摘要
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.
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
北大核心
2014年第5期516-520,共5页
Chinese Journal of Cancer Biotherapy
基金
国家自然科学基金资助项目(No.81471546
No.81001346)
山东省医药卫生科技发展计划项目(No.2011HZ023)
青岛市公共领域科技支撑计划项目(No.2012-1-3-5-(4)-nsh)~~
关键词
全反式维甲酸
藻蓝蛋白
宫颈癌
HELA细胞
凋亡
增殖抑制
all-transretinoic acid
C-phycocyanin
cervical cancer
HeLa cell
apoptosis
proliferation inhibition