Similar articles

Abstract: Objective: We aimed to perform a preliminary study of the association between induced pluripotent stem cell (iPS)-related genes and biological behavior of human colorectal cancer (CRC) cells, and the potential for developing anti-cancer drugs targeting these genes. Methods: We used real-time reverse transcriptase polymerase chain reaction (RT-PCR) to evaluate the transcript levels of iPS-related genes NANOG, OCT4, SOX2, C-MYC and KLF4 in CRC cell lines and cancer stem cells (CSCs)-enriched tumor spheres. NANOG was knockdowned in CRC cell line SW620 by lentiviral transduction. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays, plate colony formation, and a mouse xenograft model were used to evaluate alterations in biological behavior in NANOG-knockdown SW620 cells. Also, mock-knockdown and NANOG-knockdown cells were treated with 5-Fluorouracil (5-FU) and survival rate was measured by MTT assay to evaluate drug sensitivity. Results: A significant difference in the transcript levels of iPS-related genes between tumor spheres and their parental bulky cells was observed. NANOG knockdown suppressed proliferation, colony formation, and in vivo tumorigenicity but increased the sensitivity to 5-FU of SW620 cells. 5-FU treatment greatly inhibited the expression of the major stemness-associated genes NANOG, OCT4, and SOX2. Conclusions: These results collectively suggest an overlap between iPS-related genes and CSCs in CRC. Quenching a certain gene NANOG may truncate the aggressiveness of CRC cells.

[1]Almstrup, K., Hoei-Hansen, C.E., Wirkner, U., Blake, J., Schwager, C., Ansorge, W., Nielsen, J.E., Skakkebaek, N.E., Rajpert-de Meyts, E., Leffers, H., 2004. Embryonic stem cell-like features of testicular carcinoma in situ revealed by genome-wide gene expression profiling. Cancer Res., 64(14):4736-4743.

[2]Bae, K.M., Su, Z., Frye, C., McClellan, S., Allan, R.W., Andrejewski, J.T., Kelley, V., Jorgensen, M., Steindler, D.A., Vieweg, J., et al., 2010. Expression of pluripotent stem cell reprogramming factors by prostate tumor initiating cells. J. Urol., 183(5):2045-2053.

[3]Ben-Porath, I., Thomson, M.W., Carey, V.J., Ge, R., Bell, G.W., Regev, A., Weinberg, R.A., 2008. An embryonic stem cell-like gene expression signature in poorly differentiated aggressive human tumors. Nat. Genet., 40(5):499-507.

[4]Bjerkvig, R., Tysnes, B.B., Aboody, K.S., Najbauer, J., Terzis, A.J., 2005. Opinion: the origin of the cancer stem cell: current controversies and new insights. Nat. Rev. Cancer, 5(11):899-904.

[5]Cao, L., Zhou, Y., Zhai, B., Liao, J., Xu, W., Zhang, R., Li, J., Zhang, Y., Chen, L., Qian, H., et al., 2011. Sphere-forming cell subpopulations with cancer stem cell properties in human hepatoma cell lines. BMC Gastroenterol., 11(1):71.

[6]Carette, J.E., Pruszak, J., Varadarajan, M., Blomen, V.A., Gokhale, S., Camargo, F.D., Wernig, M., Jaenisch, R., Brummelkamp, T.R., 2010. Generation of iPSCs from cultured human malignant cells. Blood, 115(20):4039-4042.

[7]Dallas, N.A., Xia, L., Fan, F., Gray, M.J., Gaur, P., van Buren, G.II, Samuel, S., Kim, M.P., Lim, S.J., Ellis, L.M., 2009. Chemoresistant colorectal cancer cells, the cancer stem cell phenotype, and increased sensitivity to insulin-like growth factor-I receptor inhibition. Cancer Res., 69(5):1951-1957.

[8]Dontu, G., Abdallah, W.M., Foley, J.M., Jackson, K.W., Clarke, M.F., Kawamura, M.J., Wicha, M.S., 2003. In vitro propagation and transcriptional profiling of human mammary stem/progenitor cells. Genes Dev., 17(10):1253-1270.

[9]Dylla, S.J., Beviglia, L., Park, I.K., Chartier, C., Raval, J., Ngan, L., Pickell, K., Aguilar, J., Lazetic, S., Smith-Berdan, S., et al., 2008. Colorectal cancer stem cells are enriched in xenogeneic tumors following chemotherapy. PLoS One, 3(6):e2428.

[10]Fan, X., Ouyang, N., Teng, H., Yao, H., 2011. Isolation and characterization of spheroid cells from the HT29 colon cancer cell line. Int. J. Colorectal Dis., 26(10):1279-1285.

[11]Fang, X., Yu, W., Li, L., Shao, J., Zhao, N., Chen, Q., Ye, Z., Lin, S.C., Zheng, S., Lin, B., 2010. ChIP-seq and functional analysis of the SOX2 gene in colorectal cancers. OMICS, 14(4):369-384.

[12]Freyer, J.P., Sutherland, R.M., 1980. Selective dissociation and characterization of cells from different regions of multicell tumor spheroids. Cancer Res., 40(11):3956-3965.

[13]Galli, R., Binda, E., Orfanelli, U., Cipelletti, B., Gritti, A., de Vitis, S., Fiocco, R., Foroni, C., Dimeco, F., Vescovi, A., 2004. Isolation and characterization of tumorigenic, stem-like neural precursors from human glioblastoma. Cancer Res., 64(19):7011-7021.

[14]Jeter, C.R., Badeaux, M., Choy, G., Chandra, D., Patrawala, L., Liu, C., Calhoun-Davis, T., Zaehres, H., Daley, G.Q., Tang, D.G., 2009. Functional evidence that the self-renewal gene NANOG regulates human tumor development. Stem Cells, 27(5):993-1005.

[15]Kelly, P.N., Dakic, A., Adams, J.M., Nutt, S.L., Strasser, A., 2007. Tumor growth need not be driven by rare cancer stem cells. Science, 317(5836):337.

[16]Korkaya, H., Wicha, M.S., 2010. Cancer stem cells: nature versus nurture. Nat. Cell Biol., 12(5):419-421.

[17]Lacerda, L., Pusztai, L., Woodward, W.A., 2010. The role of tumor initiating cells in drug resistance of breast cancer: implications for future therapeutic approaches. Drug Resist. Updat., 13(4-5):99-108.

[18]Lapidot, T., Sirard, C., Vormoor, J., Murdoch, B., Hoang, T., Caceres-Cortes, J., Minden, M., Paterson, B., Caligiuri, M.A., Dick, J.E., 1994. A cell initiating human acute myeloid leukaemia after transplantation into SCID mice. Nature, 367(6464):645-648.

[19]Li, F., Tiede, B., Massague, J., Kang, Y., 2007. Beyond tumorigenesis: cancer stem cells in metastasis. Cell Res., 17(1):3-14.

[20]Ma, S., Tang, K.H., Chan, Y.P., Lee, T.K., Kwan, P.S., Castilho, A., Ng, I., Man, K., Wong, N., To, K.F., et al., 2010. miR-130b Promotes CD133⁺ liver tumor-initiating cell growth and self-renewal via tumor protein 53-induced nuclear protein 1. Cell Stem Cell, 7(6):694-707.

[21]Meng, H.M., Zheng, P., Wang, X.Y., Liu, C., Sui, H.M., Wu, S.J., Zhou, J., Ding, Y.Q., Li, J.M., 2010. Overexpression of Nanog predicts tumor progression and poor prognosis in colorectal cancer. Cancer Biol. Ther., 9(4):295-302.

[22]Miyoshi, N., Ishii, H., Nagai, K., Hoshino, H., Mimori, K., Tanaka, F., Nagano, H., Sekimoto, M., Doki, Y., Mori, M., 2010. Defined factors induce reprogramming of gastrointestinal cancer cells. PNAS, 107(1):40-45.

[23]Morrison, S.J., Kimble, J., 2006. Asymmetric and symmetric stem-cell divisions in development and cancer. Nature, 441(7097):1068-1074.

[24]Park, I.H., Zhao, R., West, J.A., Yabuuchi, A., Huo, H., Ince, T.A., Lerou, P.H., Lensch, M.W., Daley, G.Q., 2008. Reprogramming of human somatic cells to pluripotency with defined factors. Nature, 451(7175):141-146.

[25]Quintana, E., Shackleton, M., Sabel, M.S., Fullen, D.R., Johnson, T.M., Morrison, S.J., 2008. Efficient tumour formation by single human melanoma cells. Nature, 456(7222):593-598.

[26]Reya, T., Morrison, S.J., Clarke, M.F., Weissman, I.L., 2001. Stem cells, cancer, and cancer stem cells. Nature, 414(6859):105-111.

[27]Robertson, F.M., Ogasawara, M.A., Ye, Z., Chu, K., Pickei, R., Debeb, B.G., Woodward, W.A., Hittelman, W.N., Cristofanilli, M., Barsky, S.H., 2010. Imaging and analysis of 3D tumor spheroids enriched for a cancer stem cell phenotype. J. Biomol. Screen., 15(7):820-829.

[28]Saigusa, S., Tanaka, K., Toiyama, Y., Yokoe, T., Okugawa, Y., Ioue, Y., Miki, C., Kusunoki, M., 2009. Correlation of CD133, OCT4, and SOX2 in rectal cancer and their association with distant recurrence after chemoradiotherapy. Ann. Surg. Oncol., 16(12):3488-3498.

[29]Saiki, Y., Ishimaru, S., Mimori, K., Takatsuno, Y., Nagahara, M., Ishii, H., Yamada, K., Mori, M., 2009. Comprehensive analysis of the clinical significance of inducing pluripotent stemness-related gene expression in colorectal cancer cells. Ann. Surg. Oncol., 16(9):2638-2644.

[30]Shackleton, M., Quintana, E., Fearon, E.R., Morrison, S.J., 2009. Heterogeneity in cancer: cancer stem cells versus clonal evolution. Cell, 138(5):822-829.

[31]Sherar, M.D., Noss, M.B., Foster, F.S., 1987. Ultrasound backscatter microscopy images the internal structure of living tumour spheroids. Nature, 330(6147):493-495.

[32]Singh, A., Settleman, J., 2010. EMT, cancer stem cells and drug resistance: an emerging axis of evil in the war on cancer. Oncogene, 29(34):4741-4751.

[33]Singh, S.K., Hawkins, C., Clarke, I.D., Squire, J.A., Bayani, J., Hide, T., Henkelman, R.M., Cusimano, M.D., Dirks, P.B., 2004. Identification of human brain tumour initiating cells. Nature, 432(7015):396-401.

[34]Takahashi, K., Tanabe, K., Ohnuki, M., Narita, M., Ichisaka, T., Tomoda, K., Yamanaka, S., 2007. Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell, 131(5):861-872.

[35]Tang, C., Ang, B.T., Pervaiz, S., 2007. Cancer stem cell: target for anti-cancer therapy. FASEB J., 21(14):3777-3785.

[36]Tindall, M.J., Please, C.P., 2007. Modelling the cell cycle and cell movement in multicellular tumour spheroids. Bull. Math. Biol., 69(4):1147-1165.

[37]Todaro, M., Alea, M.P., di Stefano, A.B., Cammareri, P., Vermeulen, L., Iovino, F., Tripodo, C., Russo, A., Gulotta, G., Medema, J.P., et al., 2007. Colon cancer stem cells dictate tumor growth and resist cell death by production of interleukin-4. Cell Stem Cell, 1(4):389-402.

[38]Waleh, N.S., Brody, M.D., Knapp, M.A., Mendonca, H.L., Lord, E.M., Koch, C.J., Laderoute, K.R., Sutherland, R.M., 1995. Mapping of the vascular endothelial growth factor-producing hypoxic cells in multicellular tumor spheroids using a hypoxia-specific marker. Cancer Res., 55(24):6222-6226.

[39]Wang, X., Kruithof-de Julio, M., Economides, K.D., Walker, D., Yu, H., Halili, M.V., Hu, Y.P., Price, S.M., Abate-Shen, C., Shen, M.M., 2009. A luminal epithelial stem cell that is a cell of origin for prostate cancer. Nature, 461(7263):495-500.

[40]Wei, D., Kanai, M., Huang, S., Xie, K., 2006. Emerging role of KLF4 in human gastrointestinal cancer. Carcinogenesis, 27(1):23-31.

[41]Wibe, E., 1980. Resistance to vincristine of human cells grown as multicellular spheroids. Br. J. Cancer, 42(6):937-941.

[42]Yeung, T.M., Gandhi, S.C., Wilding, J.L., Muschel, R., Bodmer, W.F., 2010. Cancer stem cells from colorectal cancer-derived cell lines. PNAS, 107(8):3722-3727.

[43]Yin, B.B., Wu, S.J., Zong, H.J., Ma, B.J., Cai, D., 2011. Preliminary screening and identification of stem cell-like sphere clones in a gallbladder cancer cell line GBC-SD. J. Zhejiang Univ.-Sci. B (Biomed. & Biotechnol.), 12(4):256-263.

[44]Yori, J.L., Johnson, E., Zhou, G., Jain, M.K., Keri, R.A., 2010. Kruppel-like factor 4 inhibits epithelial-to-mesenchymal transition through regulation of E-cadherin gene expression. J. Biol. Chem., 285(22):16854-16863.

[45]Yu, J., Vodyanik, M.A., Smuga-Otto, K., Antosiewicz-Bourget, J., Frane, J.L., Tian, S., Nie, J., Jonsdottir, G.A., Ruotti, V., Stewart, R., et al., 2007. Induced pluripotent stem cell lines derived from human somatic cells. Science, 318(5858):1917-1920.

[46]Zhao, W., Hisamuddin, I.M., Nandan, M.O., Babbin, B.A., Lamb, N.E., Yang, V.W., 2004. Identification of Kruppel-like factor 4 as a potential tumor suppressor gene in colorectal cancer. Oncogene, 23(2):395-402.