miR-34a通过Snail诱导肺癌EMT及促进其转移的分子机制

发布时间：2018-07-07 08:26

本文选题：miR-a + 肺癌　；参考：《中国免疫学杂志》2017年05期

【摘要】：目的:探讨miR-34a在肺癌组织中的表达情况以及miR-34a在肺癌细胞侵袭和迁移过程中的作用及其机制。方法:q PCR检测肺癌和正常肺组织中miR-34a的表达情况;使用miR-34a-mimic和miR-34a-inhibitor过表达和沉默miR-34a,q PCR检测沉默和过表达效果;Western blot检测沉默和过表达miR-34a后Snail蛋白的表达情况;荧光素酶报告基因检测miR-34a与Snail的相互作用;Transwell侵袭实验检测miR-34a的表达对肺癌细胞侵袭能力的影响;划痕实验检测miR-34a的表达对肺癌细胞迁移能力的影响;Western blot检测E-Cadherin、Vimentin和Twist蛋白的表达情况。结果:与正常肺组织相比,肺癌组织中miR-34a表达明显降低;且晚期、低分化和有淋巴结转移的肺癌组织miR-34a表达明显较早期、高分化和无淋巴结转移的肺癌组织低;miR-34a-mimic和miR-34a-inhibitor可以有效抑制和过表达miR-34a的表达;miR-34a能与Snail的3'UTR特异性结合;miR-34a可以调控肺癌H1650细胞的侵袭迁移能力;过表达miR-34a上调E-Cadherin,同时下调Vimentin和Twist蛋白的表达,沉默miR-34a则相反。结论:miR-34a在肺癌中表达明显降低,且跟肺癌分期分级以及淋巴结转移与否密切相关,同时miR-34a可以通过上皮间质转化调节肺癌细胞侵袭和迁移能力。
[Abstract]:Aim: to investigate the expression of miR-34a in lung cancer and the role and mechanism of miR-34a in invasion and migration of lung cancer cells. Methods the expression of miR-34a in lung cancer and normal lung tissues was detected by 1: Q PCR, and the overexpression of miR-34a-mimic and miR-34a-inhibitor was detected by using miR-34a-mimic and silencing miR-34aq PCR. The expression of snail protein was detected by Western blot after silencing and over-expression of miR-34a. The interaction between miR-34a and Snail was detected by luciferase reporter gene. The expression of miR-34a in lung cancer cells was detected by Transwell invasion assay. The effect of miR-34a expression on the migration ability of lung cancer cells was detected by scratch assay. The expression of Vimentin and Twist proteins in E-Cadherin blot was detected by Western blot. Results: the expression of miR-34a in lung cancer tissues was significantly lower than that in normal lung tissues, and the expression of miR-34a in lung cancer tissues with low differentiation and lymph node metastasis was significantly earlier in the late stage. Low miR-34a-mimic and miR-34a-inhibitor can effectively inhibit and overexpression the expression of miR-34a in lung cancer tissues with high differentiation and no lymph node metastasis. MiR-34a can specifically bind with Snail's 3UTR to regulate the invasion and migration ability of lung cancer H1650 cells. Overexpression of miR-34a up-regulated E-Cadherin and down-regulated the expression of Vimentin and Twist proteins, whereas silencing miR-34a down-regulated the expression of Vimentin and Twist proteins. Conclusion the expression of miR-34a is significantly decreased in lung cancer, which is closely related to the staging and lymph node metastasis of lung cancer, and miR-34a can regulate the invasion and migration of lung cancer cells through epithelial interstitial transformation.
【作者单位】：四川省医学科学院·四川省人民医院呼吸科;四川省医学科学院·四川省人民医院肿瘤科;四川省医学科学院·四川省人民医院中医科;四川省医学科学院·四川省人民医院肾内科;
【基金】：国家自然科学基金(No.81301910) 四川省卫计委科研课题(No.110137)资助
【分类号】：R734.2

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