TMPRSS4及Smac在胃癌侵袭和转移中的机制及临床意义的研究

目的:通过检测胃癌组织中TMPRSS4与Smac的表达情况,分析其在胃癌侵袭与转移中的机制,并探讨其在胃癌发生、发展过程中的作用与意义。方法:采用Western blot、RT-PCR及免疫组化的方法检测胃癌组织与正常胃组织的TMPRSS4与Smac的表达。用脂质体转染过表达TMPRSS4胃癌细胞,构建特异性TMPRSS4siRNA表达载体,通过Transwell侵袭实验与划痕愈合实验研究TM-PRSS4表达的胃癌细胞侵袭转移机制;采用脂质体介导转染胃癌细胞MKN-45,用丝裂霉素处理转染前后的胃癌细胞株MKN-45,观察MKN-45的凋亡情况。结果:TMPRSS4mRNA与蛋白在胃癌组织中的表达明显高于正常组织,且以中、高表达占多数,差异有统计学意义(P<0.01);TMPRSS4mRNA与蛋白的表达水平与TNM分期、淋巴结转移个数、肿瘤直径大小、浸润深度有关,差异均有统计学意义(P<0.05～0.01)。在正常胃组织中的Smac表达率明显高于胃癌组织,差异有统计学意义(P<0.01);胃癌细胞的浸润深度、淋巴结转移个数与Samc的表达相关,差异有统计学意义(P<0.05);肿瘤直径大小与TNM分期与Samc表达无明显相关,差异无统计学意义(P>0.05)。转染后的MKN-45对化疗药物的敏感性比转染前明显增强,差异有统计学意义(P<0.05)。随访后发现TMPRSS4与Samc和患者的预后及生存时间相关。结论:TMPRSS4与患者的预后相关,可为患者的预后判断提供一定的依据;Samc的过表达能提高化疗药物的敏感性,为判断临床药物治疗的疗效提供一定的依据。

Role of TMPRSS4 and Smac in gastric cancer invasion and metastasis and tis clinical significance

Objective: To detect the expression of TMPRSS4 in gastric cancer tissue and to explore its role in gastric cancer invasion and metastasis and tis clinical significance.Methods:Western,RT-PCR and blot immunohistochemical method were applied for detection of expression of TMPRSS4 and Smac,over-expressed TMPRSS4 gastric cancer cells were transfected by liposome method,TMPRSS4 siRNA expression vector were constructed,Transwell invasion assay and scratches healing experiment were used to explore the mechanism of gastric cancer cells invasion and metastasis.Liposomes mediated transfection of gastric cancer cell MKN-45 were conducted for mitomycin treatment and then observed the apoptosis.Results: Expression of TMPRSS4mRNA and protein in gastric carcinoma was significantly higher than that in normal tissue,majority of which were high or mediate expression showing significant difference(P<0.01).The expression levels of TMPRSS4mRNA and protein were relevant to TNM stage,number of lymph node metastasis,depth of invasion,diameter of tumor size(P<0.01).Expression rate of Smac in normal gastric tissue was significant higher than that in gastric carcinoma tissue(P<0.01);Smac expression was related with the number of lymph node metastasis,depth of invasion,but not related with diameter of tumor size or TNM stage(P>0.05).After transfection,MKN-45 cells were more sensitive to chemotherapeutic drugs than that before transfection.Follow-up show that TMPRSS4 and Samc were correlated with the prognosis and survival time of patients.Conclusion: TMPRSS4 is related with prognosis of gastric cancer patients indicating it provide hints for prognosis.Besides,over-expressed Samc can improve the sensitivity of chemotherapy drugs,which can help to evaluate clinical efficacy of drug therapies.