张力蛋白1抑制非小细胞肺癌细胞增殖和侵袭

Tensin1 inhibits the proliferation and invasion of non-small cell lung cancer cells

目的:探讨张力蛋白1(tensin1,TNS1)在非小细胞肺癌(NSCLC)组织中的表达水平、与NSCLC患者预后的关系及其对NSCLC细胞生物学功能的影响。方法:收集56例NSCLC患者的临床病理资料,通过电话方式进行随访,了解患者生存情况。荧光实时定量PCR(qRT-PCR)检测56例NSCLC标本和对应癌旁组织中TNS1 mRNA表达水平,分析其与NSCLC患者临床病理特征的关系。采用Kaplan-Meier法绘制生存曲线,分析TNS1 mRNA与NSCLC患者预后的关系。采用qRT-PCR和蛋白质印迹法分别检测NSCLC细胞A549、H1299与正常肺支气管上皮16HBE细胞中TNS1 mRNA和蛋白表达水平;将A549、H1299细胞各分为2组,分别转染TNS1过表达质粒(pcDNA3.1/TNS1组)和相应的空载质粒(阴性对照pcDNA3.1组);MTT实验、克隆形成实验、细胞划痕、Transwell实验分别检测细胞增殖、迁移、侵袭能力;蛋白质印迹法检测细胞上皮间质转化相关分子蛋白水平。结果:与癌旁组织相比,NSCLC组织中TNS1 mRNA表达明显降低(P<0.05)。TNS1 mRNA表达量与NSCLC患者淋巴结转移及术后肿瘤转移相关(P<0.05),与年龄、性别、吸烟、病理分型、肿瘤大小、TNM分期无明显相关性(P>0.05)。TNS1 mRNA高表达患者的3年累积无病生存率和累积总体生存率均高于TNS1 mRNA低表达患者(P均<0.05)。A549和H1299细胞中TNS1 mRNA和蛋白表达量均明显低于16HBE细胞(P均<0.01)。与对照组相比,pcDNA3.1/TNS1组A549和H1299细胞增殖、迁移、侵袭能力降低,E-钙黏蛋白表达明显上调(P<0.01),波形蛋白表达明显下调(P<0.01)。结论:TNS1在NSCLC组织及细胞中表达下调,与NSCLC患者预后相关,可抑制NSCLC细胞增殖和迁移。

Objective:To investigate the expression of tensin1(TNS1)in non-small cell lung cancer(NSCLC)tissues,its correlation with prognosis of NSCLC patients and its effect on the biological function of NSCLC cells.Methods:The clinical data of 56 patients with NSCLC were collected.Follow-up using telephone interview was performed to confirm the survival time.Quantitative reverse transcription-PCR(qRT-PCR)was used to test the mRNA expression of TNS1 mRNA in 56 pairs of NSCLC tissues and adjacent normal tissues,and the relationships between expression of TNS1 mRNA and clinical features in NSCLC patients were analyzed.Survival curve was drawn using the Kaplan-Meier method,and the relationships between expression of TNS1 and prognosis in NSCLC patients were analyzed.The mRNA and protein expression levels of TNS1 in NSCLC cell A549,H1299 and normal lung bronchial epithelial 16HBE cells was detected by qRT-PCR and Western blotting.A549 and H1299 cells were divided into 2 groups,transfected with the expression plasmid of TNS1(pcDNA3.1/TNS1 group)and empty plasmid(negative control pcDNA3.1 group),respectively.The abilities of cell proliferation,migration and invasion were detected by MTT,clone formation assay,wound-healing assay,and Transwell migration test.The protein expression levels of epithelial-mesenchymal transition related markers were detected by Western blotting.Results:Compared with adjacent normal tissues,the mRNA expression of TNS1 in NSCLC tissues was significantly down-regulated(P<0.05).The expression of TNS1 mRNA was correlated with lymph node metastasis and postoperative tumor metastasis of NSCLC patients(P<0.05),while there were with no significant correlation among age,sex,smoking,pathological type,tumor size,and TNM stage(P>0.05).The 3-year tumor-free survival and overall survival rates of patients with high TNS1 mRNA expression were higher than those with low TNS1 expression(P<0.05).The mRNA and protein expression levels of TNS1 in A549 and H1299 cells were greatly lower than those in 16HBE cells(P<0.01).Compared with the control groups,pcDNA3.1/TNS1 groups in A549 and H1299 cells showed lower proliferation,migration and metastasis,increased expression of E-cadherin(P<0.01)and decreased expression of vimentin(P<0.01).Conclusion:NSCLC tissues and cells showed lower expression of TNS1,which was related to the prognosis of NSCLC patients.TNS1 could inhibit proliferation and metastasis of NSCLC cells.