore, CASC15 can contrib- Association Declaration of Helsinki. All patients and healthy ute to the development of DCRF. Our study also showed that volunteers provided written informed consent prior to their CASC15 can have opposite roles in regulating cell apoptosis inclusion in the study. in different diseases. Conflict of interests Our study did not observe the upregulation of CASC15 in plas- ma derived from diabetic patients. However, a high-glucose None. Indexed in: [Current Contents/Clinical Medicine] [SCI Expanded] [ISI Alerting System] This work is licensed under Creative Common Attribution- [ISI Journals Master List] [Index Medicus/MEDLINE] [EMBASE/Excerpta Medica] e919415-6 NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0) [Chemical Abstracts/CAS] Qin X. et al.: LncRNA CASC15 regulates renal cell apoptosis CLINICAL RESEARCH © Med Sci Monit, 2020; 26: e919415 References: 1. Zimmet P, Alberti KG, Magliano DJ et al: Diabetes mellitus statistics on prev- 12. Sun SF, Tang PMK, Feng M et al: Novel lncRNA Erbb4-IR promotes diabetic alence and mortality: facts and fallacies. Nat Rev Endocrinol, 2016; 12(10): kidney injury in db/db mice by targeting miR-29b. Diabetes, 2018; 67(4): 616–22 731–44 2. Rowley WR, Bezold C, Arikan Y et al: Diabetes 2030: Insights from yester- 13. Wu D, Cheng Y, Huang X et al: Down-regulation of lncRNA MALAT1 con- day, today, and future trends. Popul Health Manag, 2017; 20(1): 6–12 tributes to renal functional improvement after duodenal-jejunal bypass in a diabetic rat model. J Physiol Biochem, 2018; 74(3): 431–39 3. American Diabetes Association: Economic costs of diabetes in the US in 2017. Diabetes Care, 2018; 41(5): 917–28 14. Wang J, Pang J, Li H et al: lncRNA ZEB1-AS1 was suppressed by p53 for renal fibrosis in diabetic nephropathy. Mol Ther Nucleic Acids, 2018; 12: 741–50 4. Brownlee M: The pathobiology of diabetic complications: A unifying mech- anism. Diabetes, 2005; 54(6): 1615–25 15. Fernando TR, Contreras JR, Zampini M et al: The lncRNA CASC15 regulates SOX4 expression in RUNX1-rearranged acute leukemia. Mol Cancer, 2017; 5. Testa R, Bonfigli A, Genovese S et al: The possible role of flavonoids in the 16(1): 126 prevention of diabetic complications. Nutrients, 2016; 8(5): pii: E310 16. Liang WC, Fu WM, Wong CW et al: The lncRNA H19 promotes epithelial 6. Bos-Touwen I, Schuurmans M, Monninkhof EM et al: Patient and disease to mesenchymal transition by functioning as miRNA sponges in colorectal characteristics associated with activation for self-management in patients cancer. Oncotarget, 2015; 6(26): 22513–25 with diabetes, chronic obstructive pulmonary disease, chronic heart failure and chronic renal disease: A cross-sectional survey study. PLoS One, 2015; 17. Liu D, Li Y, Luo G et al: LncRNA SPRY4-IT1 sponges miR-101-3p to promote 10(5): e0126400 proliferation and metastasis of bladder cancer cells through up-regulating EZH2. Cancer Lett, 2017; 388: 281–91 7. Miranda-Díaz AG, Pazarín-Villaseñor L, Yanowsky-Escatell FG et al: Oxidative stress in diabetic nephropathy with early chronic kidney disease. J Diabetes 18. Yao XM, Tang JH, Zhu H et al: High expression of LncRNA CASC15 is a risk Res, 2016; 2016: 7047238 factor for gastric cancer prognosis and promote the proliferation of gas- tric cancer. Eur Rev Med Pharmacol Sci, 2017; 21(24): 5661–67 8. Ahlqvist E, Van Zuydam NR, Groop LC et al: The genetics of diabetic com- plications. Nat Rev Nephrol, 2015; 11(5): 277–87 19. Li C, Zhou G, Feng J et al: Up-regulation of lncRNA VDR/CASC15 induced by facilitates cardiac hypertrophy through modulating miR-432-5p/TLR4 axis. 9. Ma RCW: Genetics of cardiovascular and renal complications in diabetes. J Biochem Biophys Res Commun, 2018; 503(4): 2407–14 Diabetes Investig, 2016; 7(2): 139–54 20. Jing N, Huang T, Guo H et al: LncRNA CASC15 promotes colon cancer cell pro- 10. Liu XD, Zhang LY, Zhu TC et al: Overexpression of miR-34c inhibits high glu- liferation and metastasis by regulating the miR-4310/LGR5/Wnt/b-catenin cose-induced apoptosis in podocytes by targeting Notch signaling path- signaling pathway. Mol Med Rep, 2018; 18(2): 2269–76 ways. Int J Clin Exp Pathol, 2015; 8(5): 4525–34 11. He T, Zhang L, Kong Y et al: Long non-coding RNA CASC15 is up-regulat- ed in hepatocellular carcinoma and facilitates hepatocarcinogenesis. Int J Oncol, 2017; 51(6): 1722–30 Indexed in: [Current Contents/Clinical Medicine] [SCI Expanded] [ISI Alerting System] This work is licensed under Creative Common Attribution- [ISI Journals Master List] [Index Medicus/MEDLINE] [EMBASE/Excerpta Medica] e919415-7 NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0) [Chemical Abstracts/CAS]

Journal

Medical Science Monitor : International Medical Journal of Experimental and Clinical Research – Pubmed Central

Published: Feb 13, 2020

There are no references for this article.