lncRNA MALAT1/miR-141-3p/ZEB1分子轴调控胃癌SGC7901细胞的侵袭、迁移及上皮间质转化

lncRNA MALAT1/miR-141-3p/ZEB1 axis modulates invasion, metastasis and EMT of gastric cancer SGC7901 cells

目的:探究lncRNA MALAT1/miR-141-3p/ZEB1分子轴对胃癌(GC)SGC7901细胞侵袭、迁移及上皮间质转化(EMT的调控作用。方法:收集2014年4月至2017年5月武汉商职医院普外科手术切除的GC组织(非坏死部分)和配对癌旁组织(距肿瘤组织>5 cm)标本38例,同时选取正常胃上皮细胞GES1及GC细胞系SGC7901、HGC27、BGC823、MKN45和MKN28。qPCR实验检测MALAT1、miR-141-3p在GC组织和细胞系中的表达水平,CCK-8和Transwell实验检测敲降MALAT1对SGC7901细胞增殖、迁移和侵袭的影响,WB实验检测ZEB1、E-cadherin、N-cadherin和Vimentin的表达情况。双荧光酶素报告基因验证MALAT1、miR-141-3p和ZEB1的靶向关系,CCK-8和Transwell实验检测MALAT1/miR-141-3p/ZEB1分子轴对SGC7901细胞生物学行为的影响。结果:MALAT1在GC组织和细胞系中高表达(P<0.05或P<0.01)。敲降MALAT1显著抑制了SGC7901细胞增殖、迁移、侵袭及EMT(P<0.05或P<0.01);MALAT1与miR-141-3p、miR-141-3p与ZEB1均具有直接靶向关系;进一步研究表明,同时过表达miR-141-3p和MALAT1或ZEB1能够逆转miR-141-3p对SGC7901细胞生物学行为的抑制作用。结论:MALAT1通过靶向下调miR-141-3p对ZEB1的抑制作用,进而促进SGC7901细胞侵袭、迁移及EMT。

Objective: To investigate the regulatory effect of lncRNA MALAT1/miR-141-3 p/ZEB1 axis on the invasion, metastasis and epithelial mesenchymal transition(EMT) of gastric cancer(GC) cells. Methods: Thirty-eight pairs of GC tissues(non-necrotic part) and corresponding adjacent tissues(>5 cm away from tumor tissue) removed by general surgery in Wuhan Commercial Hospital from April 2014 to May 2017 were collected. Meanwhile, normal gastric epithelial GES1 cells and GC cell lines(SGC7901, HGC27,BGC823, MKN45 and MKN28) were selected. The expression level of MALAT1 and miR-141-3 p in GC tissues and cell lines were detected by qPCR. The effect of MALAT1 knockdown on proliferation, migration and invasion of SGC7901 cells was determined by CCK-8 assay and Transwell assay. WB was performed for measuring the expression level of ZEB1, E-cadherin, N-cadherin and Vimentin. Dual luciferase reporter gene assay was used to validate the relationship among MALAT1, miR-141-3 p and ZEB1. CCK-8 assay and Transwell assay were used to detect the effect of MALAT1/miR-141-3 p/ZEB1 axis on biological behaviors of SGC7901 cells. Results: MALAT1 was over-expressed in GC tissues and cell lines(P<0.05 or P<0.01). Knockdown of MALAT1 significantly inhibited the proliferation, migration, invasion and EMT of SGC7901 cells(P<0.05 or P<0.01). The results of dual luciferase reporter gene assay showed that MALAT1 directly targeted miR-141-3 p, as well as for miR-141-3 p and ZEB1. Further experiment indicated that simultaneous over-expression of miR-141-3 p and MALAT1 or ZEB1 could restore the biological behaviors of SGC7901 cells, which were inhibited by miR-141-3 p. Conclusion: MALAT1 promotes the invasion, metastasis and EMT of GC SGC7901 cells by down-regulating the inhibitory effect of miR-141-3 p on ZEB1.