那可丁联合奥沙利铂对人舌鳞状细胞癌 Tca8113 细胞增殖及凋亡的影响

Effect of noscapine combined with oxaliplatin on proliferation and apoptosis of human tongue squamous cell carcinoma Tca-8113 cells

摘要:

目的 探究那可丁联合奥沙利铂对人舌鳞状细胞癌Tca8113 细胞增殖及凋亡的影响。方法 将 体外培养的人舌鳞状细胞癌Tca8113 细胞随机分为空白对照组、奥沙利铂组、那可丁组和联合用药组。 CCK-8 法检测各组药物对Tca8113 细胞的存活率；Hoechst33342 染色荧光显微镜下观察凋亡形态变化；流 式细胞术检测细胞凋亡率；Western blotting 法检测各组细胞中Bax、Bcl-2、PARP、Cleaved Caspase-9 及 Cleaved Caspase-3 蛋白的表达水平。结果 奥沙利铂组、那可丁组和联合用药组Tca8113 细胞存活率较空白 对照组低（P <0.05），联合用药组较奥沙利铂组、那可丁组低（P <0.05）。Hoechst33342 染色法结果显示：奥 沙利铂组、那可丁组和联合用药组处理后均使人舌鳞状细胞癌Tca8113 细胞发生凋亡特征性改变。奥沙利铂组、 那可丁组和联合用药组Tca8113 细胞凋亡率较空白对照组高（P <0.05），联合用药组较奥沙利铂组、那可丁 组高（P <0.05）。奥沙利铂组、那可丁组和联合用药组Bax、PARP、Cleaved Caspase-9 及Cleaved Caspase-3 的蛋白表达较空白对照组高（P <0.05），而Bcl-2 的蛋白表达较空白对照组低（P <0.05），联合用药组Bax、 PARP、Cleaved Caspase-9 及Cleaved Caspase-3 的蛋白表达较奥沙利铂组、那可丁组高（P <0.05），Bcl-2 的 蛋白表达较奥沙利铂组、那可丁组低（P <0.05）。结论 那可丁联合奥沙利铂对人舌鳞状细胞癌Tca8113 细 胞的生长具有协同抑制作用，并促其凋亡，其作用机制可能与调节细胞凋亡相关蛋白表达有关。

关键词:Tca8113/ 口腔鳞状细胞癌;那可丁;细胞增殖;细胞凋亡

Abstract:

Objective To investigate the effect of noscapine combined with oxaliplatin on proliferation and apoptosis of human tongue squamous cell carcinoma Tca8113 cells. Methods Tongue squamous cell carcinoma Tca8113 cells cultured in vitro were randomly divided into control group, oxaliplatin group, noscapine group and combination group. CCK-8 assay was used to determine cell viability. The morphological changes of apoptosis were observed by Hoechst33342 fluorescence staining. Apoptosis rate was detected by flow cytometry. Western blotting was used to detect the expression levels of Bax, Bcl-2, PARP, Cleaved Caspase-9 and Cleaved Caspase-3 in each group. Results The results of CCK-8 assay showed that the survival rate of Tca8113 cells in tongue squamous cell carcinoma in the combined treatment group was significantly lower than that in the noscapine group and oxaliplatin group (P < 0.05), noscapine group, oxaliplatin group and combined treatment group were significantly lower than that in the control group. The results of Hoechst33342 staining showed that the morphology of nuclei in each treatment group showed characteristic changes of apoptosis compared with the control group. Flow cytometry analysis indicated that noscapine group, oxaliplatin group, combined group noticeably induced apoptosis (P < 0.05). Western blotting analysis showed that the expressions of Bax, PARP, Cleaved Caspase-9 and Cleaved Caspase-3 protein were up-regulated and the expression of Bcl-2 protein was significantly down-regulated in oxaliplatin group and noscapine group compared with the control group; the expressions of Bax, PARP, Cleaved Caspase-9, Cleaved Caspase-3 in combination group were significantly higher compared with oxaliplatin group and noscapine group (P < 0.05); Bcl-2 in combination group was lower than that in oxaliplatin group and noscapine group (P < 0.05). Conclusions Noscapine and oxaliplatin has synergistic effect on inducing apoptosis of tongue squamous cell carcinoma Tca-8113 cells, the mechanism of which may be related to the regulation of apoptosis-related protein expression.

Key words:Tca8113/oral squamous cell carcinoma; noscapine; proliferation; apoptosis