上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） • 上一篇    下一篇

天花粉蛋白对结肠癌细胞株CMT-93凋亡及增殖的影响

周 琳，刘 菲，周光炎，路丽明   

  1. 上海交通大学医学院  上海市免疫学研究所， 上海 200025
  • 出版日期:2014-03-28 发布日期:2014-04-02
  • 通讯作者: 路丽明, 电子信箱: lulunew2002@aliyun.com。
  • 作者简介:周 琳（1989—）， 男， 硕士生; 电子信箱: 867386705@qq.com。
  • 基金资助:

    国家自然科学基金(31370904，30972691) ；上海市卫生系统新优青培养项目（2011XYQ015）；上海市科委基础研究重点项目（11JC1410802）

Effects of trichosanthin on apoptosis and proliferation of colorectal cancer cell line CMT-93

ZHOU Lin, LIU Fei, ZHOU Guang-yan, LU Li-ming   

  1. Shanghai Institute of Immunology, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
  • Online:2014-03-28 Published:2014-04-02
  • Supported by:

    National Natural Science Foundation of China, 31370904, 30972691; New Talents Training Plan of Shanghai's Health System, 2011XYQ015; Major Fundamental Research Program of Science and Technology Commission of Shanghai Municipality，11JC1410802

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摘要：

目的 探讨低剂量天花粉蛋白（TCS）对结肠癌细胞株CMT-93凋亡及增殖的影响。方法 以不同质量浓度的TCS对体外培养的CMT-93进行干预,设立未添加TCS的CMT-93作为对照组，利用实时细胞动态监测(RTCA)系统检测细胞增殖活性，流式细胞仪检测细胞增殖和凋亡，Real-Time PCR检测细胞凋亡相关基因的表达，Western blotting检测丝氨酸/苏氨酸蛋白激酶Akt和γ-H2AX蛋白表达以及Akt磷酸化水平（pAkt 473和pAkt 308蛋白表达）。结果 与对照组比较，5.0 μg/mL TCS干预能使CMT-93增殖活性降低，细胞凋亡率升高；促凋亡相关蛋白Bid、Bax、Bad mRNA表达上调；pAkt 473和pAkt 308蛋白表达减少，γ-H2AX蛋白表达增加。结论 TCS对结肠癌细胞株CMT-93具有诱导凋亡和抑制增殖的作用；其机制可能与TCS通过抑制Akt的活性，进而激活促凋亡相关蛋白诱导细胞发生凋亡，以及TCS的DNA损伤作用有关。

关键词: 天花粉蛋白, 结肠癌, 细胞凋亡, 细胞增殖

Abstract:

Objective To investigate effects of trichosanthin (TCS) on apoptosis and proliferation of colorectal cancer (CRC) cell line CMT-93. Methods CMT-93, a mouse colorectal cell line cultured in vitro, was treated by TCS of different mass concentrations. The control group was CMT-93 without being treated by TCS. The activity of cell proliferation was measured by the Real-Time Cellular Analysis (RTCA) system. The proliferation and apoptosis was detected by the flow cytometry. Real-Time PCR was used to evaluate gene expressions related to the apoptosis. Expressions of Akt and γ-H2AX and the phosphorylation level of Akt (pAkt 473 and pAkt 308)  were assayed by the Western blotting. Results Compared to the control group, TCS of 5.0 μg/mL significantly inhibited the cell viability of CMT-93 cells and induced apoptosis of CMT-93 cells. The mRNA levels of Bid, Bax, and Bad were up-regulated in TCS-treated group. The expressions of pAkt 473 and pAkt 308 decreased significantly after treated by TCS, while the expression of γ-H2AX increased. Conclusion The results suggest that TCS can induce the apoptosis and inhibit the proliferation of CMT-93 cells. The mechanism may be relevant to inhibiting the expressions of active proapoptotic related proteins of Akt and DNA damage of the TCS.

Key words: trichosanthin, colorectal cancer, cell apoptosis, cell proliferation