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。
Objective：To investigate the exression of miR-34a on lung cancer and normal lung tissues,and the effect and mechanism of miR-34a in lung cancer cell invasion and migration.Methods： qPCR was used to detect the expression of miR-34a on lung cancer.miR-34a-mimic and miR-34a-inhibitor were used to overexpress and knockdown miR-34a.qPCR was used to detect the ef-fectiveness.Western blot was used to detect the expression of Snail after induced with miR-34a-mimic and miR-34a-inhibitor.Luciferase reporter gene was used to detect interaction between miR-34a and Snail.Transwell invasion assay was used to detect invasion ability after induced with miR-34a-mimic and miR-34a-inhibitor.Scratch assay was used to detect migration ability after induced with miR-34a-mimic and miR-34a-inhibitor.The expression of E-cadherin,Vimentin and Twist were detected by Western blot.Results： miR-34a expression was significantly reduced in lung cancer.With the stage of lung cancer progression,the expression of miR-34a reduced.With the differentiation of lung cancer progression,the expression of miR-34a decreased.Decreasing of miR-34a was associated with lung cancer lymph node metastasis.miR-34a-mimic and miR-34a-inhibitor could overexpress and knockdown miR-34a.miR-34a could regulate expression of Snail.Snail was the direct target of miR-34a;miR-34a could regulate the invasion ability of human lung carcinoma H1650 cells;miR-34a could regulate the migration of human lung carcinoma H1650 cells;miR-34a could regulate the expression of E-cadherin,Vimentin and Twist.Conclusion： miR-34a plays the role of tumor suppressor factor in lung cancer.miR-34a can regulate the invasion and migration ability of lung carcinoma H1650 cells by Snail induced EMT.
作者
刘行仁
白义凤
梁良
冯静
邓菲
LIU Xing-Ren BAI YI-Feng LIANG Liang FENG Jing DENG Fei(Department of Respiration, Sichuan Provincial People＇s Hospital, Sichuan Academy of Medical Sciences, Chengdu 610072, China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2017年第5期646-651,共6页
Chinese Journal of Immunology
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
