Biochemical and Biophysical Research Communications 377 (2008) 668–673
0006-291X/$ - see front matter © 2008 Elsevier Inc. All rights reserved.
doi:10.1016/j.bbrc.2008.10.052
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Biochemical and Biophysical Research Communications
journal homepage: www.elsevier.com/locate/ybbrc
Can cer stem cells are a rare pop u la tion of undif fer en ti ated
tumor i genic cells respon si ble for tumor ini ti a tion, main te nance
and spread ing. There remains con tro versy as to whether they are
derived from nor mal stem cells or rep re sent mutated mature cells.
How ever, increas ingly the evi dence sug gests that self-nor mal stem
cells are more likely to be the pro gen i tors of lung can cer stem cells.
Pre vi ous injury mod els have iden ti fied sev eral stem cell com part-
ments in the nor mal lung with pro lif er a tive and dif fer en ti a tion
potential. The stem cells resid ing in the bron cho al ve o lar junc-
tion of adult lungs with regen er a tive potential have been fur ther
char ac ter ized. Recently, Kim and col leagues iden ti fied bron chi oal-
ve o lar stem cells (BAS Cs) in nor mal lung and lung can cer. Impor-
tantly, their find ings point to BAS Cs as the puta tive cells of ori gin
for ade no car ci noma [1]. How ever, the under ly ing mech a nisms for
main tain ing the self-renewal capac ity of BAS Cs remain largely
unknown.
Given the pro posed involve ment of BAS Cs in the devel op ment
of ade no car ci noma, elu ci da tion of the reg u la tory mech a nisms
con trol ling their self-renewal, pro lif er a tion and dif fer en ti a tion
could pro vide new ave nues to explore the under ly ing mech a-
nisms of can cer devel op ment. miR NAs are par tic u larly attrac-
tive can di dates for hav ing a role in the main te nance of BASC as
they have cru cial roles in both the devel op ment of nor mal stem
cells and can cer path o gen e sis [2–4]. There fore we devel oped a
reli able, in-house miR NA micro array plat form to inves ti gate the
global micr oR NA expres sion pro file of BASC. Expres sion pro fil ing
of miR NAs in human and mouse organs has been reported by some
groups, but to our knowl edge, this is the first study to inves ti gate
the miR NA expres sion pro file of mouse BAS Cs.
Mate ri als and meth ods
Ani mals. Male C57BL/6J (H-2b) mice, 4 weeks of age, were
obtained from Insti tute of Ani mal, Third Mil i tary Med i cal Uni ver-
sity (Chon gq ing, China). Ani mals were housed under con trolled
tem per a ture, humid ity, and a 12-h light:dark cycle with food and
water ad libi tum. The insti tu tional Ani mal Care and Use Com mit-
tee approved all ani mal pro to cols.
Immu no stain ing on tis sue sec tions and cells from mouse lung. Mice
were killed at the times indi cated and sub jected to full nec ropsy;
tra cheas were removed from the appro pri ate ani mals, fixed for
2 h in 2% para for mal de hyde in PBS on ice, and then equil i brated
over night in 18% sucrose in PBS at 4 °C. Immu no flu o res cent stain-
ing was then per formed accord ing to our pre vi ously pub lished
paper. Rab bit poly clonal anti-CCA at 1:1000, and goat poly clonal
anti-pro-SP-C (Santa Cruz Bio tech nol ogy, Santa Cruz, CA) at 1:50,
dilu tion were used in the pres ent study.
BAS Cs sort ing by flow cytom e try. Mice were sac ri ficed, and their
lungs were removed and cut into small pieces. Lung spec i mens
were washed sev eral times with DMEM–F12 medium. Tis sue
dis so ci a tion was per formed by treat ing with 0.1% pro te ase type-XIV
(Sigma) and
Dis pase in DMEM–F12 (Sigma) at 4 °C over night. After-
wards, tis sues were trans ferred to 10% FCS_Jok lik’s MEM, pipet ted
sev eral times to release pulmonary cells, and then fil tered through
a 100
l
m nylon cell strainer. After wash ing, cells were stained with
PE-con ju gated anti-Sca-1, FITC-con ju gated anti-CD34, PE-CY5-con-
ju gated anti-CD45 and PE-CY7-con ju gated anti-CD31 (eBio science,
MicroRNA expression profile of bronchioalveolar stem cells from mouse lung
Shen Qian, Jin-yong Ding, Rongkai Xie, Jiang-hong An, Xu-jun Ao, Zhen-guo Zhao, Jian-guo Sun,
Yu-zhong Duan, Zheng-tang Chen
*
, Bo Zhu
*
Insti tute of Can cer, Xinqiao Hos pi tal, Third Mil i tary Med i cal Uni ver sity, Xin quiao Street No. 2, Shap ing ba Di, Chon gq ing 400037, PR China
article info abstract
Article history:
Received 2 October 2008
Available online 21 October 2008
Increas ing evi dence has sug gested that bron chi oal ve o lar stem cell (BASC) is the pro gen i tor cells of lung
can cer stem cells. How ever, the mech a nisms by which self-renewal of BSACs is con trolled and how BAS-
Cs turn into can cer stem cells still remains to be unknown. In the pres ent study, we suc cess fully iso lated
bron chi oal ve o lar stem cells (BAS Cs) from mouse lung using FACS. These BAS Cs were char ac ter ized by clonal
growth, self-renewal and high capac ity for dif fer en ti a tion, sug gest ing that these BAS Cs are indeed stem cells.
We inves ti gated the micr oR NA (miR NA) expres sion pro file of these BAS Cs using miR NA array and quan ti ta-
tive RT-PCR. We dis cov ered that BAS Cs pos sessed a unique miR NA pro file, with altered expres sion of sev eral
mi croR NAs, such as miR-142-3p, miR-451, miR-106a, miR-142-5p, miR-15b, miR-20a, miR-106b, miR-25, miR-
486, in BAS Cs com pared to con trol cells. Our results sug gest that mi croR NAs might play impor tant roles in
main tain ing the self-renewal capac ity of BAS Cs, and sug gest the intrigu ing pos si bil ity that aber rant expres-
sion of mi croR NAs could involved in turn ing BAS Cs into lung can cer stem cells.
© 2008 Elsevier Inc. All rights reserved.
Key words:
Bron chi oal ve o lar stem cell
Micr oR NA
Micro array
Lung can cer
*
Cor re spond ing authors.
E-mail addresses: zheng tang chen@yahoo.com.cn (Z. Chen), oncol ogy_bo zhu@
yahoo.com.cn (B. Zhu)