紫杉醇联合卡培他滨调控MAPK/mTOR信号抑制食管癌的机制研究

The mechanism of paclitaxel combined with capecitabine in inhibiting esophageal cancer by MAPK/mTOR signaling

目的探讨紫杉醇联合卡培他滨调控MAPK/mTOR信号抑制食管癌的机制研究。方法选取2017年1月—2018年1月本院收集的80例晚期食管癌患者给予紫杉醇联合卡培他滨治疗2疗程,评价化疗临床疗效。免疫组织化学法检测化疗前后患者食管癌组织内MAPK和mTOR表达。选取食管癌Ec-9706细胞给予紫杉醇联合卡培他滨培养24 h、48 h和72 h,细胞计数CCK-8法检测食管癌Ec-9706细胞的存活率;Capase-3活性试剂盒检测Capase-3活性;Western blot法检测患者食管癌组织内和食管癌Ec-9706细胞中磷酸化蛋白p-MAPK、p-AKT、mTOR、NF-kB、BCL-2和cleaved-Capase-3蛋白表达水平。结果食管癌患者应用紫杉醇联合卡培他滨给药化疗后疗效为65.00%(52/80)。免疫组化法结果显示食管癌患者化疗后MAPK含量显著上升,mTOR含量显著下降,差异有统计学意义(P<0.05)。Western blot结果显示,食管癌患者应用紫杉醇联合卡培他滨给药化疗后,患者组织内p-MAPK、NF-kB和cleaved-Capase-3蛋白含量显著增加,p-AKT、mTOR和BCL-2蛋白含量显著降低,差异有统计学意义(P<0.05)。给予紫杉醇联合卡培他滨治疗后,食管癌细胞株的细胞存活率随时间的延长明显降低,Caspase-3活性随时间的延长显著升高,食管癌EC9706细胞p-MAPK、NF-kB和cleaved-Capase-3蛋白含量随时间的延长呈显著增加,p-AKT、mTOR和BCL-2蛋白含量随时间的延长呈显著降低,差异有统计学意义(P<0.05)。结论紫杉醇联合卡培他滨可能通过上调MAPK/mTOR信号抑制食管癌的发生与发展。

Objective To investigate the mechanism of paclitaxel combined with capecitabine in inhibiting esophageal carcinoma by regulating MAPK/mTOR signaling.Methods Eighteen patients with advanced esophageal cancer collected from January 2017 to January 2018 were treated with paclitaxel and capecitabine for 2 courses to evaluate the clinical efficacy of chemotherapy.Immunohistochemistry was used to detect the expression of MAPK and mTOR in esophageal cancer tissues before and after chemotherapy.Select esophageal cancer Ec-9706 cells to give paclitaxel combined with capecitabine culture24 h,48 h and 72 h.Cytokine CCK-8 assay was used to detect the survival rate of esophageal cancer Ec-9706 cells;Capase-3 activity kit was used to detect Capase-3 activity.Western blot was used to detect the expression levels of phosphoproteins p-MAPK,p-AKT,mTOR,NF-kB,BCL-2 and cleaved-Capase-3 in esophageal cancer tissues and esophageal carcinoma Ec-9706 cells.Results The efficacy of chemotherapy with paclitaxel plus capecitabine was 65.00%(52/80)in esophageal cancer patients.The results of immunohistochemistry showed that the levels of MAPK in esophageal cancer patients increased significantly after chemotherapy,and mTOR levels decreased significantly(P<0.05).Western blot results showed that after chemotherapy of paclitaxel plus capecitabine in patients with esophageal cancer,p-MAPK,NF-kB and cleavedCapase-3 protein levels increased significantly in patients with p-AKT,mTOR and BCL-2.Protein content was significantly reduced,and the difference was statistically significant(P<0.05).After treatment with paclitaxel and capecitabine,the cell survival rate of esophageal cancer cell lines decreased significantly with time,the activity of caspase 3 increased significantly with time,and the protein levels of p-MAPK,NF-kB and cleaved-Capase-3 in esophageal cancer EC9706 cells increased significantly with time.the protein content of p-AKT,mTOR and BCL-2 was significantly decreased with time,and the difference was statistically significant(P<0.05).Conclusion Paclitaxel plus capecitabine inhibits the development of esophageal cancer by upregulating MAPK/mTOR signaling.