基于PI3K/AKT信号通路探讨表儿茶素对口腔癌细胞的作用

Effect of Epicatechin on Oral Cancer Cells Based on PI3K/AKT Signaling Pathway

目的探讨表儿茶素通过PI3K/AKT信号通路抑制人口腔癌KB细胞的作用。方法培养人口腔癌KB细胞,分为空白对照组、表儿茶素处理组(20、40、80、160 nmol/L),CCK-8法检测细胞增殖抑制率,ELISA法检测细胞上清TNF-α和IL-1β水平,RT-PCR法检测Bax和Bcl-2基因表达水平;Western blot法检测PI3K和AKT蛋白的表达水平。结果表儿茶素处理组24、48、72 h增殖抑制率高于空白对照组,差异有统计学意义(P<0.05),且80 nmol/L表儿茶素组和160 nmol/L表儿茶素组24、48、72 h增殖抑制率均高于20 nmol/L表儿茶素组,差异有统计学意义(P<0.05);表儿茶素处理组TNF-α和IL-1β水平低于空白对照组,差异有统计学意义(P<0.05),且80 nmol/L表儿茶素组和160 nmol/L表儿茶素组TNF-α和IL-1β水平低于20 nmol/L表儿茶素组,差异有统计学意义(P<0.05);表儿茶素处理组Bcl-2 mRNA表达量低于空白对照组,而Bax mRNA表达量高于空白对照组,差异有统计学意义(P<0.05),且80 nmol/L表儿茶素组和160 nmol/L表儿茶素组Bcl-2 mRNA表达量低于20 nmol/L表儿茶素组,而Bax mRNA表达量高于20 nmol/L表儿茶素组,差异有统计学意义(P<0.05);表儿茶素处理组PI3K和AKT蛋白表达水平低于空白对照组,差异有统计学意义(P<0.05),且80 nmol/L表儿茶素组和160 nmol/L表儿茶素组PI3K和AKT蛋白表达水平低于20 nmol/L表儿茶素组,差异有统计学意义(P<0.05)。结论表儿茶素可抑制人口腔癌KB细胞的增殖以及促进其凋亡,可能与抑制PI3K/AKT信号通路有关。

Objective To investigate the effect of epicatechin in inhibiting human oral cancer KB cells through PI3K/AKT signaling pathway.Methods Human oral carcinoma KB cells were cultured and divided into blank control group and epicatechin treatment groups(20,40,80,160 nmol/L).CCK-8 assay was used to detect the inhibition rate of cell proliferation.ELISA was used to detect the levels of TNF-αand IL-1βin cell supernatant.RT-PCR was used to detect the expression levels of Bax and Bcl-2 genes.The expression levels of PI3K and AKT proteins were detected by Western blot.Results The proliferation inhibition rates at 24,48 and 72 h in the epicatechin treatment group were higher than those in the blank control group,and the difference was statistically significant(P<0.05),while the proliferation inhibition rates at 24,48 and 72 h in the 80 nmol/L epicatechin group and 160 nmol/L epicatechin group were higher than those in the 20 nmol/L epicatechin group,and the difference was statistically significant(P<0.05).The levels of TNF-αand IL-1βin the epicatechin treatment group were lower than those in the blank control group,and the difference was statistically significant(P<0.05),while the levels of TNF-αand IL-1βin the 80 nmol/L epicatechin group and 160 nmol/L epicatechin group were lower than those in the 20 nmol/L epicatechin group,the difference was statistically significant(P<0.05).The expression of Bcl-2 mRNA in epicatechin treatment group was lower than that in blank control group,while the expression of Bax mRNA was higher than that in blank control group,and the difference was statistically significant(P<0.05),while the expression of Bcl-2 mRNA in 80 nmol/L epicatechin group and 160 nmol/L epicatechin group was lower than that in 20 nmol/L epicatechin group,the expression of Bax mRNA was higher than that in 20 nmol/L epicatechin group(P<0.05).The expression levels of PI3K and AKT in the epicatechin treatment group were lower than those in the blank control group,and the difference was statistically significant(P<0.05),while the expression levels of PI3K and AKT in the 80 nmol/L epicatechin group and 160 nmol/L epicatechin group were lower than those in the 20 nmol/L epicatechin group,and the difference was statistically significant(P<0.05).Conclusion Epicatechin can inhibit the proliferation of human oral cancer KB cells and promote their apoptosis,which may be related to the inhibition of PI3K/AKT signaling pathway.