NSCLC患者恶性胸水中H19下调miR-203对肺癌细胞侵袭能力的影响  

H19 in malignant pleural effusion of NSCLC enhances the invasion ability of lung cancer cells by down-regulating miR-203

作　　者：刘帅妹 王妹 杜雨轩 程文亮 李洁 葛晓军 LIU Shuai-mei;WANG Mei;DU Yu-xuan;CHENG Wen-liang;LI Jie;GE Xiao-jun(Department of Clinical Laboratory,The Second Affiliated Hospital of Zunyi Medical University,Zunyi 563000,Guizhou,China)

机构地区：[1]遵义医科大学第二附属医院检验科,遵义563000

出　　处：《医学研究生学报》2022年第10期1069-1075,共7页Journal of Medical Postgraduates

基　　金：贵州省卫生健康委科学技术基金(gzwjkj2020-1-170)。

摘　　要：目的H19在非小细胞肺癌(NSCLC)的异常高表达是目前研究的热点。文中旨在探究NSCLC患者恶性胸水中H19的表达水平及其与miR-203的调控关系。方法收集2019年6至2021年5月遵义医科大学第二附属医院检验科132例NSCLC胸水标本。根据H19的相对表达水平,将患者分为H19高表达组(肿瘤/对照≥1.5)和H19低表达组(肿瘤/对照<1.5),健康供者为对照组。根据miR-203的相对表达水平,将患者分为miR-203高表达组(肿瘤/对照≥2.0)和miR-203低表达组(肿瘤/对照≤2.0)。在萤光素酶检测分析中,健康供者为空白对照组,未加入miR-203激活剂和miR-203抑制剂为阴性对照组,加入miR-203激活剂为miR-203激活剂组,加入miR-203抑制剂为miR-203抑制剂组。在EMT标记蛋白的变化分析,未加入miR-203抑制剂和H19抑制剂为阴性组,加入miR-203抑制剂和H19抑制剂为miR-203抑制剂+H19抑制剂组,加入miR-203抑制剂为miR-203抑制剂组。采用Western blot检测细胞内蛋白表达;CCK-8检测细胞增殖能力;Transwell检测细胞侵袭能力;流式细胞术检测细胞周期及凋亡等。结果H19高表达组的生存时间[(59.22±20.97)个月]较H19低表达组[(85.70±13.12)个月]明显缩短(P<0.05),即H19高表达与NSCLC的预后不良相关。而miR-203高表达组生存时间[(64.15±25.89)个月]较miR-203低表达组生存时间[(55.44±26.65)个月]明显延长(P<0.05)。相关性分析发现,H19与miR-203呈负相关(r=-0.66,P<0.01)。与阴性对照组比较,miR-203抑制剂组能抑制上皮标志物CDH1的表达,并促进间质标志物SNAI1和Vimentin的表达(P<0.05)。与阴性对照组比较,miR-203抑制剂组和H19抑制剂组能促进上皮标志物CDH1的表达,且抑制间质标志物SNAI1和Vimentin的表达(P<0.05)。结论H19的表达能负向调控miR-203的表达,抑制H19的表达降低NSCLC细胞的增殖、侵袭,H19是NSCLC的调控机制之一。Objective The abnormal overexpression of H19 in NSCLC is a hot research topic.The aim of this study was to investigate the expression level of H19 in malignant pleural fluid of NSCLC patients and its regulatory relationship with miR-203.Methods 132 samples of malignant pleural fluid of NSCLC patients admitted to the Department of Clinical Laboratory of the Second Affiliated Hospital of Zunyi Medical University from June 2019 to May 2021 were collected.According to the relative expression level of H19,patients were divided into the H19 high expression group(tumor/control≥1.5)and the H19 low expression group(tumor/control<1.5),and healthy donors were the control group.According to the relative expression level of miR-203,patients were divided into the miR-203 high expression group(tumor/control≥2.0)and the miR-203 low expression group(tumor/control≤2.0).In the analysis of luciferase detection,the healthy donors were the blank control group,the group without miR-203 activator and miR-203 inhibitor were the negative control group,the group with miR-203 activator was the miR-203 activator group,and the group with miR-203 inhibitor was the miR-203 inhibitor group.In the analysis of changes of EMT labeled proteins,the group without miR-203 inhibitor and H19 inhibitor was the negative group,the group with miR-203 inhibitor and H19 inhibitor was the miR-203 inhibitor+H19 inhibitor group,and the group with miR-203 inhibitor was the miR-203 inhibitor group.Western blot was used to detect the expression of intracellular proteins;CCK-8 was used to detect cell proliferation;Transwell was used to detect cell invasion;Flow cytometry was used to detect cell cycle and apoptosis.Results Survival curve analysis showed that the overall survival(85.70±13.12)of the H19 low expression group(59.22±20.97)was long,while the overall survival of the miR-203 high expression group(64.15±25.89)and the lower expression group(55.44±26.65)were long;the correlation analysis found that H19 was inversely correlated with miR-203(r=-0.66,P<0.

关 键 词：非小细胞肺癌 H19 miR-203 恶性胸水 

分 类 号：R734.2[医药卫生—肿瘤]