组蛋白去乙酰化酶抑制剂SAHA体外抑制人卵巢癌SKOV3细胞的实验研究

SAHA inhibition of the human ovarian cancer cell line SKOV3 in vitro

目的:观察组蛋白去乙酰化酶抑制剂辛二酰苯胺异羟肟酸(suberoylanilide hydroxamic acid,SAHA)体外对人卵巢癌SKOV3细胞增殖和凋亡的作用,并探讨其可能机制。方法:体外应用SAHA作用于人卵巢癌SKOV3细胞,MTT法检测细胞增殖,流式细胞术检测细胞凋亡率。Western blot法检测细胞内组蛋白H4乙酰化水平,实时定量PCR方法检测p21、Bcl-2和Bax基因mRNA表达。结果:经SAHA作用后的人卵巢癌SKOV3细胞生长受到抑制,细胞凋亡增加(各实验组与对照组比较,均P<0.05),呈剂量依赖性;同时细胞内组蛋白H4乙酰化水平增加,p21基因和Bax基因mRNA表达增加(各实验组与对照组比较,均P<0.05),呈剂量依赖性,Bcl-2基因表达无明显变化(各实验组与对照组比较,均P>0.05)。结论:SAHA体外可抑制人卵巢癌SKOV3细胞增殖和诱导SKOV3细胞凋亡,提高组蛋白乙酰化水平,增加p21和Bax基因表达可能是其作用机制之一。

Objective： This study aimed to determine the effects of suberoylanilide hydroxamic acid （SAHA） on the cell proliferation and apoptosis of the human ovarian cancer cell line SKOV3. The possible underlying mechanisms were also investigated. Methods： The SKOV3 cells were treated with the histone deacetylase inhibitor SAHA. Cell growth was estimated by using the MTT assay, whereas the cell apoptosis rate was estimated by flow cytometry. Furthermore, Western blot analysis was used to detect the acetyl level ofhistone H4, whereas real-time PCR was used to detect the mRNA of p21, Bcl-2, and Bax. Results： After treatment with SAHA, cellu- lar growth was decreased, whereas cell apoptosis increased in SKOV3 cells in a concentration dependent manner （the different SAHA groups vs. the control group, P〈0.05）, Meanwhile, the acetyl level of histone H4 was increased. The mRNA of p21 and Bax were simi- larly increased in a concentration dependent manner （the different SAHA groups vs. the control group, P〈0.05）. By contrast, the mRNA of Bcl-2 was not significantly changed （the different SAHA groups vs. the control group, P〉0.05）. Conclusion： SAHA could effectively inhibit cellular proliferation and induce apoptosis in ovarian cancer SKOV3 cells in vitro. The increased acetyl level of the histones could enhance the p21 and Bax gene expression, as one of its mechanisms.