MEX3A对肺鳞癌细胞增殖和转移的作用及其机制

Effect and mechanism of MEX3A on proliferation and metastasis of lung squamous cell carcinoma

目的研究肌肉过量RNA结合家族成员A(muscle excess RNA-binding family member A,MEX3A)表达在肺鳞癌组织的临床意义,并探讨MEX3A对肺鳞癌细胞增殖和转移能力的作用及分子机制。方法采用GEPIA在线软件分析MEX3A在肺鳞癌组织中的表达水平。收集住院的肺鳞癌和癌旁组织标本各58例,采用免疫组化检测MEX3A蛋白在肺鳞癌组织和癌旁组织中的表达水平,统计分析MEX3A表达与患者临床病理参数及预后的关系。构建MEX3A干扰及MEX3A过表达质粒,常规培养肺鳞癌细胞株H266,分为NC组(5μl Lip 2000和5μg对照质粒)、sh-MEX3A组(5μl Lip 2000和5μg MEX3A敲减质粒)和oe-MEX3A组(5μl Lip 2000和5μg MEX3A过表达质粒)。CCK-8法检测各组细胞增殖能力,Transwell实验检测各组细胞转移能力,Western blot检测各组细胞PI3K/AKT信号通路PI3K、p-PI3K、AKT和p-AKT蛋白的表达。结果GEPIA分析和免疫组化结果显示与癌旁组织相比,MEX3A在肺鳞癌组织中的表达显著上调(P<0.05)。MEX3A高表达与患者淋巴结转移和TNM分期相关(P<0.05);与MEX3A低表达肺鳞癌患者相比,MEX3A高表达肺鳞癌患者预后较差(P<0.05)。与NC组相比,sh-MEX3A组肺鳞癌细胞增殖和转移能力显著下降(P<0.05),oe-MEX3A组肺鳞癌细胞增殖和转移能力显著增加(P<0.05);与NC组相比,sh-MEX3A组肺鳞癌细胞PI3K/AKT信号通路关键蛋白p-PI3K和p-AKT的表达降低(P<0.05),oe-MEX3A组肺鳞癌细胞PI3K/AKT信号通路关键蛋白p-PI3K和p-AKT的表达增加(P<0.01)。结论MEX3A促进肺鳞癌恶性进展,可望作为肺鳞癌的新型生物标志物和治疗靶点。

Objective To study the clinical significance of muscle excess RNA-binding family member A(MEX3A)in lung squamous carcinoma tissues,and explore the effect of MEX3A on the proliferation and the metastasis of lung squamous carcinoma cell and its molecular mechanism.Methods GEPIA online software was used to analyze the expression of MEX3A in lung squamous carcinoma tissues.The expression level of MEX3A protein was detected in 58 cases of lung squamous carcinoma cell and 58 cases of adjacent tissues from hospitalized patients by immunohistochemistry,and the relationships between MEX3A expression and clinical pathological parameters,prognosis of patients were analyzed.MEX3A interfering and MEX3A overexpression plasmids were constructed.Lung squamous carcinoma H266 cells were routinely cultured,and divided into NC group(5μl Lip 2000 and 5μg control plasmid),sh-MEX3A group(5μl Lip 2000 and 5μg MEX3A knockdown plasmid)and oe-MEX3A group(5μl Lip 2000 and 5μg MEX3A overexpresion plasmid).CCK-8 was used to detect the cell proliferation ability,Transwell experiment was used to detect the invasion ability of cells,and Western blot was used to detect the protein expression of PI3K,p-PI3K,AKT and p-AKT in the PI3K/AKT signaling pathway.Results GEPIA analysis and immunohistochemistry results showed that the expression of MEX3A in lung squamous carcinoma tissues was significantly higher than that in the adjacent tissues(P<0.05).High expression of MEX3A was correlated with lymph node metastasis and TNM staging(P<0.05).Compared with low MEX3A expression of lung squamous carcinoma patients,the high MEX3A expression of lung squamous carcinoma patients had a worse prognosis(P<0.05).Compared with NC group,the proliferation and invasion abilities of lung squamous cell carcinoma cells were decreased significantly in sh-MEX3A group(P<0.05),and the proliferation and invasion abilities of lung squamous cell carcinoma cells were increased significantly in oe-MEX3A group(P<0.05).Compared with NC group,the expression levels of p-PI3K and p-AKT were decreased in sh-MEX3A group(P<0.05),and the expression levels of p-PI3K and p-AKT in lung squamous cell carcinoma cell were increased in oe-MEX3A group(P<0.05).Conclusion MEX3A can promote the malignant progression of lung squamous cell carcinoma,and it is expected to be a new biomarker and therapeutic target for lung squamous carcinoma.