Study on Biological Characteristics of CD133 Positive Population in Gastric Cancer Cells by Magnetic Activated Cell Sorting In Vitro_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

LU Ruiqi ¹ , WU Jugang ¹ , ZHOU Guocai ^1,2 , JIANG Haiguang ^1,2 , NI Xiaochun ¹ , YU Jiwei ¹ ,  JIANG Bojian. ¹

1.Department of General Surgery，No.3 People’s Hospital， Shanghai JiaoTong University School of Medicine， Shanghai 201900， China;;
2.Graduate School, Bengbu Medical College, Bengbu 233030, Anhui Province, China;

Corresponding author：

JIANG Bojian., Email: jiang_md@hotmail.com

Keywords：

Tumor initiator cell; CD133; Gastric cancer

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ObjectiveTo detect the expressions of CD133 in four gastric cancer cell lines (KATO-Ⅲ, SGC7901, AGS, and MKN-45) and investigate the different biological characteristics of CD133 positive (CD133+) cells and CD133 negative (CD133-) cells in the KATO-Ⅲ cell lines. MethodsThe CD133 gene and protein expressions of four gastric cancer cell lines were evaluated by semiquantitative RT-PCR and Western blot, respectively. CD133+ cells in KATO-Ⅲ cell lines were isolated by magnetic activated cell sorting (MACS) and examined in morphology, growth characteristics, proliferation, and differentiation in vitro. The sensitivity of different concentrations (0.05, 0.10, 0.20, 0.50, and 1.00 mg/ml) 5-fluoropyrimidinedione (5-FU) were contrasted by drug sensitivity testing in vitro (Cell Counting Kit 8-assay) between CD133+ cells and CD133- cells. ResultsThe expressions of CD133 gene and protein in the KATO-Ⅲ cell lines were significantly higher than those in the other three cell lines (P lt;0.05). CD133+ cells produced spheroid colonies in serum-free medium culture and were ber abilities of proliferation and differentiation than those of CD133- cells in vitro. The inhibitor rate of the CD133+ cells at concentration of 0.50 mg/ml was lower than that of CD133- cells (P lt;0.05). ConclusionsCell population with CD133+ in the KATO-Ⅲ cell lines have b ability of cloning, better capability of proliferation and differentiation, as well as anticancer drug resistance to 5-FU. CD133 can be applied as one of surface markers for the detection to gastric cancer initiator cells.

Citation： LU Ruiqi,WU Jugang,ZHOU Guocai,JIANG Haiguang,NI Xiaochun,YU Jiwei,JIANG Bojian.. Study on Biological Characteristics of CD133 Positive Population in Gastric Cancer Cells by Magnetic Activated Cell Sorting In Vitro. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2011, 18(12): 1265-1270. doi: Copy

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15.	AlHajj M, Wicha MS, BenitoHernandez A, et al. Prospective identification of tumorigenic breast cancer cells [J]. Proc Natl Acad Sci USA, 2003, 100(7): 39833988.
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20.	张鹏, 俞继卫, 姜波健. 肿瘤干细胞与胃肠道癌发生和转移 [J]. 中国普外基础与临床杂志, 2009, 16(1): 8184.
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1. Bonnet D, Dick JE. Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hematopoietic cell [J]. Nat Med, 1997, 3(7): 730737.
2. Wulf GG, Wang RY, Kuehnle I, et al. A leukemic stem cell with intrinsic drug efflux capacity in acute myeloid leukemia [J]. Blood, 2001, 98(4): 11661173.
3. Pérez Castillo A, AguilarMorante D, MoralesGarcía JA, et al. Cancer stem cells and brain tumors [J]. Clin Transl Oncol, 2008, 10(5): 262267.
4. Field M, Alvarez A, Bushnev S, et al. Embryonic stem cell markers distinguishing cancer stem cells from normal human neuronal stem cell populations in malignant glioma patients [J]. Clin Neurosurg, 2010, 57: 151159.
5. Yu SC, Ping YF, Yi L, et al. Isolation and characterization of cancer stem cells from a human glioblastoma cell line U87 [J]. Cancer Lett, 2008, 265(1): 124134.
6. Miraglia S, Godfrey W, Yin AH, et al. A novel fivetransmembrane hematopoietic stem cell antigen: isolation, characterization,and molecular cloning [J]. Blood, 1997, 90(12): 50135021.
7. Kang MK, Kang SK. Tumorigenesis of chemotherapeutic drugresistant cancer stemlike cells in brain glioma [J]. Stem Cells Dev, 2007, 16(5): 837847.
8. O’Brien CA, Pollett A, Gallinger S, et al. A human colon cancer cell capable of initiating tumour growth in immunodeficient mice [J]. Nature, 2007, 445(7123): 106110.
9. Rappa G, Fodstad O, Lorico A. The stem cellassociated antigen CD133 (prominin1) is a molecular therapeutic target for metastatic melanoma [J]. Stem Cells, 2008, 26(12): 30083017.
10. 王晓辉, 李非.胰腺癌干细胞的研究进展 [J]. 中国普外基础与临床杂志, 2011, 18(9): 931935.
11. Smith LM, Nesterova A, Ryan MC, et al. CD133/prominin1 is a potential therapeutic target for antibodydrug conjugates in hepatocellular and gastric cancers [J]. Br J Cancer, 2008, 99(1): 100109.
12. Yu JW, Zhang P, Wu JG, et al. Expressions and clinical significances of CD133 protein and CD133 mRNA in primary lesion of gastric adenocacinoma [J]. J Exp Clin Cancer Res, 2010, 29(1): 141151.
13. 吴巨钢, 陆瑞祺, 张鹏, 等. 胃癌患者外周血CD133 mRNA检测及其临床意义 [J]. 中华胃肠外科杂志, 2010, 14(1): 6667.
14. Kelly SE, Di Benedetto A, Greco A, et al. Rapid selection and proliferation of CD133+ cells from cancer cell lines: chemotherapeutic implications [J]. PLoS One, 2010, 5(4): e10035.
15. AlHajj M, Wicha MS, BenitoHernandez A, et al. Prospective identification of tumorigenic breast cancer cells [J]. Proc Natl Acad Sci USA, 2003, 100(7): 39833988.
16. Kume H, Teramoto S, Isurugi K. Invivo dedifferentiation of keratinocytes to epidermal stem cells [J]. Lancet, 2002, 359(9305): 528529.
17. Takenobu H, Shimozato O, Nakamura T, et al. CD133 suppresses neuroblastoma cell differentiation via signal pathway modification [J]. Oncogene, 2011, 30(1): 97105.
18. Wang Z, Li Y, Ahmad A, et al. Pancreatic cancer: understanding and overcoming chemoresistance [J]. Nat Rev Gastroenterol Hepatol, 2011, 8(1): 2733.
19. Chen J, Emara N, Solomides C, et al. Resistance to platinumbased chemotherapy in lung cancer cell lines [J]. Cancer Chemother Pharmacol, 2010, 66(6): 11031111.
20. 张鹏, 俞继卫, 姜波健. 肿瘤干细胞与胃肠道癌发生和转移 [J]. 中国普外基础与临床杂志, 2009, 16(1): 8184.
21. Garvalov BK, Acker T. Cancer stem cells: a new framework for the design of tumor therapies [J]. J Mol Med, 2011, 89(2): 95107.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Study on Biological Characteristics of CD133 Positive Population in Gastric Cancer Cells by Magnetic Activated Cell Sorting In Vitro

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