藻蓝蛋白协同全反式维甲酸影响人宫颈癌HeLa细胞的增殖和凋亡

Effects of C-phycocyanin combined with all-trans retinoic acid on cervical cancer cells proliferation and apoptosis

目的:探讨全反式维甲酸(all-transretinoic acid,ATRA)和钝顶螺旋藻藻蓝蛋白(C-phycocyanin,C-PC)联合用药对人宫颈癌HeLa细胞繁殖和凋亡的影响及其诱导细胞凋亡可能的分子机制。方法:实验分4组:对照组,C-PC组,ATRA组,C-PC+ATRA联合用药组。MTT法检测ATRA和C-PC单独及联合用药对HeLa细胞增殖的影响并计算它们的IC50,TUNEL法检测单独及联合用药后HeLa细胞凋亡情况,免疫组化法检测Bcl-2的表达,Westren blotting法检测Caspase-3的表达。结果:ATRA和C-PC均具有抑制HeLa细胞增殖的作用,IC50分别为(0.158±0.036)mmol/L和(192.75±5.79)μg/L。采用不同浓度的ATRA分别联合40或80μg/L C-PC处理HeLa细胞,ATRA的IC50分别为(0.095±0.007)mmol/L和(0.062±0.004)mmol/L,明显低于ATRA单独用药时的IC50值;当达到相同的抑制率时,联合C-PC用药可以显著降低ATRA的使用剂量。与对照组相比,两种药物单独用药增加了HeLa细胞凋亡水平(IODC-PC=63.12,IODATRA=59.98,P<0.05);当两种药物联合用药时,凋亡水平增加更加显著(IOD=89.52,P<0.01)。两药联合使用后HeLa细胞显著下调Bcl-2和上调Caspase-3的表达水平(均P<0.01)。结论:ATRA和C-PC联合用药可抑制HeLa细胞增殖和诱导其凋亡,其分子机制可能是通过抑制Bcl-2表达、促进Caspase-3表达来实现的。

Objective： To investigate the effects of all-transretinoic acid（ ATRA） and C-phycocyanin（ C-PC）,either each alone or two in combination on cervical cancer cell growth and apoptosis in vitro. Methods： Immortalized human cervical cancer HeLa cells were treated with ATRA and C-CP,either each at various concentrations or two in various dose combinations. After treatment for 48 h,cell viability was assessed by MTT assay,apoptosis by TUNEL assay,and changes in Caspase-3 and Bcl-2 protein contents by Western blotting and immunohistochemistry staining,respectively. Results： CPC and ATRA each alone significantly inhibited the growth of HeLa cells; and IC50 was 0. 158 ± 0. 036 mmol /L for C-CP and 192. 75 ± 5. 79 μg /L for ATRA（ P 〈0. 05）. When ATRA was combined with C-PC,the IC50 was significantly lower than that when ATRA was used alone. Compared with non-treatment control,C-PC and ATRA each alone induced significant HeLa cell apoptosis（ P 〈0. 05）,and in the apoptotic effect was more pronounced when C-PC and ATRA were used together（ P 〈0. 01）. ATRA and C-PC each alone significantly decreased Bcl-2 protein content（ P 〈0. 01） but significantly increased Caspase-3 protein content（ P〈 0. 01） in Hela cells,and both changes became more significant when ATRT and C-CP were used together（ P〈 0. 01）. Conclusion： C-PC combined with ATRA may induce cervical cancer cellapoptosis by down-regulation of Bcl-2 gene and up-regulation of Caspase-3 gene more effectively than each alone.