Similar articles

Abstract: Objective: Side population (SP) cells may play a crucial role in tumorigenesis and the recurrence of cancer. Many kinds of cell lines and tissues have demonstrated the presence of SP cells, including several gastric cancer cell lines. This study is aimed to identify the cancer stem-like cells in the SP of gastric cancer cell line MKN-45. Methods: We used fluorescence activated cell sorting (FACS) to sort SP cells in the human gastric carcinoma cell line MKN-45 (cells labeled with Hoechst 33342) and then characterized the cancer stem-like properties of SP cells. Results: This study found that the SP cells had higher clone formation efficiency than major population (MP) cells. Five stemness-related gene expression profiles, including OCT-4, SOX-2, NANOG, CD44, and adenosine triphosphate (ATP)-binding cassette transporters gene ABCG2, were tested in SP and MP cells using quantitative real-time reverse transcription polymerase chain reaction (RT-PCR). Western blot was used to show the difference of protein expression between SP and MP cells. Both results show that there was significantly higher protein expression in SP cells than in MP cells. When inoculated into non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice, SP cells show higher tumorigenesis tendency than MP cells. Conclusions: These results indicate that SP cells possess cancer stem cell properties and prove that SP cells from MKN-45 are gastric cancer stem-like cells.

[1]Al-Hajj, M., Wicha, M.S., Benito-Hernandez, A., Morrison, S.J., Clarke, M.F., 2003. Prospective identification of tumorigenic breast cancer cells. PNAS, 100(7):3983-3988.

[2]Alvi, A.J., Clayton, H., Joshi, C., Enver, T., Ashworth, A., Vivanco, M.M., Dale, T.C., Smalley, M.J., 2003. Functional and molecular characterization of mammary side population cells. Breast Cancer Res., 5(1):R1-R8.

[3]Boiani, M., Scholer, H.R., 2005. Regulatory networks in embryoderived pluripotent stem cells. Nat. Rev. Mol. Cell Biol., 6(11):872-884.

[4]Boyer, L.A., Lee, T.I., Cole, M.F., Johnstone, S.E., Levie, S.S., Zucker, J.P., Guenther, M.G., Kumar, R.M., Murray, H.L., Jenner, R.G., et al., 2005. Core transcriptional regulatory circuitry in human embryonic stem cells. Cell, 122(6):947-956.

[5]Chambers, I., Colby, D., Robertson, M., Nichols, J., Lee, S., Tweedie, S., Smith, A., 2003. Functional expression cloning of NANOG, a pluripotency sustaining factor in embryonic stem cells. Cell, 113(5):643-655.

[6]Chiba, T., Kita, K., Zheng, Y.W., Yokosuka, O., Saisho, H., Iwama, A., Nakauchi, H., Taniguchi, H., 2006. Side population purified from hepatocellular carcinoma cells harbors cancer stem cell-like properties. Hepatology, 44(1):240-251.

[7]Fukuda, K., Saikawa, Y., Ohashi, M., Kumagai, K., Kitajima, M., Okano, H., Matsuzaki, Y., Kitagawa, Y., 2009. Tumor initiating potential of side population cells in human gastric cancer. Int. J. Oncol., 34(5):1201-1207.

[8]Goodell, M.A., Browse, K., Paradis, G., Conner, A.S., Mulligan, R.C., 1996. Isolation and functional properties of murine hematopoietic stem cells that are replicating in vivo. J. Exp. Med., 183(4):1797-1806.

[9]Hadnagy, A., Gaboury, L., Beaulieu, R., Balicki, D., 2006. SP analysis may be used to identify cancer stem cell populations. Exp. Cell Res., 312(19):3701-3710.

[10]Haraguchi, N., Utsunomiya, T., Inoue, H., Tanaka, F., Mimori, K., Barnard, G.F., Mori, M., 2006. Characterization of a side population of cancer cells from human gastrointestinal system. Stem Cells, 24(3):506-513.

[11]Ho, M.M., Ng, A.V., Lam, S., Hung, J.Y., 2007. Side population in human lung cancer cell lines and tumors is enriched with stem-like cancer cells. Cancer Res., 67(10):4827-4833.

[12]Houghton, J., Stoicov, C., Nomura, S., Rogers, A.B., Carlson, J., Li, H., Cai, X., Fox, J.G., Goldenring, J.R., Wang, T.C., 2004. Gastric cancer originating from bone marrow derived cells. Science, 306(5701):1568-1571.

[13]Huang, D.Z., Gao, Q.L., Guo, L.P., Zhang, C.P., Jiang, W., Li, H.X., Wang, J., Han, X.H., Shi, Y.K., Lu, S.H., 2009. Isolation and identification of cancer stem-like cells in esophageal carcinoma cell lines. Stem Cells Dev., 18(3):465-474.

[14]Krejs, G.J., 2010. Gastric cancer: epidemiology and risk factors. Dig. Dis., 28(4-5):600-603.

[15]Leung, E.L., Fiscus, R.R., Tung, J.W., Tin, V.P., Cheng, L.C., Sihoe, A.D., Fink, L.M., Ma, Y., Wong, M.P., 2010. Non-small cell lung cancer cells expressing CD44 are enriched for stem cell-like properties. PLoS One, 5(11):e14062.

[16]Li, H., Hamou, M.F., de Tribolet, N., Jaufeerally, R., Hofmann, M., Diserens, A.C., van Meir, E.G., 1993. Variant CD44 adhesion molecules are expressed in human brain metastases but not in glioblastomas. Cancer Res., 53(22):5345-5349.

[17]McDonald, S., Greaves, L., Gutierrez-Gonzalez, L., Rodriquez-Jstto, M., Deheragada, M., Leedham, S.J., Tayor, R.W., Lee, C.Y., Preston, S.C., Loveu, M., et al., 2008. Mechanisms of field cancerization in the human stomach: the expansion and spread of mutated gastric stem cells. Gastroenterology, 134(2):500-510.

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

[19]Oates, J.E., Grey, B.R., Addla, S.K., Samuel, J.D., Hart, C.A., Ramani, V.A.C., Brown, M.D., Clarke, N.W., 2009. Hoechst 33342 side population identification is a conserved and unified mechanism in urological cancers. Stem Cell Devel., 18(10):1515-1522.

[20]Parkin, D.M., Bray, F., Ferlay, J., Pisani, P., 2005. Global cancer statistics. CA: Cancer J. Clin., 55(2):74-108.

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

[22]Ricci-Vitiani, L., Lombardi, D.G., Pilozzi, E., Biffoni, M., Todaro, M., Peschle, C., de Maria, R., 2007. Identification and expansion of human colon-cancer-initiating cells. Nature, 445(7123):111-115.

[23]Shi, Z., Bai, R., Fu, Z.X., Zhu, Y.L., Wang, R.F., Zheng, S., 2012. Induced pluripotent stem cell-related genes influence biological behavior and 5-fluorouracil sensitivity of colorectal cancer cells. J. Zhejiang Univ.-Sci. B (Biomed. & Biotechnol.), 13(1):11-19.

[24]Song, Z., Yue, W., Wei, B., Wang, N., Li, T., Guan, L., Shi, S., Zeng, Q., Pei, X., Chen, L., 2011. Sonic hedgehog pathway is essential for maintenance of cancer stem-like cells in human gastric cancer. PLoS One, 6(3):e17687.

[25]Takaishi, S., Okumura, T., Tu, S., Wang, S.S., Shibata, W., Vigneshwaran, R., Gordon, S.A., Shimada, Y., Wang, T.C., 2009. Identification of gastric cancer stem cells using the cell surface marker CD44. Stem Cells, 27(5):1006-1022.

[26]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.

[27]Zhang, C., Li, C., He, F., Cai, Y., Yang, H., 2011. Identification of CD44⁺CD24⁺ gastric cancer stem cells. J. Cancer Res. Clin. Oncol., 137(11):1679-1686.

[28]Zhou, S., Schuetz, J.D., Bunting, K.D., Colapietro, A.M., Sampath, J., Morris, J.J., Laqutina, I., Grosvld, G.C., Osawa, M., Nakauchi, H., et al., 2001. The ABC transporter Bcrp1/ABCG2 is expressed in a wide variety of stem cells and is a molecular determinant of the side-population phenotype. Nat. Med., 7(9):1028-1034.