circDUSP16调节miR-126-5p/SHH轴对结直肠癌细胞增殖、凋亡和上皮间质转化的影响

Impacts of circDUSP16 on the proliferation,apoptosis and epithelial-mesenchymal transition of colorectal cancer cells by regulating the miR-126-5p/SHH axis

目的:探究circDUSP16对结直肠癌(CRC)细胞增殖、凋亡和上皮间质转化(EMT)的影响及其作用机制。方法:qRT-PCR检测circDUSP16、miR-126-5p、SHH表达水平,CCK-8法测定细胞增殖,流式细胞术检测细胞凋亡,Western blot检测SHH、PCNA、Bcl-2、Bax、E-cadherin、N-cadherin蛋白表达,荧光素酶实验检测circDUSP16和miR-126-5p/SHH的靶向关系。结果:沉默circDUSP16组SW480细胞circDUSP16、SHH mRNA表达、OD450值(24 h、48 h、72 h)、Bcl-2和N-cadherin蛋白表达降低,miR-126-5p、凋亡率、Bax、E-cadherin蛋白水平升高(P<0.05);下调miR-126-5p减弱了沉默circDUSP16对SW480细胞的增殖、EMT发生的抑制,降低了细胞的凋亡能力;circDUSP16靶向负调控miR-126-5p表达,miR-126-5p靶向负调控SHH表达。结论:沉默circDUSP16可能通过上调miR-126-5p并下调SHH抑制SW480细胞增殖和EMT发生,促进凋亡。

Objective:To investigate the impacts of circDUSP16 on proliferation,apoptosis and epithelial-mesenchymal transition(EMT)of colorectal cancer(CRC)cells and its mechanism.Methods:qRT-PCR was applied to detect the expression levels of circDUSP16,miR-126-5p,and SHH,CCK-8 assay was applied to measure cell proliferation,flow cytometry was applied to detect apoptosis,Western blot was applied to detect the protein expression of SHH,Bcl-2,Bax,E-cadherin,N-cadherin),and Luciferase assay was applied to detect the targeting relationship between circDUSP16 and miR-126-5p/SHH.Results:The silencing circDUSP16,SHH mRNA expression,OD450 value(24 h,48 h,72 h),the protein expression of Bcl-2 and N-cadherin decreased,miR-126-5p,apoptosis rate,and the protein levels of Bax and E-cadherin increased(P<0.05);down-regulation of miR-126-5p attenuated the inhibition of the proliferation and EMT occurrence of SW480 cells by silencing circDUSP16,and decreased the apoptosis ability of cells;circDUSP16 targeted and negatively regulated miR-126-5p expression,and miR-126-5p targeted and negatively regulated SHH expression.Conclusion:Silencing circDUSP16 may inhibit the proliferation and EMT,and promote apoptosis of SW480 cells by up-regulating miR-126-5p and down-regulating SHH.