miR-27a regulates the self renewal of the H446 small cell lung cancer cell line in vitro

Miao,Yajing; Li,Jiayin; Qiu,Xiaofei; Li,Yanlei; Wang,Zhengyan; Luan,Yajing

doi:10.3892/or.2012.2095

miR-27a regulates the self renewal of the H446 small cell lung cancer cell line in vitro

Authors:
- Yajing Miao
- Jiayin Li
- Xiaofei Qiu
- Yanlei Li
- Zhengyan Wang
- Yajing Luan
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Affiliations: Department of Pathology, Tianjin Medical University, Tianjin 300070, P.R. China, Liaocheng People's Hospital, Liaocheng 252000, Shandong, P.R. China
Published online on: October 19, 2012 https://doi.org/10.3892/or.2012.2095
Pages: 161-168

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Abstract

Cancer growth is driven by cancer stem-like cells within a tumor, called cancer stem cells (CSCs). Since miRNAs can regulate cell-fate decisions, we compared miRNA expression in stem-like cells and differentiated cells from small cell lung cancer (SCLC) cell lines to develop further understanding of the molecular mechanisms involved in the pathogenesis of SCLC. First, SCLC stem-like cells were enriched by isolating sphere-forming cells using a deﬁned serum-free medium. Further, microRNA microarrays were used to measure the expression of 1212 miRNAs in sphere-forming cells and parental cells. We found 86 miRNAs that were differentially expressed, including 48 upregulated miRNAs and 38 downregulated miRNAs between sphere-forming cells and parental cells. Among them, five downregulated miRNAs (let-7, miR-20, 21, 27a and 30b) and one upregulated miRNA (miR-149*) were selected for validation in 3 sets of SCLC cell lines by qRT-RCR. The qRT-PCR analysis confirmed that all six miRNAs were indeed differentially expressed. However, only miR-27a was consistently downregulated in sphere-forming cells of all 3 cell lines. Antagonizing miR-27a by inhibitor in parental cells enhanced proliferation, self renewal, and the proportion of undifferentiated cells in vitro. The candidate miRNA and some miRNAs with same seed sequence are predicted to have several target genes related to apoptosis, cell proliferation and cell cycle. Our results suggest that downregulation of miR-27a enhanced the stem-like properties of SCLC cells in vitro and may be critical to maintaining a stem cell function in SCLC.

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1	Jackman DM and Johnson BE: Small-cell lung cancer. Lancet. 366:1385–1396. 2005. View Article : Google Scholar : PubMed/NCBI
2	Jemal A, Siegel R, Ward E, Hao Y, Xu J and Thun MJ: Cancer statistics, 2009. CA Cancer J Clin. 59:225–249. 2009. View Article : Google Scholar
3	Govindan R, Page N, Morgensztern D, et al: Changing epidemiology of small-cell lung cancer in the United States over the last 30 years: analysis of the surveillance, epidemiologic, and end results database. J Clin Oncol. 24:4539–4544. 2006.PubMed/NCBI
4	Lally BE, Urbanic JJ, Blackstock AW, Miller AA and Perry MC: Small cell lung cancer: have we made any progress over the last 25 years? Oncologist. 12:1096–1104. 2007.PubMed/NCBI
5	Filipowicz W, Bhattacharyya SN and Sonenberg N: Mechanisms of post-transcriptional regulation by microRNAs: are the answers in sight? Nat Rev Genet. 9:102–114. 2008. View Article : Google Scholar : PubMed/NCBI
6	Shimono Y, Zabala M, Cho RW, et al: Downregulation of miRNA-200c links breast cancer stem cells with normal stem cells. Cell. 138:592–603. 2009. View Article : Google Scholar : PubMed/NCBI
7	Garzon R, Calin GA and Croce CM: MicroRNAs in cancer. Annu Rev Med. 60:167–179. 2009. View Article : Google Scholar
8	Yu F, Yao H, Zhu P, et al: let-7 regulates self renewal and tumorigenicity of breast cancer cells. Cell. 131:1109–1123. 2007. View Article : Google Scholar : PubMed/NCBI
9	Ma S, Tang KH, Chan YP, et al: miR-130b promotes CD133(+) liver tumor-initiating cell growth and self-renewal via tumor protein 53-induced nuclear protein 1. Cell Stem Cell. 7:694–707. 2010.PubMed/NCBI
10	Liu C, Kelnar K, Liu B, et al: The microRNA miR-34a inhibits prostate cancer stem cells and metastasis by directly repressing CD44. Nat Med. 17:211–215. 2011. View Article : Google Scholar : PubMed/NCBI
11	Hua S, Xiaotao X, Renhua G, et al: Reduced miR-31 and let-7 maintain the balance between differentiation and quiescence in lung cancer stem-like side population cells. Biomed Pharmacother. 66:89–97. 2012. View Article : Google Scholar : PubMed/NCBI
12	Ji Q, Hao X, Zhang M, et al: MicroRNA miR-34 inhibits human pancreatic cancer tumor-initiating cells. PLoS One. 4:e68162009. View Article : Google Scholar : PubMed/NCBI
13	Gal H, Pandi G, Kanner AA, et al: MIR-451 and Imatinib mesylate inhibit tumor growth of glioblastoma stem cells. Biochem Biophys Res Commun. 376:86–90. 2008. View Article : Google Scholar : PubMed/NCBI
14	Qiu X, Wang Z, Li Y, Miao Y, Ren Y and Luan Y: Characterization of sphere-forming cells with stem-like properties from the small cell lung cancer cell line H446. Cancer Lett. 323:161–170. 2012. View Article : Google Scholar : PubMed/NCBI
15	Bolstad BM, Irizarry RA, Astrand M and Speed TP: A comparison of normalization methods for high density oligonucleotide array data based on variance and bias. Bioinformatics. 19:185–193. 2003. View Article : Google Scholar : PubMed/NCBI
16	Chen C, Ridzon DA, Broomer AJ, et al: Real-time quantification of microRNAs by stem-loop RT-PCR. Nucleic Acids Res. 33:e1792005. View Article : Google Scholar : PubMed/NCBI
17	Melton C, Judson RL and Blelloch R: Opposing microRNA families regulate self-renewal in mouse embryonic stem cells. Nature. 463:621–626. 2010. View Article : Google Scholar : PubMed/NCBI
18	Liu C and Tang DG: MicroRNA regulation of cancer stem cells. Cancer Res. 71:5950–5954. 2011. View Article : Google Scholar : PubMed/NCBI
19	DeSano JT and Xu L: MicroRNA regulation of cancer stem cells and therapeutic implications. AAPS J. 11:682–692. 2009. View Article : Google Scholar : PubMed/NCBI
20	Zimmerman AL and Wu S: MicroRNAs, cancer and cancer stem cells. Cancer Lett. 300:10–19. 2011. View Article : Google Scholar : PubMed/NCBI
21	Kohu K, Feng J, Iwama A and Satake M: MicroRNA-27 enhances differentiation of myeloblasts into granulocytes by post-transcriptionally downregulating Runx1. Br J Haematol. 145:412–423. 2009. View Article : Google Scholar : PubMed/NCBI
22	Buckingham M, Crist CG, Montarras D, et al: Muscle stem cell behavior is modified by microRNA-27 regulation of Pax3 expression. Proc Natl Acad Sci USA. 106:13383–13387. 2009. View Article : Google Scholar : PubMed/NCBI
23	Takamizawa J, Konishi H, Yanagisawa K, et al: Reduced expression of the let-7 microRNAs in human lung cancers in association with shortened postoperative survival. Cancer Res. 64:3753–3756. 2004. View Article : Google Scholar : PubMed/NCBI
24	Huang GL, Zhang XH, Guo GL, et al: Clinical significance of miR-21 expression in breast cancer: SYBR-Green I-based real-time RT-PCR study of invasive ductal carcinoma. Oncol Rep. 21:673–679. 2009.PubMed/NCBI
25	Shao JY, Yan LX, Huang XF, et al: MicroRNA miR-21 overexpression in human breast cancer is associated with advanced clinical stage, lymph node metastasis and patient poor prognosis. RNA. 14:2348–2360. 2008. View Article : Google Scholar : PubMed/NCBI
26	Asangani IA, Rasheed SA, Nikolova DA, et al: MicroRNA-21 (miR-21) post-transcriptionally downregulates tumor suppressor Pdcd4 and stimulates invasion, intravasation and metastasis in colorectal cancer. Oncogene. 27:2128–2136. 2008. View Article : Google Scholar : PubMed/NCBI
27	Zhang BG, Li JF, Yu BQ, Zhu ZG, Liu BY and Yan M: microRNA-21 promotes tumor proliferation and invasion in gastric cancer by targeting PTEN. Oncol Rep. 27:1019–1026. 2012.PubMed/NCBI
28	Gao W, Shen H, Liu L, Xu J and Shu Y: MiR-21 overexpression in human primary squamous cell lung carcinoma is associated with poor patient prognosis. J Cancer Res Clin Oncol. 137:557–566. 2011. View Article : Google Scholar : PubMed/NCBI
29	Zhu S, Wu H, Wu F, Nie D, Sheng S and Mo YY: MicroRNA-21 targets tumor suppressor genes in invasion and metastasis. Cell Res. 18:350–359. 2008. View Article : Google Scholar : PubMed/NCBI
30	Hiyoshi Y, Kamohara H, Karashima R, et al: MicroRNA-21 regulates the proliferation and invasion in esophageal squamous cell carcinoma. Clin Cancer Res. 15:1915–1922. 2009. View Article : Google Scholar : PubMed/NCBI
31	Horiuchi A, Iinuma H, Akahane T, Shimada R and Watanabe T: Prognostic significance of PDCD4 expression and association with microRNA-21 in each Dukes’ stage of colorectal cancer patients. Oncol Rep. 27:1384–1392. 2012.PubMed/NCBI
32	Zhu Q, Wang Z, Hu Y, et al: miR-21 promotes migration and invasion by the miR-21-PDCD4-AP-1 feedback loop in human hepatocellular carcinoma. Oncol Rep. 27:1660–1668. 2012.PubMed/NCBI
33	Singh SK, Kagalwala MN, Parker-Thornburg J, Adams H and Majumder S: REST maintains self-renewal and pluripotency of embryonic stem cells. Nature. 453:223–227. 2008. View Article : Google Scholar : PubMed/NCBI
34	Houbaviy HB, Murray MF and Sharp PA: Embryonic stem cell-specific MicroRNAs. Dev Cell. 5:351–358. 2003. View Article : Google Scholar : PubMed/NCBI