Kang Fei-wu, Wang Kai, Wu Min, Wang Zuo-lin, Zhu Yan, Min Rui
Dept. of Oral and Maxillofacial Surgery, Hospital of Stomatology, Tongi University, Shanghai 200072, China.
Hua Xi Kou Qiang Yi Xue Za Zhi. 2010 Oct;28(5):560-4.
To detect the expression of CD133 in human tongue squamous cell carcinoma Tea8113 cell line and observe proliferation ability of CD133 groups in vitro.
Limiting dilution assay was employed to observe the proliferating character of Tca8113 single cell in vitro. The ability of growing as cancer spheroids was observed with ultra-low attachment plates. The flow cytometry was used to detect the expression of putative tumor-initiating cell marker CD133 in human tongue squamous cell carcinoma Tca8113 cell line. The selective technique of immunomagnetic beads was applied to purify CD133 tumor cells, CD133 tumor cells were cultured and their ability of proliferation were observed in vitro.
After 12 days, the result of single cell culture in vitro revealed that about 5.23% of cultured Tca8113 cells possessed the capacity of continue proliferation. The cells line fromed floating clusters with one week of passaging cells into non-adherent plates. Approximately 0.95% of cells in Tca8113 cell line expressed CD133. Compared with CD133- cells and control Tca8113 cells, CD133+ cells demonstrated increased proliferation capacity. The proportion of CD133 cells decreased in culture as days passed. The percentage of CD133+ cells decreased from 92.45% to 1.62% in twelve days' culture.
Tumor stem cells have the character of heterogenity and lower proportion of CD133 but higher ability of proliferation, and the diferentiation in human tongue squamous cell carcinoma Tca8113 cell line in vitro, CD133 may be one of makers for tumor-initiating cell of human tongue squamous cell carcinoma Tca8113 cell line.
检测CD133在人舌鳞状细胞癌Tea8113细胞系中的表达,并观察CD133组体外增殖能力。
采用有限稀释法观察Tca8113单细胞体外增殖特性。用超低附着板观察形成癌球的能力。采用流式细胞术检测人舌鳞状细胞癌Tca8113细胞系中假定肿瘤起始细胞标志物CD133的表达。应用免疫磁珠分选技术纯化CD133肿瘤细胞,培养CD133肿瘤细胞并观察其体外增殖能力。
体外单细胞培养12天后,结果显示约5.23%的培养Tca8113细胞具有持续增殖能力。将细胞接种到无附着板中传代一周后形成漂浮细胞团。Tca8113细胞系中约0.95%的细胞表达CD133。与CD133-细胞和对照Tca8113细胞相比,CD133+细胞增殖能力增强。培养过程中CD133细胞比例随时间下降。培养12天后,CD133+细胞百分比从92.45%降至1.62%。
肿瘤干细胞具有异质性,CD133比例较低但增殖能力较强,且在人舌鳞状细胞癌Tca8113细胞系体外分化过程中,CD133可能是人舌鳞状细胞癌Tca8113细胞系肿瘤起始细胞的标志物之一。