shRNA targeting Bmi1 impedes the self-renewal of cisplatin-enriched stem-like cells in human A549 cells

Xu,Yu; Hu,Yi-De; Zhou,Jing; Zhang,Ming-Hui; Yuan,Wei-Wei; Luo,Yan

doi:10.3892/or.2012.1843

shRNA targeting Bmi1 impedes the self-renewal of cisplatin-enriched stem-like cells in human A549 cells

Authors:
- Yu Xu
- Yi-De Hu
- Jing Zhou
- Ming-Hui Zhang
- Wei-Wei Yuan
- Yan Luo
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Affiliations: The Third Department of Oncology, PLA Cancer Research Institute of the Second Affiliated Hospital, The Third Military Medical University, Chongqing 400037, P.R. China
Published online on: June 1, 2012 https://doi.org/10.3892/or.2012.1843
Pages: 629-639

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Abstract

It has been hypothesized that cancer stem-like cells are responsible for tumor recurrence following chemotherapy. Evidence on the mechanisms through which drug-resistant stem-like cells recapitulate the tumor mass has not been definitively reported. Based on this information, we investigated the enrichment ability of a population of stem-like cells following treatment with cisplatin in human A549 cells and focused on the molecular mechanisms regulating the self-renewal of stem-like cells. A population of stem-like cells was enriched following cisplatin treatment and was defined phenotypically and functionally based on the expression of certain stem cell markers, sphere-forming ability, multipotent differentiation and induction of xenograft tumors in vivo. For various types of differentiated cells, Bmi1 has been reported to be important for cell proliferation and for the self-renewal of stem cells. The high expression of Bmi1 in cisplatin-enriched stem-like cells was shown using Q-PCR and western blotting; therefore, the role of Bmi1 was investigated in cisplatin-enriched stem-like cells by infecting cisplatin-enriched stem-like cells with Bmi1-targeted RNAi lentiviruses. Cell proliferation, tumor sphere formation and xenograft formation was reduced following knockdown of Bmi1. Based on our results, we propose that, after cisplatin treatment, Bmi1 is required for the self-renewal of stem-like cells that are important for the expansion of the stem-like cell pool in human A549 cells and that targeting Bmi1 slows down the formation of tumors in vivo.

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1	Shackleton M, Quintana E, Fearon ER and Morrison SJ: Heterogeneity in cancer: cancer stem cells versus clonal evolution. Cell. 138:822–829. 2009. View Article : Google Scholar : PubMed/NCBI
2	Vermeulen L, Todaro M, de Sousa Mello F, et al: Single-cell cloning of colon cancer stem cells reveals a multi-lineage differentiation capacity. Proc Natl Acad Sci USA. 105:13427–13432. 2008. View Article : Google Scholar : PubMed/NCBI
3	Dick JE: Stem cell concepts renew cancer research. Blood. 112:4793–4807. 2008. View Article : Google Scholar : PubMed/NCBI
4	Bonnet D and Dick JE: Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hematopoietic cell. Nat Med. 3:730–737. 1997. View Article : Google Scholar : PubMed/NCBI
5	Harrison H, Farnie G, Howell SJ, et al: Regulation of breast cancer stem cell activity by signaling through the Notch4 receptor. Cancer Res. 70:709–718. 2010. View Article : Google Scholar : PubMed/NCBI
6	Visvader JE and Lindeman GJ: Cancer stem cells in solid tumours: accumulating evidence and unresolved questions. Nat Rev Cancer. 8:755–768. 2008. View Article : Google Scholar : PubMed/NCBI
7	Kim CF, Jackson EL, Woolfenden AE, et al: Identification of bronchioalveolar stem cells in normal lung and lung cancer. Cell. 121:823–835. 2005. View Article : Google Scholar : PubMed/NCBI
8	Eramo A, Lotti F, Sette G, et al: Identification and expansion of the tumorigenic lung cancer stem cell population. Cell Death Differ. 15:504–514. 2008. View Article : Google Scholar : PubMed/NCBI
9	Seo DC, Sung JM, Cho HJ, et al: Gene expression profiling of cancer stem cell in human lung adenocarcinoma A549 cells. Mol Cancer. 6:752007. View Article : Google Scholar : PubMed/NCBI
10	Ho MM, Ng AV, Lam S and Hung JY: Side population in human lung cancer cell lines and tumors is enriched with stem-like cancer cells. Cancer Res. 67:4827–4833. 2007. View Article : Google Scholar : PubMed/NCBI
11	Al-Hajj M, Wicha MS, Benito-Hernandez A, Morrison SJ and Clarke MF: Prospective identification of tumorigenic breast cancer cells. Proc Natl Acad Sci USA. 100:3983–3988. 2003. View Article : Google Scholar : PubMed/NCBI
12	Sullivan JP, Minna JD and Shay JW: Evidence for self-renewing lung cancer stem cells and their implications in tumor initiation, progression, and targeted therapy. Cancer Metastasis Rev. 29:61–72. 2010. View Article : Google Scholar : PubMed/NCBI
13	Maeda R, Yoshida J, Hishida T, et al: Late recurrence of non-small cell lung cancer more than 5 years after complete resection: incidence and clinical implications in patient follow-up. Chest. 138:145–150. 2010.PubMed/NCBI
14	Dean M, Fojo T and Bates S: Tumour stem cells and drug resistance. Nat Rev Cancer. 5:275–284. 2005. View Article : Google Scholar
15	Dylla SJ, Beviglia L, Park IK, et al: Colorectal cancer stem cells are enriched in xenogeneic tumors following chemotherapy. PLoS One. 3:e24282008. View Article : Google Scholar : PubMed/NCBI
16	Eramo A, Ricci-Vitiani L, Zeuner A, et al: Chemotherapy resistance of glioblastoma stem cells. Cell Death Differ. 13:1238–1241. 2006. View Article : Google Scholar : PubMed/NCBI
17	Tamase A, Muraguchi T, Naka K, et al: Identification of tumor-initiating cells in a highly aggressive brain tumor using promoter activity of nucleostemin. Proc Natl Acad Sci USA. 106:17163–17168. 2009. View Article : Google Scholar : PubMed/NCBI
18	Malanchi I, Peinado H, Kassen D, et al: Cutaneous cancer stem cell maintenance is dependent on beta-catenin signalling. Nature. 452:650–653. 2008. View Article : Google Scholar : PubMed/NCBI
19	Korkaya H, Paulson A, Iovino F and Wicha MS: HER2 regulates the mammary stem/progenitor cell population driving tumorigenesis and invasion. Oncogene. 27:6120–6130. 2008. View Article : Google Scholar : PubMed/NCBI
20	Dovey JS, Zacharek SJ, Kim CF and Lees JA: Bmi1 is critical for lung tumorigenesis and bronchioalveolar stem cell expansion. Proc Natl Acad Sci USA. 105:11857–11862. 2008. View Article : Google Scholar : PubMed/NCBI
21	Levina V, Marrangoni AM, DeMarco R, Gorelik E and Lokshin AE: Drug-selected human lung cancer stem cells: cytokine network, tumorigenic and metastatic properties. PLoS One. 3:e30772008. View Article : Google Scholar : PubMed/NCBI
22	Bea S, Tort F, Pinyol M, et al: BMI-1 gene amplification and overexpression in hematological malignancies occur mainly in mantle cell lymphomas. Cancer Res. 61:2409–2412. 2001.
23	Tsunoda S, Okumura T, Ito T, et al: ABCG2 expression is an independent unfavorable prognostic factor in esophageal squamous cell carcinoma. Oncology. 71:251–258. 2006. View Article : Google Scholar : PubMed/NCBI
24	Donnenberg VS and Donnenberg AD: Multiple drug resistance in cancer revisited: the cancer stem cell hypothesis. J Clin Pharmacol. 45:872–877. 2005. View Article : Google Scholar : PubMed/NCBI
25	Tredan O, Galmarini CM, Patel K and Tannock IF: Drug resistance and the solid tumor microenvironment. J Natl Cancer Inst. 99:1441–1454. 2007. View Article : Google Scholar : PubMed/NCBI
26	Eyler CE and Rich JN: Survival of the fittest: cancer stem cells in therapeutic resistance and angiogenesis. J Clin Oncol. 26:2839–2845. 2008. View Article : Google Scholar : PubMed/NCBI
27	Kanaji N, Bandoh S, Fujita J, Ishii T, Ishida T and Kubo A: Compensation of type I and type II cytokeratin pools in lung cancer. Lung Cancer. 55:295–302. 2007. View Article : Google Scholar : PubMed/NCBI
28	Li L and Neaves WB: Normal stem cells and cancer stem cells: the niche matters. Cancer Res. 66:4553–4557. 2006. View Article : Google Scholar : PubMed/NCBI
29	Beachy PA, Karhadkar SS and Berman DM: Tissue repair and stem cell renewal in carcinogenesis. Nature. 432:324–331. 2004. View Article : Google Scholar : PubMed/NCBI
30	Park IK, Morrison SJ and Clarke MF: Bmi1, stem cells, and senescence regulation. J Clin Invest. 113:175–179. 2004. View Article : Google Scholar : PubMed/NCBI
31	Zhang HW, Ding J, Jin JL, et al: Defects in mesenchymal stem cell self-renewal and cell fate determination lead to an osteopenic phenotype in Bmi-1 null mice. J Bone Miner Res. 25:640–652. 2010. View Article : Google Scholar : PubMed/NCBI
32	Shakhova O, Leung C and Marino S: Bmi1 in development and tumorigenesis of the central nervous system. J Mol Med. 83:596–600. 2005. View Article : Google Scholar : PubMed/NCBI
33	van der Lugt NM, Domen J, Linders K, et al: Posterior transformation, neurological abnormalities, and severe hematopoietic defects in mice with a targeted deletion of the bmi-1 proto-oncogene. Genes Dev. 8:757–769. 1994.