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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 combination of all-transretinoic acid (ATRA) and C-phycocyanin (C-PC) on the proliferation and apoptosis of human cervical cancer HeLa cells and its induction Possible molecular mechanism of apoptosis. Methods: The experiment was divided into 4 groups: control group, C-PC group, ATRA group and C-PC + ATRA combination group. The effects of ATRA and C-PC alone and in combination on the proliferation of HeLa cells were detected by MTT assay and their IC50 values were calculated. The apoptosis of HeLa cells was detected by TUNEL assay and the expression of Bcl-2 was detected by immunohistochemistry. blotting method was used to detect the expression of Caspase-3. Results: Both ATRA and C-PC inhibited HeLa cell proliferation with IC50 of (0.158 ± 0.036) mmol / L and (192.75 ± 5.79) μg / L, respectively. The IC50 of ATRA was (0.095 ± 0.007) mmol / L and (0.062 ± 0.004) mmol / L, respectively, which was significantly lower than that of ATRA treated with ATRA alone or in combination with 40 or 80 μg / L C-PC IC50 value; when the same inhibition rate is reached, combined C-PC administration can significantly reduce the dose of ATRA. Compared with the control group, the two drugs alone increased the level of apoptosis of HeLa cells (IODC-PC = 63.12, IODATRA = 59.98, P <0.05); when the two drugs were used in combination, the apoptosis increased more significantly = 89.52, P <0.01). HeLa cells significantly decreased Bcl-2 and up-regulated the expression of Caspase-3 in HeLa cells (both P <0.01). Conclusion: The combination of ATRA and C-PC can inhibit HeLa cell proliferation and induce apoptosis, and its molecular mechanism may be through inhibiting Bcl-2 expression and promoting Caspase-3 expression.