硫化砷对人宫颈癌HeLa细胞增殖和凋亡及COX-2表达的作用

Effect of As4S4 on preliferation, apoptosis and COX-2 expression of human cervical carcinoma HeLa cells

目的:体外研究中药雄黄的主要成分硫化砷(As4S4)对子宫颈癌HeLa细胞的增殖抑制和诱导凋亡的作用及与环氧合酶-2(cyclooxygenase-2,COX-2)表达的关系。方法:以不同浓度的As4S4(7.5～60mg/L),分12～60h5个时间点处理HeLa细胞,四甲基偶氮唑蓝(MTT)法测定细胞增殖抑制率;DNA梯状电泳检测凋亡的发生;Western blot检测COX-2蛋白的表达;放射免疫法检测细胞前列腺素E2(prostaglandinE2,PGE2)释放水平。结果:As4S4可抑制HeLa细胞增殖,作用呈明显的时效和量效关系,P<0.01;As4S4可诱导HeLa细胞凋亡,DNA电泳呈梯状条带;As4S4可明显抑制COX-2的表达,P<0.01,并呈浓度依赖性;As4S4明显抑制HeLa细胞PGE2释放水平,其值分别为(70.56±2.03)、(48.58±2.28)、(29.25±1.57)和(18.02±1.04)μg/L,与对照组(83.15±1.01)μg/L比较差异有统计学意义,P<0.01。结论:As4S4体外对HeLa细胞具有增殖抑制和促进凋亡作用,其机制可能与抑制COX-2蛋白表达有关。

OBJECTIVE： growth inhibition and apoptosis To investigate the HeLa cell induced by As4S4 and the relationship with COX-2 expression in vitro. METHODS： HeLa cells were treated with various concentrations （7.5- 60 mg/L） of As4S4 for 12-60 h. The cell growth rate was measured by MTT; apoptosis was detected by DNA electrophoresis. The levels of PGE2 were measured by radioimmunoassay. The expression of COX-2 protein was also examined by Western blot analysis. RESULTS： After treated with different concentrations of As4S4 , the cell growth of HeLa was suppressed significantly in dose and time-dependent manner in proliferative inhibition rate, P〈0.01. As4S4 induced the apoptosis and DNA fragmentation gel analysis showed DNA ladder. As4S4 reduced the release of PGE2 in HeLa cells with the values being （70.56±2.03）,（48.58±2.28）, （29.25±1.57） and （18.02±1.04） μg/L, respectively, as compared with control group （83.15±0.01） μg/L, P〈0. 01. As4S4 inhibited the activity and expression of COX-2 with the rise of concentration, and it down-regulated the expression of COX-2 protein significantly, CONCLUSIONS： As4S4 can inhibit the proliferation and increase apoptosis in human HeLa cells, These effects may depend on the inhibition of the expression of COX-2 and level of PGE2 by As4 S4.