LncRNA BLACAT1调节miR-17-5p/DDX5轴对子宫内膜癌细胞恶性进展的影响

LncRNA BLACAT1 regulates the effect of miR-17-5p/DDX5 axis on the malignant progression of endometrial cancer cells

目的探讨LncRNA BLACAT1调节miR-17-5p/DDX5轴对子宫内膜癌(EC)细胞恶性进展的影响。方法将EC细胞系Ishikawa细胞分为:NC组、si-NC组、si-BLACAT1组、si-BLACAT1+miR-NC、si-BLACAT1+miR-17-5p inhibitor组。qRT-PCR检测BLACAT1、miR-17-5p、DDX5表达水平,CCK-8法测定细胞增殖,流式细胞术检测细胞凋亡,Transwell实验检测迁移和侵袭,Western blot检测DDX5、B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)蛋白表达,荧光素酶实验检测BLACAT1、DDX5与miR-17-5p的靶向关系。结果BLACAT1、DDX5在EC细胞中表达水平升高,miR-17-5p表达降低(P<0.05),选择Ishikawa细胞进行实验。与NC组、si-NC组相比,si-BLACAT1组细胞BLACAT1、DDX5 mRNA、OD450值(24 h、48 h、72 h)、迁移和侵袭数目、Bcl-2表达降低,miR-17-5p、凋亡率、Bax表达升高(P<0.05);抑制miR-17-5p减弱了干扰BLACAT1对Ishikawa细胞增殖、迁移、侵袭的抑制,降低细胞凋亡率(P<0.05);BLACAT1靶向负调控miR-17-5p表达,miR-17-5p靶向负调控DDX5表达。结论干扰BLACAT1可通过调节miR-17-5p/DDX5轴抑制EC细胞恶性进展。

Objective To investigate the effect of LncRNA BLACAT1 on the malignant progression of endometrial carcinoma(EC)cells by regulating miR-17-5p/DDX5 axis.Methods EC cel line Ishikawa was divided into NC group,si-NC group,si-BLACAT1 group,si-BLACAT1+miR-NC,si-BLACAT1+miR-17-5p inhibitor group.The expression levels of BLACAT1,miR-17-5p and DDX5 were detected by qRT-PCR,cell proliferation was determined by CCK-8,cell apoptosis was detected by flow cytometry,migration and invasion were detected by Transwell assay.Western blot analysis was performed to detect DDx5,B-cell lymphomat-2(Bcl-2)and Bcl-2 associated X protein(Bax)protein expressions,and luciferase assay was used to detect the targeting relationship between BLACAT1,DDX5 and miR-17-5p.Results The expression levels of BLACAT1 and DDX5 were increased in EC cells,while the expression of miR-17-5p was decreased(P<0.05).Ishikawa cells were selected for the experiment.Compared with NC group and si-NC group,the BLACAT1,DDX5 mRNA,OD450 value(24 h,48 h,72 h),migration and invasion number,Bcl-2 expression were decreased in si-BLACAT1 group,while the expression of miR-17-5p,apoptosis rate and Bax were increased(P<0.05).Inhibition of miR-17-5p weakened the inhibition of interference BLACAT1 on proliferation,migration and invasion of Ishikawa cells,and decreased the apoptosis rate(P<0.05).BLACAT1 targets and negatively regulates miR-17-5p expression,and miR-17-5p targets and negatively regulates DDX5 expression.Conclusion Interference with BLACAT1 can inhibit the malignant progression of EC cells by regulating the miR-17-5p/DDX5 axis.