氯化甲基汞抗大鼠C6胶质瘤细胞的体外研究

Inhibitory effect of methylmercury chloride on rat C6 glioma cells in vitro

目的：通过体外实验探讨氯化甲基汞（MMC）抗大鼠C6胶质瘤细胞的作用。方法：体外培养C6胶质瘤细胞,分为空白对照组和MMC给药实验组（将0.08-10.00μmol.L^-1MMC按浓度梯度分为8组）。采用四甲基偶氮唑盐（MTT）比色法检测不同浓度MMC对体外培养C6胶质瘤细胞的增殖抑制和杀伤作用,采用流式细胞仪测定MMC对C6胶质瘤细胞凋亡和细胞周期的影响。结果：1.25、2.50、5.00和10.00μmol.L^-1的MMC在体外均可抑制C6胶质瘤细胞的增殖,C6胶质瘤细胞存活率明显低于对照组（P〈0.05）,增殖抑制作用随浓度的增加而增强。2.50、5.00和10.00μmol.L^-1MMC作用于C6胶质瘤细胞4、8、16和32 h后细胞存活率明显低于对照组（P〈0.05）,细胞杀伤作用随浓度的增加和时间的延长而增强。0.31、0.63、2.50、5.00和10.00μmol.L^-1MMC作用于C6胶质瘤细胞24h后细胞凋亡/坏死率明显高于对照组（P〈0.05）。1.25、2.50和5.00μmol.L^-1MMC作用于C6胶质瘤细胞72 h后,G0/G1期细胞百分率明显高于对照组（P〈0.05）,S期细胞百分率明显低于对照组（P〈0.05）。结论：MMC能够杀伤大鼠C6胶质瘤细胞,抑制增殖,诱导凋亡,具有应用于胶质瘤治疗的潜在价值。

Objective To study the inhibitory effect of methylmercury chloride （MMC） on rat C6 glioma cells in vitro. Methods The rat C6 glioma cells were cultivated in vitro and divided into control group and MMC-treated group （0. 08--10. 00 μmol · L^-1 MMC were divided into 8 groups with concentration gradient）. MTT assay was performed to evaluate the proliferation inhibitory effect and cytotoxicity effect of MMC with different concentrations on cultured rat C6 gliorna cells, and flow cytornetry was used to assess the effects of MMC treatment on cell apoptosis and cell cycle in rat C6 glioma cells. Results 1.25, 2.50, 5.00 and 10. 00 μmol · L^-1 MMC could inhibit the proliferation of cultured rat C6 glioma cells in vitro, the viabilities of MMC treated C6 glioma cells were significantly lower than those in control group （P〈0. 05）, the inhibitory effect was in a dose-dependent manner. The cell viabilities of C6 glioma cells treated with 2.50, 5.00 and 10. 00μmol · L^-1MMC for 4, 8, 16 and 32 h were significantly lower than those in control group （P〈0. 05）, the cytotoxicity effect was in a dose and time dependent manner. When C6 glioma cells were treated with 0.31, 0.63, 2.50, 5.00 and 10. 00μmol · L^-1 MMC for 24 h, the apoptosis/necrosis rates were significantly higher than those in control group （P〈0.05） ; when treated with 1.25, 2.50 and 5.00μmol · L^-1 MMC for 72 h, the rates of G0/G1 phase cell significantly increased compared with control group （P〈 0. 05） ; while cells into S phase were decreased （P〈 0.05）. Conclusion MMC has cytotoxicity effect on cultured rat C6 glioma cells in vitro, and can inhibit proliferation and induce apoptosis.