【摘要】 目的 探究miR-140-3p靶向白介素8（CXCL8）对食管鳞癌（ESCC）细胞增殖、侵袭和迁移能力的影响。方法 将ESCC细胞系EC109细胞分为：miR-NC组（转染miR-NC）、miR-140-3p组（转染miR-140-3p mimic）、miR-140-3p+CXCL8-NC组（细胞共转染miR-140-3p mimic和pcDNA-NC）、miR-140-3p+CXCL8组（细胞共转染miR-140-3p mimic和pcDNA-CXCL8）,未做任何处理的EC109细胞记为NC组。双荧光素酶报告基因实验验证miR-140-3p、CXCL8的关系；qRT-PCR检测人EC109细胞和正常人食管鳞状上皮细胞系Het-1A中miR-140-3p表达；Western blot检测Het-1A、EC109细胞CXCL8蛋白以及上皮间质转化（EMT）相关蛋白水平；CCK8法检测EC109细胞活力；流式细胞术检测EC109细胞凋亡率；Transwell检测EC109细胞侵袭以及迁移细胞数量。结果 在EC109细胞中CXCL8呈高表达，miR-140-3p呈低表达；与NC组、miR-NC组相比，miR-140-3p组EC109细胞OD₄₅₀值、迁移、侵袭细胞数量、神经型钙粘附蛋白（N-cadherin）、波形蛋白（Vimentin）、CXCL8蛋白水平显著下降（P<0.05）,EC109细胞凋亡率、miR-140-3p水平、上皮钙粘附素（E-cadherin）蛋白水平显著升高（P<0.05）,而上调CXCL8减弱了过表达miR-140-3p抑制EC109细胞增殖、迁移、侵袭、EMT及促进其凋亡的效果；miR-140-3p负向调控CXCL8表达。结论 miR-140-3p通过下调CXCL8抑制ESCC细胞增殖、迁移和侵袭。更多还原

【Abstract】 Objective To investigate the effects of miR-140-3p on the proliferation, invasion, and migration ability of esophageal squamous cell carcinoma（ESCC）cells by targeting interleukin 8（CXCL8）.Methods ESCC cell line EC109 cells were separated into: miR-NC group（transfected with miR-NC）,miR-140-3p group（transfected with miR-140-3p mimic）,miR-140-3p+CXCL8-NC group（co-transfected with miR-140-3p mimic and pcDNA-NC）,miR-140-3p+CXCL8 group（co-transfected with miR-140-3p mimic and pcDNA-CXCL8）,and EC109 cells without any treatment were recorded as NC group.Dual luciferase reporter gene experiment was applied to verify the relationship between miR-140-3p and CXCL8;qRT-PCR was applied to detect the expression of miR-140-3p in human EC109 cells and normal esophageal squamous cell line Het-1A;Western blot was applied to detect the levels of CXCL8 protein and epithelial mesenchymal transition（EMT）related proteins in Het-1A and EC109 cells; CCK8 method was applied to detect EC109 cell viability; flow cytometry was applied to detect the apoptosis rate of EC109 cells; Transwell was applied to detect the invasion and migration of EC109 cells.Results CXCL8 was highly expressed and miR-140-3p was low expressed in EC109 cells; compared with the NC group and miR-NC group, the OD₄₅₀ value, migration and invasion cell counts, N-cadherin, vimentin, and CXCL8 protein levels of EC109 cells in the miR-140-3p group were obviously reduced（P<0.05）,the apoptosis rate, miR-140-3p level, and E-cadherin protein level of EC109 cells were obviously increased（P<0.05）,up-regulation of CXCL8 weakened the effect of overexpression of miR-140-3p on inhibiting EC109 cell proliferation, migration, invasion, EMT,and promoting apoptosis; miR-140-3p negatively regulated CXCL8 expression.Conclusion MiR-140-3p inhibits proliferation, migration, and invasion of ESCC cells by down-regulating CXCL8.更多还原