miR-571靶向SLC1A5调控食管癌细胞TE-13的凋亡研究

Study on miR-571 targeting SLC1A5 to regulate the apoptosis of esophageal cancer cells TE-13

[目的]探讨miR-571调控食管癌细胞凋亡的潜在分子机制。[方法]纳入2019年1月~2020年5月我院收治的食管癌患者60例,分析miR-571在其癌组织和癌旁组织中的表达情况。在食道癌细胞系Eca-109、NEC、TE-5、TE-10、TE-11、TE-13、KYSE-30、KYSE-150、KYSE-450、KYSE-510中过表达或敲低miR-571,分析其凋亡水平。通过miRDB在线分析和荧光素酶报告系统检测miR-571的底物。[结果]miR-571在食管癌组织中的表达水平低于癌旁组织(P<0.05)。在多种食管癌细胞中过表达miR-571后凋亡水平均有不同程度的上升(P<0.05);而敲低miR-571后凋亡水平均有不同程度的下降(P<0.05)。敲低SLC1A5时,食管癌细胞TE-13的凋亡水平上升(P<0.05)。过表达SLC1A5后,管癌细胞TE-13的凋亡水平下降(P<0.05)。在食管癌细胞TE-13中敲低miR-571后,SLC1A5的mRNA水平和蛋白水平上升(P<0.05);而过表达miR-571后,SLC1A5的mRNA水平和蛋白水平下降(P<0.05)。同时,荧光素酶报告实验发现miR-571靶向SLC1A5的3’端非编码区(P<0.05)。同时敲低miR-571和SLC1A5后,食管癌细胞TE-13的凋亡水平无明显变化(P<0.05)。同时过表达miR-571和SLC1A5后,食管癌细胞TE-13的凋亡水平无明显变化(P<0.05)。[结论]miR-571通过靶向SLC1A5 mRNA的3’端非编码区,降低SLC1A5的翻译水平,并促进了食管癌细胞TE-13的凋亡。

[Objective]Explore the potential molecular mechanism of miR-571 regulating the apoptosis of esophageal cancer cells.[Method]Sixty patients with esophageal cancer admitted to our hospital from January 2019 to May 2020 were enrolled,and the expression of miR-571 in their cancer tissues and adjacent tissues was analyzed.Overexpression or knockdown of miR-in esophageal cancer cell lines Eca-109,NEC,TE-5,TE-10,TE-11,TE-13,KYSE-30,KYSE-150,KYSE-450,KYSE-510571,analyze the level of apoptosis.The substrate of miR-571 is detected by miRDB online analysis and luciferase reporter system.[Result]The expression level of miR-571 in esophageal cancer tissues was lower than that of adjacent tissues(P<0.05).In a variety of esophageal cancer cells overexpressing miR-571,the level of apoptosis increased to varying degrees(P<0.05);and the level of apoptosis decreased to varying degrees after miR-571 was knocked down(P<0.05).It was found that when SLC1 A5 was knocked down,the apoptotic level of TE-13 in esophageal cancer cells increased(P<0.05).After overexpression of SLC1 A5,the apoptotic level of tube cancer cell TE-13 decreased(P<0.05).After knocking down miR-571 in esophageal cancer cells TE-13,the mRNA and protein levels of SLC1 A5 increased(P<0.05);while overexpression of miR-571,the mRNA and protein levels of SLC1 A5 decreased(P<0.05).At the same time,the luciferase report experiment found that miR-571 targets the 3’non-coding region of SLC1 A5(P<0.05).After knocking down miR-571 and SLC1 A5 at the same time,the apoptotic level of TE-13 in esophageal cancer cells did not change significantly(P<0.05).After overexpression of miR-571 and SLC1 A5 at the same time,there was no significant change in the apoptotic level of TE-13 in esophageal cancer cells(P<0.05).[Conclusion]miR-571 reduces the translation level of SLC1 A5 by targeting the 3’non-coding region of SLC1 A5 mRNA,and promotes the apoptosis of TE-13 esophageal cancer cells.