辛二酰苯胺异羟肟酸对人卵巢癌细胞恶性表型的抑制作用

The inhibitory effects of suberoylanilide hydroxamic acid on the malignant phenotypes of ovarian carcinoma cells

目的 探讨辛二酰苯胺异羟肟酸（SAHA）对人卵巢癌细胞恶性表型的作用及其可能的分子作用机制。方法（1）选取2组人卵巢癌细胞（SKOV3和SKOV3/DDP;HO8910和HO8910-PM）后,设置对照组和终浓度为1、3、5及7μmol/L SAHA组（SAHA 1~4组）,采用CCK-8法分别检测SAHA对细胞增殖的影响。（2）设置对照组和SAHA 1、2组（2和5μmol/L）,Annexin V-FITC/PI双标记流式细胞术分别检测SAHA对细胞凋亡及周期的影响。（3）RT-PCR和Western blot检测SAHA 1~4组表型相关蛋白的表达水平。结果 （1）随着SAHA浓度的增加,SKOV3细胞株SKOV3/DDP及HO8910细胞株48 h光密度（OD）值均呈逐渐降低趋势（均P〈0.05）。（2）48 h SAHA 1、SAHA 2组凋亡率高于对照组（均P〈0.05）;SAHA作用48 h,细胞周期发生阻滞,与对照组比较,SAHA 1组和SAHA 2组SKOV3和SKOV3/DDP细胞S期和G2/M期比例增高,HO8910和HO8910-PM细胞G0/G1期比例增高（P〈0.05）。（3）SAHA 1、2组Cyclin B1和Cdc2（p34）的m RNA表达水平均低于对照组,Caspase-3、p21和p53的m RNA表达水平明显高于对照组（P〈0.05）;SAHA 1~4组Ac-Histone H3和Ac-Histone H4和p53蛋白的表达较对照组明显增强,Cyclin B1、Cdc2（p34）蛋白的表达减弱。结论 SAHA通过调节恶性表型相关蛋白Caspase-3、p53、Cyclin B1和Cdc2（p34）的表达水平,促进组蛋白乙酰化,抑制卵巢癌细胞增殖,阻滞细胞周期并诱导细胞凋亡。

Objective To explore the effects and molecular mechanisms of suberoylanilide hydroxamic acid （SAHA） on ovarian carcinoma. Methods （I） Two groups of ovarian carcinoma cell lines （SKOV3 and SKOV3/DDP, HO8910 and HO8910-PM） were exposed to SAHA （1, 3, 5 and 7 μmol/L SAHA, group 1-group 4）. CCK-8 method was employed to evaluate the inhibitory effects of SAHA. （2）Ovarian cancer cell lines treated with SAHA （2 or 5 μmol/L SAHA） were used as 1 and 2 groups. Flow cytometry was performed following staining with Annexin V-FITC and PI for cell cycle and apoptosis. （3） Reverse transcription polymerase chain reaction （RT-PCR） and Western blot assay were used to assess the mRNA and protein expression levels of phenotypic correlation factor. Results （ 1 ）After 48 h of SAHA treatment, the OD value of SKOV3, SKOV3/DDP, and HO8910 showed a trend of gradually reduce （P 〈 0.05）. （2）The apoptotic rates were significantly higher in SAHA 1 and SAHA 2 groups than those of control group （P 〈 0.05）. Compared with control group, after 48 h of SAHA treat- ment, S phase and G2/M phase of SKOV3 and SKOV3/DDP cells increased; G0/G1 phase of HO8910 and HO8910-PM cells increased in SAHA 1 and 2 groups （P 〈 0.05）.（3）The expression levels of CyclinB1 and Cdc2 （p34） mRNA were significantly lower in SAHA 1 and 2 groups than those of control group, while the expression levels of Caspase-3, p21 and p53 mRNA expression were significantly higher in SAHA 1 and 2 groups than those of control group. Furthermore, the expression of Ac-Histone H3, Ac-Histone H4, p53 protein were markedly improved, and CyclinB 1, Cdc2（p34） protein decreased in SAHA 1- 4 groups. Conclusion SAHA may suppress cell growth, induce apoptosis and cause cycle arrest in ovarian carcinoma cells by promoting histone aeetylation or modulating their phenotype-related proteins of Caspase-3, p53, CyclinB 1 and Cdc2（p34）.