As_4S_4对宫颈癌HeLa细胞增殖抑制和诱导凋亡的初步研究

The inhibition and induction of apoptosis in HeLa cell by As_4S_4

背景与目的:四硫化四砷(As4S4)是雄黄的主要成分。近年,实验显示雄黄治疗急性早幼粒细胞白血病(APL)疗效明显,发现其具有抗肿瘤作用,但对实体瘤宫颈癌未见报道,本实验体外研究中药雄黄主要成分As4S4对子宫颈癌HeLa细胞的作用,旨在观察其对HeLa细胞的增殖抑制和诱导凋亡的作用。方法:以不同浓度(7.5、15、30、60mmol/L)的As4S4,分12、24、36、48、60h处理HeLa细胞,用四甲基偶氮唑蓝(MTT)法测定细胞增殖抑制率;流式细胞仪(FCM)检测细胞凋亡率;用DNA梯状电泳(DNAladder)检测凋亡的发生。结果:不同浓度(7.5、15、30、60mmol/L)的As4S4处理的HeLa细胞,细胞增殖受到抑制,作用呈明显的时效和量效关系。24h时IC50为30mmo/L,与对照组相比,差异有非常显著性(P<0.01)。As4S4诱导HeLa细胞的凋亡率分别为(8.1±1.1)%、(29.6±2.5%)、(46±3)%、(62±4)%,与对照组比较(2.8±1.8)%,差异有极显著性(P<0.01),并呈浓度依赖性;DNAladder呈梯状条带。结论:As4S4对HeLa细胞体外具有增殖抑制和促进凋亡作用。

Background and purpose： Tetra-arsenic tetra-sulfide （As4S4） is a major component in realgar. In recent years, a series of studies had been undertaken both in vitro and in vivo, and demonstrated that As4S4 is effective for the treatment of cancer such as Acute Promyelocytic Leukemia （ APL）, but the efficacy of the agent is not well studied for the treatment of cervical cancer. The study was to investigate the impact of As4S4 on HeLa cells in terms of cell growth inhibition and apoptosis. Methods： HeLa cells were incubated with As4S4 for the varieties of period （ 12, 24, 36, 48, 60 h） at different concentrations（7.5, 15, 30, 60 mmol/L）. Cell growth was measured by MTT. Apoptosis was detected by double staining fiowcytometry （FCM） and DNA Ladder. Results： After treated with different concentrations of As4S4 （7.5, 15, 30, 60mmol/L）, the cell growth was significantly suppressed in dose-and time-dependent manner, proliferative inhibition was statistically significant for the cells with and without the treatment of agent （P 〈0.01）. FCM analysis showed that As4S4 could markedly induce apoptosis, apoptotic rate for the cells exposure to different concentrations of As4S4 （7.5, 15, 30, 60 mmol/L） were （ 8.1 ± 1.1） %, （ 29.6 ± 2.5） %, （46 ± 3） %, （ 62± 4） %, respectively, there were significant differences compared to control group（ 2.8 ± 1.8） %, （ P 〈 0.01） ; DNA fragmentation gel analysis showed typical DNA ladder of apoptosis. Conclusions： As4S4 could inhibit the proliferation and induce apoptosis in human HeLa cells.