抑制TLR4表达在雷公藤内酯醇增强胃癌BGC823细胞对5-FU敏感性中的作用  

Triptolide enhances the sensitivity of gastric cancer BGC823 cells to 5-FU by suppressing TLR4 expression

作　　者：耿静 张树贤[1] 孙运良 GENG Jing;ZHANG Shuxian;SUN Yunliang(Department of Gastroenterology,First Peoples Hospital of Lianyungang,Lianyungang,Jiangsu,222000,China)

机构地区：[1]连云港市第一人民医院消化内科,江苏连云港222000

出　　处：《中国中西医结合消化杂志》2022年第4期263-267,共5页Chinese Journal of Integrated Traditional and Western Medicine on Digestion

基　　金：连云港市卫生健康委指导性科研项目(No:ZD1903)。

摘　　要：目的:观察雷公藤内酯醇(TPL)是否可通过抑制Toll样受体4(TLR4)表达以增强胃癌BGC823细胞对5-FU的敏感性,并分析其相关的下游分子机制。方法:采用CCK-8法检测不同浓度TPL以及TPL联合5-FU对BGC823细胞的增殖的影响。将BGC823细胞分为对照组、5-FU组(仅予5-FU)、TPL+5-FU组以及脂多糖(LPS)+TPL+5-FU组;Hoechst33258染色观察细胞凋亡形态学变化,流式细胞仪检测细胞凋亡率,Western blot检测TLR4、磷酸化AKT(p-AKT)、Survivin、活性Caspase-3蛋白表达。结果:TPL可呈浓度-时间依赖性的抑制BGC823细胞增殖。与5-FU组比较,TPL联合5-FU后对BGC823细胞的抑制率明显增加(P<0.01),其5-FU的半数抑制浓度(IC_(50))显著降低(P<0.01)。与5-FU组比较,TPL+5-FU组凋亡细胞数和细胞凋亡率均显著增加(P<0.01);而LPS+TPL+5-FU组凋亡细胞数以及细胞凋亡率均较TPL+5-FU组显著下降(P<0.05)。TPL+5-FU组的TLR4、p-AKT、Survivin蛋白表达均较5-FU组下降,而活性Caspase-3蛋白表达增加;与TPL+5-FU组比较,LPS+TPL+5-FU组的p-AKT、Survivin蛋白表达增加,活性Caspase-3蛋白表达减少。结论:TPL具有增强BGC823细胞对5-FU敏感性的作用,其机制与抑制TLR4表达,进而阻断PI3K/AKT通路、下调Survivin表达并促进Caspase-3活化有关。Objective: To observe whether triptolide(TPL) could enhance the sensitivity of gastric cancer BGC823 cells to 5-FU by inhibiting the expression of Toll like receptor 4(TLR4), and analyze the related downstream molecular mechanisms. Methods: The CCK-8 method was used to detect the effects of different concentrations of TPL and TPL combined with 5-FU on the proliferation of BGC823 cells. BGC823 cells were divided into control group, 5-FU group, TPL+5-FU group and lipopolysaccharide(LPS) +TPL+5-FU group;Hoechst33258 staining was used to observe the apoptosis morphological changes and flow cytometry was used to detect the cell apoptosis rate. TLR4, phosphorylated Akt(p-Akt), Survivin and Caspase-3 protein expression were detected by Western blot. Results: TPL could inhibit the proliferation of BGC823 cells in a concentration-time dependent manner. Compared with the 5-FU group, the inhibition rate of the TPL combined with 5-FU group on BGC823 cells increased significantly(P<0.01), and the half inhibitory concentration(IC_(50)) of 5-FU decreased significantly(P<0.01). Compared with the 5-FU group, the number of apoptotic cells and apoptosis rate of the TPL+5-FU group were significantly increased(P<0.01);However, the number of apoptotic cells and cell apoptosis rate of the LPS+TPL+5-FU group were significantly lower than those of the TPL+5-FU group(P<0.05). The TLR4, p-Akt and Survivin protein expression of the TPL+5-FU group were higher than those of the 5-FU group, and the expression of active Caspase-3 protein increased. Compared with the TPL+5-FU group, the p-Akt and Survivin protein expression of the LPS+TPL+5-FU group increased, but active Caspase-3 protein expression reduced. Conclusion: TPL can enhance the sensitivity of BGC823 cells to 5-FU, its mechanism is related to inhibiting TLR4 expression, blocking the PI3 K/AKT pathway, down regulating Survivin expression and promoting Caspase-3 activation.

关 键 词：雷公藤内酯醇 Toll样受体4 胃癌 磷酸化AKT 活性Caspase-3 

分 类 号：R735.2[医药卫生—肿瘤]