雷帕霉素对人体舌癌细胞增殖、迁移及LRG-1信号通路的作用机制研究

Mechanism of rapamycin on the proliferation,migration and LRG-1 signaling pathway of human tongue cancer cells

目的:探讨雷帕霉素对人体舌癌细胞增殖、迁移及LRG-1信号通路的作用机制研究。方法:将人舌鳞状细胞癌Tca8113细胞分为舌癌组和干预组,舌癌组不做任何处理,干预组采用不同浓度雷帕霉素处理,显微镜下观察Tca8113细胞形态、MTT检测其增殖、Transwell小室检测迁移,免疫组化检测LRG-1阳性表达,免疫组化检测LRG-1蛋白水平。结果:舌癌组Tca8113细胞互相积堆,排列密切生长,数目较多,给予雷帕霉素药物干预后,随着浓度的增加Tca8113细胞逐渐出现凋亡,细胞质出现泡状结构,细胞数目减少,异常形态的出现异常。浓度25μmol/L组在48 h、60 h及72 h细胞增殖较多,中高浓度(50、100μmol/L)细胞增殖较少,舌癌组Tca8113细胞增殖高于其他3组(P<0.05);中高浓度(50、100μmol/L)与舌癌组及25μmol/L组相差较大(P<0.05)。舌癌组Tca8113细胞中LRG-1呈强阳性表达,表达率在86%,中高浓度(50、100μmol/L)与舌癌组及25μmol/L组相比LRG-1阳性表达率较低(P<0.05);中高浓度(50、100μmol/L)与舌癌组及25μmol/L组相比LRG-1蛋白水平表达较低(P<0.05)。结论:雷帕霉素能够抑制人体舌癌细胞增殖,迁移能力降低,并与药物浓度及时间存在关联,雷帕霉素发挥作用可能与阻碍LRG-1信号通路表达相关。

Objective:To investigate the mechanism of rapamycin on the proliferation,migration and LRG-1 signaling pathway of human tongue cancer cells.Methods:The human tongue squamous cell carcinoma Tca8113 cells were divided into tongue cancer group and intervention group.The tongue cancer group did not do any treatment,and the intervention group was treated with multiple concentrations of rapamycin.Tca8113 cell morphology was observed with microscope,roliferation was detected by MTT,migration was detected by Transwell chamber,the positive expression of LRG-1 was detected by immunohistochemistry and LRG-1 protein level was detected by Western Blot.Results:After the intervention of rapamycin drugs,as the concentration increased,Tca8113 cells gradually appeared apoptosis,the cytoplasm appeared bubble-like structure,and the number of cells decreased.The 25μmol/L group had a higher degree of cell proliferation at 48 h,60 h and 72 h,with medium and high concentrations(50,100μmol/L)has less cell proliferation,and the Tca8113 cell proliferation in the tongue cancer group was higher than the other three groups(P<0.05).The medium and high concentration(50,100μmol/L)was different from the tongue cancer group and the 25μmol/L group(P<0.05).Compared with tongue cancer group and 25μmol/L group,the rate of LRG-1 positive expression and the LRG-1 protein level was lower(P<0.05)in the medium and high concentration(50,100μmol/L).Conclusion:Rapamycin can inhibit the proliferation of human tongue cancer cells,reduce the migration ability,and is related to drug concentration and time.The role of rapamycin may be related to the inhibition of LRG-1 signaling pathway expression.