目的 探讨miR-320a对非小细胞肺癌(non-small cell lung cancer, NSCLC)细胞上皮细胞间质化过程的调控作用及其可能的作用机制。方法 选取2017-07至2018-12于菏泽市立医院住院并进行手术治疗的NSCLC患者60例,术中取切除的癌组织和癌旁组织,RT-PCR和Western Blot检测癌组织和癌旁组织miR-320a和FoxM1表达;miR-320a mimics、miR-320a NC和miR-320a inhibitor转染A549细胞,24 h后划痕实验和Transwell小室实验检测细胞迁移、侵袭能力,双荧光素酶报告基因系统预测miR-320a与FoxM1的靶向关系,Western Blot检测FoxM1、E-Cadherin和Vimentin蛋白表达。结果 癌旁组织内miR-320a的表达高于癌组织(P<0.001),FoxM1的阳性细胞率低于癌组织(P<0.001);miR-320a NC组细胞迁移和侵袭能力低于miR-320a mimics组(P<0.001),而高于miR-320a inhibitor组(P<0.001);miR-320a可靶向下调FoxM1的表达水平;FoxM1-WT A549细胞中,miR-320a mimics组细胞E-cadherin表达显著高于miR-320a NC组(P<0.001),Vimentin表达显著低于miR-320a NC组(P<0.001);miR-320a inhibitor组细胞E-cadherin表达显著低于miR-320a NC组(P<0.001),Vimentin表达显著高于miR-320a NC组(P<0.001)。结论 miR-320a在NSCLC中可能作为抑癌基因发挥作用,这种作用可能是通过靶向抑制FoxM1的表达,从而抑制NSCLC细胞的上皮细胞间质化过程而实现的。
Abstract
Objective To detect the regulatory effect of miR-320a on the epithelial-mesenchymal transition of non-small cell lung cancer cells and the possible mechanism.Methods Sixty patients with NSCLC who were hospitalized and received surgery in our hospital between July 2017 and December 2018 were selected as subjects. The resected cancer tissues and adjacent tissues were taken intraoperatively.The expressions of miR-320a and FoxM1in cancer tissues and adjacent tissues were detected by RT-PCR and Western blot. A549 cells were transfected with miR-320a mimics, miR-320a NC, and miR-320a inhibitor. Twenty-four hours after transfection, scratches and Transwellwere performed to test the migration and invasion ability of cells in each group. The dual luciferase reporter gene system was used to predict the targeting relationship between miR-320a and FoxM1. Western blot was used to detect the expressions of FoxM1, E-Cadherin and Vimentin proteins in each group of cells.Results The relative expression of miR-320a in adjacent tissues was significantly higher than that in cancerous tissues (P<0.001), and the positive cell rate of FoxM1 was significantly higher than in adjacent tissues (P<0.001). The expression of miR-320a in patients with TNM stage Ⅰ to Ⅱ was significantly higher than that in patients with Ⅲ to Ⅳ. The expression of miR-320a in poorly differentiated cancer tissues was significantly lower than that in moderately and highly differentiated cancer tissues. The migration and invasion abilityof cells in MiR-320a NC group was poorer than that of miR-320a mimics group (P<0.001), but better than that of miR-320a inhibitor group (P<0.001). The dual luciferase reporter gene proved that miR-320a could down-regulate FoxM1 expression. The expression of E-cadherin in miR-320a mimics FoxM1-WT A549 group was significantly higher than that in miR-320a NC group (P<0.001), but the expression of Vimentin was significantly lower (P<0.001). The expression of E-cadherin in inhibitor group was significantly lower than that in miR-320a NC group (P<0.001), but the expression of Vimentin was significantly higher (P<0.001).Conclusions miR-320a may play the role of a tumor suppressor gene in non-small cell lung cancer. This effect may be achieved via targeted inhibition of FoxM1 expression, thereby inhibiting the epithelial-mesenchymal transition process of non-small cell lung cancer cells.
关键词
非小细胞肺癌 /
miR-320a /
FoxM1 /
上皮细胞间质化
Key words
non-small cell lung cancer /
miR-320a /
FoxM1 /
epithelial-mesenchymal transition
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基金
山东省医药卫生科技发展计划项目(2018WS502)
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