TM4SF1与肺癌细胞JAK2-STAT3信号通路相互作用机制及其在NSCLC中的表达

Interaction mechanism between four transmembrane protein 1 and JAK2-STAT3 signaling pathway in lung cancer cells and analysis of its expression in non-small cell lung cancer

摘要:

目的：探讨四次跨膜蛋白1（four transmembrane protein 1，TM4SF1）在非小细胞肺癌（non-small cell lung cancer，NSCLC）中的表达情况及其与NSCLC患者临床病理特征之间的关系。构建高表达TM4SF1的慢病毒表达载体稳定转染肺癌A549细胞，研究TM4SF1与肺癌细胞JAK2-STAT3信号通路的相互作用机制。方法：收集61例NSCLC患者手术切除的癌组织、相应配对的癌旁组织及因良性肺部疾病切除的正常肺组织10例。采用qRT-PCR及Western blot检测NSCLC组织、癌旁组织及正常肺组织中TM4SF1的表达水平。利用慢病毒转染技术在肺癌A549细胞中高表达TM4SF1基因，应用qRT-PCR及Western blot分析TM4SF1基因及JAK2-STAT3信号通路下游Sox2基因的表达水平，采用Transwell检测TM4SF1高表达对A549迁移的影响。结果：癌组织中TM4SF1 mRNA及蛋白表达水平明显高于癌旁组织（P<0.05）及正常肺组织（P<0.05）；TM4SF1的表达与患者年龄及性别无明显关联，与肿块的大小、分化程度、淋巴结转移、临床分期相关（P<0.05）。成功在肺癌细胞株中高表达TM4SF1，高表达TM4SF1后对A549细胞的迁移能力明显增强，并且上调JAK2-STAT3信号通路下游Sox2表达水平。结论：TM4SF1在NSCLC中呈现高表达，过表达该基因可以促进A549细胞的迁移并且上调JAK2-STAT3信号通路下游Sox2表达水平。TM4SF1可能通过该通路促进NSCLC的发生发展及远处转移，TM4SF1可能成为NSCLC的潜在治疗靶点。

Abstract:

Objective：To investigate the expression of four transmembrane protein 1（TM4SF1） in non-small cell lung cancer（NSCLC） and its relationship with clinicopathological features in NSCLC patients，to construct TM4SF1 lentiviral expression vector for stably transfecting lung cancer A549 cells，and to investigate the mechanism of interaction between TM4SF1 and JAK2-STAT3 signaling pathway in lung cancer cells. Methods：Resected cancer tissue and corresponding adjacent paracarcinoma of 61 patients with NSCLC and resected benign lung tissue of 10 patients were collected. Real-time immunofluorescence quantitative PCR（qRT-PCR） and Western blot were used to detect the expression of TM4SF1 in NSCLC tissues，paracancerous tissues and normal lung tissues. Lentiviral transfection was used to detect highly expressed TM4SF1 gene in lung cancer A549 cells. The qRT-PCR and Western blot were used to analyze the expression level of TM4SF1 gene and Sox2 gene. Transwell was used to examine the effect of high-expression TM4SF1 on A549 migration. Results：The expression of TM4SF1 mRNA and its protein in cancer tissues was significantly higher than that in adjacent tissues（P<0.05） and in normal lung tissues（P<0.05）；TM4SF1 expression was not associated with age and gender，but was associated with tumor size，degree of differentiation，lymph node metastasis，and clinical stage（P<0.05）. High-expressing TM4SF1 in lung cancer A549 cell line was successfully constructed，which was able to significantly enhance the migration ability of A549 cells and up-regulate the expression level of Sox2 in the downstream of AK2-STAT3 signaling pathway. Conclusion：TM4SF1 is highly expressed in NSCLC and overexpression of this gene can promote the migration of A549 cells and up-regulate the expression level of Sox2 in the downstream of JAK2-STAT3 signaling pathway. TM4SF1 may promote the development and distant metastasis of NSCLC through this pathway，and TM4SF1 may become a potential therapeutic target for NSCLC.