[1]郑庆委,高淑娴,吕杰,等.虫草素对人舌癌细胞凋亡、自噬的影响及其分子机制[J].南方医科大学学报,2018,(04):390.
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虫草素对人舌癌细胞凋亡、自噬的影响及其分子机制()
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《南方医科大学学报》[ISSN:/CN:]

卷:
期数:
2018年04期
页码:
390
栏目:
出版日期:
2018-04-30

文章信息/Info

Title:
Effect of cordycepin on apoptosis and autophagy of tongue cancer cells in vitro and the molecular mechanism
作者:
郑庆委高淑娴吕杰陈登宇陈杰李慧慧管俊昌
关键词:
虫草素自噬凋亡增殖
Keywords:
cordycepin autophagy apoptosis proliferation
摘要:
目的研究虫草素对人舌癌TCA-8113细胞的细胞周期、凋亡和自噬的影响并探讨虫草素抑制舌癌发生的作用机制。方 法用CCK-8法检测虫草素对TCA-8113细胞增殖的作用;用流式细胞术检测不同浓度药物对细胞周期、细胞凋亡的影响;用荧 光定量PCR和Western blot 的方法检测凋亡相关基因Caspase-3,Caspase-9,Bcl-2、Bax;免疫组化方法检测自噬相关蛋白LC- 3β,P62,p-mTOR,AMPK。结果CCK-8 细胞增殖检测结果表明,虫草素能明显抑制TCA-8113 细胞的增殖,24 h 时IC50为 3.548 mg/mL;48 h时IC50为1.185 mg/mL;流式细胞结果显示,诱导细胞周期阻滞与S期,虫草素能显著的诱导TCA-8113细胞 凋亡,并且呈浓度依赖性。荧光定量PCR和Westen blot结果表明,虫草素可诱导促凋亡基因Bax、Caspase-3、Caspase-9的表达, 抑制抗凋亡基因Bcl-2的表达(P<0.05);免疫组化的结果表明,虫草素可促进LC-3β和AMPK的表达,抑制P62和p-mTOR的表 达(P<0.05)。结论虫草素对TCA-8113细胞的抑制并促进细胞凋亡,同时通过AMPK/mTOR通路诱导细胞自噬。
Abstract:
Objective To study the effect of cordycepin on cell cycle, apoptosis and autophagy of human tongue cancer TCA- 8113 cells and explore the mechanism of cordycepin for inhibiting the occurrence of tongue cancer. Methods CCK-8 method was used to assess the inhibitory effect of cordycepin on TCA-8113 cell proliferation in vitro. The cell cycle and cell apoptosis of TCA-8113 cells treated with different concentrations of cordycepin were analyzed using flow cytometry. The expressions of apoptosis-related genes caspase-3, caspase-9, Bcl-2, and Bax were examined using quantitative real-time PCR and Western blotting, and immunohistochemistry was used to detect the expressions of autophagy-related proteins LC-3β, P62, p-mTOR, and AMPK. Results CCK-8 assay showed that cordycepin significantly inhibited the proliferation of TCA-8113 cells in a concentration-dependent manner with an IC50 of 3.548 mg/mL at 24 h and an IC50 of 1.185 mg/mL at 48 h. Flow cytometric analysis showed that cordycepin caused cell cycle arrest at S phase and dose-dependently increased the apoptotic rate of TCA- 8113 cells. Treatment of the cells with cordycepin enhanced the expressions of Bax, caspase-3 and caspase-9 at both the mRNA and protein levels and inhibited the expression of the antiapoptotic gene Bcl-2. Immunohistochemistry demonstrated that cordycepin promoted the expression of LC-3β and AMPK and inhibited the expression of P62 and p-mTOR. Conclusion Cordycepin inhibits the proliferation and induces apoptosis of HCT-116 cells through the mitochondrial pathway and induces autophagy via the AMPK/mTOR pathway.

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更新日期/Last Update: 1900-01-01