组蛋白去乙酰化酶6抑制剂ACY1215对顺铂诱导肝癌细胞SMMC-7721凋亡的影响

Histone deacetylase 6 inhibitor ACY1215 enhanced cisplatin-induced apoptosis of hepatocellular carcinoma cells SMMC-7721

目的:观察组蛋白去乙酰化酶6(HDAC6)抑制剂ACY1215对顺铂诱导肝癌细胞SMMC-7721凋亡的影响及其机制。方法:分别应用生理盐水、顺铂和ACY1215联合顺铂处理SMMC-7721细胞24h,光镜下观察各组细胞生长情况,RT-PCR检测各组细胞HDAC6基因的表达,Western blotting分析各组细胞微管蛋白(α-tubulin)、乙酰化微管蛋白(Acetyl-α-tubulin)、凋亡相关蛋白(Bax、Bcl-2)及增殖相关蛋白Cyclin D1的表达。结果:ACY1215联合顺铂对SMMC-7721细胞增殖的抑制明显高于单用顺铂。RT-PCR结果显示ACY1215联合顺铂组细胞HDAC6表达水平低于顺铂组(P<0.05)。Western blotting结果显示,ACY1215联合顺铂组细胞促凋亡蛋白Bax表达量较顺铂组升高(P<0.05),而抑凋亡蛋白Bcl-2和增殖相关蛋白Cyclin D1较顺铂组降低(P<0.05);ACY1215联合顺铂组细胞Acetyl-α-Tubulin表达水平较顺铂组升高(P<0.05)。结论:ACY1215具有增强顺铂诱导肝癌细胞凋亡的作用。

Objective:To investigate apoptosis sensitization and mechanism of histone deacetylase 6 inhibitor ACY1215 on hepatocellular carcinoma cells SMMC-7721 induced by cisplatin. Methods:SMMC-7721 cells were treated with normal saline, cisplatin(DDP), DDP +ACY1215 for 24 hour. The growth of cells in each group was observed under light microscope;Reverse transcription-polymerase chain reaction(RT-PCR) was used to measure the gene expression of HDAC6 in each group;Western blotting was performed to detect the protein expression of Acetyl-α-tubulin, α-tubulin, Bax, Bcl-|2, Cyclin D1. Results:The proliferation inhibition of SMMC-7721 cells in DDP+ ACY1215 group was higher than DDP group. RT-PCR shown that the gene expression of HDAC6 in DDP+ ACY1215 group was lower than DDP group(P<0.05). Western blotting shown that DDP+ ACY1215 group resulted in the increase in proapoptotic protein Bax(P<0.05), the decrease in antiapoptotic proteins Bcl-2 and proliferative protein Cyclin D1(P<0.05). In addition, ACY1215 could enhance the acetylation of α-tubulin in cisplatin-induced cells. Conclusion:ACY1215 enhanced cisplatin-induced apoptosis of hepatocellular carcinoma cells.