TRB3对人肺腺癌A549细胞生物学行为影响的初步研究  

Effects of TRB3 on Biological Behavior of Human Lung Adenocarcinoma Cells

作　　者：周辉[1] 陈建华[1] 罗永忠[1] 易青[1] 易辉煌[1] 李芳[1] 肖玲[2] 

机构地区：[1]中南大学湘雅医学院附属肿瘤医院内一科,湖南长沙410013 [2]中南大学湘雅医学院组织学与胚胎学教研室,湖南长沙410013

出　　处：《肿瘤药学》2013年第6期442-446,共5页Anti-Tumor Pharmacy

基　　金：湖南省科技厅科技计划项目资助(No.2012SK3249);湖南省医药卫生科研计划课题项目资助(No.B2012-098)

摘　　要：目的探讨TRB3基因对人肺腺癌A549细胞生物学行为的影响。方法免疫组化及半定量RT-PCR测定NSCLC患者肿瘤组织中TRB3的表达,分析其与临床病理特征的相关性,构建shRNA TRB3重组质粒,脂质体法转染A549细胞,Transwell小室侵袭实验、RT-PCR、Western blot等方法探讨TRB3基因沉默后对A549细胞生物学行为的影响。结果①在所有类型的NSCLC中,尤其是腺癌中,TRB3表达上调。TRB3的表达与肿瘤大小、淋巴结转移、远处转移和复发相关,并与患者状况显著相关(P<0.05)。与自身癌旁组织相比,60例肺癌组织中TRB3 mRNA水平明显增高(P<0.001)。此外,Kaplan–Meier生存曲线表明TRB3表达与总体生存期和无病生存率呈负相关。②与对照组比较,shTRB3转染组TRB3表达明显降低。TRB3沉默后抑制A549细胞的生长。同时,Transwell侵袭实验显示空载体转染组与对照组之间无显著性差异。与其他两组相比,shTRB3转染组侵袭率明显降低(P<0.001)。③与空白载体组相比,shTRB3 A549细胞中Notch1基因表达明显下调(P<0.001),两者呈正相关。结论沉默TRB3表达可抑制肺癌细胞生长,降低其侵袭能力,TRB3有可能成为治疗肺癌,预测其预后的一个新靶点。Objective To investigate the influence of TRB3 on the biological behavior of human lung adenocarcinoma A549 cells. Methods The expression of TRB3 in NSCLC tissue was detected by immunochemistry and semi-quantitative R.T-PCtk, and its cor- relation with the clinical pathological features was analyzed, shP, NA TRB3 recombinant plasmid was constructed, and transfected into A549 cells. The biological behavior ofA549 cells was detected by Transwell chamber invasion assay, RT-PCR, Western blot, and so on. Results The expression of TRB3 was up-regulated in all types of NSCLC, especially in adenocarcinoma, but it is normal in normal lung tissue. It was related to tumor size, lymph node metastasis, distant metastasis, recurrence and the status of patients with NSCLC （P〈0.05）. Compared with the adjacent normal tissues, the level of TRB3 mRNA was significantly increased in the tissues of 60 cases with lung cancer （P 〈 0.001）. In addition, Kaplan - Meier survival curves showed that the expression of TRB3 was negatively correlated with the overall survival and disease-free survival （Fig. 1B, 1C）. （2） Compared with control group, the expression of TRB3 was significantly de- creased in shTRB3 transfected group. The growth ofA549 cells was inhibited after TRB3 knockdowned. At the sanle time, there was no significant difference between the empty vector transfected group and the control group. Compared with the other two groups, the inva- sion of the shTRB3 transfected group was significantly decreased （P〈 0.001）. （3）Compared with the empty vector group, the expression of Notchl in the shTRB3 transfected group was significantly down-regulated （P〈 0.001）, and there was a positive correlation between them. Conclusion The decreased expression of TRB3 may inhibit the growth of A549 cells, and reduce its invasiveness. TRB3 would become a new target for treatulent and prediction on the lung cancer prognosis.

关 键 词：TRB3 非小细胞肺癌 基因敲除 细胞侵袭 

分 类 号：R734.2[医药卫生—肿瘤]