天花粉蛋白对结肠癌细胞株CMT-93凋亡及增殖的影响  

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

作　　者：周琳[1] 刘菲[1] 周光炎[1] 路丽明[1] 

机构地区：[1]上海交通大学医学院上海市免疫学研究所,上海200025

出　　处：《上海交通大学学报（医学版）》2014年第3期257-262,共6页Journal of Shanghai Jiao tong University:Medical Science

基　　金：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)~~

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(pAkt473和pAkt 308蛋白表达)。结果与对照组比较,5.0μg/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。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 p.g/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.

关 键 词：天花粉蛋白 结肠癌 细胞凋亡 细胞增殖 

分 类 号：R735.35[医药卫生—肿瘤]