circCSPP1靶向miR-495-3p调控结直肠癌细胞的增殖和糖酵解

Effects of circCSPP1 on the proliferation and glycolysis of colorectal cancer cells

目的研究环状RNA中心体/纺锤体相关蛋白基因(circCSPP1)在结直肠癌细胞增殖和糖酵解中的作用,并探索其潜在机制。方法采用实时荧光定量PCR(RT-qPCR)检测circCSPP1和miR-495-3p在结直肠癌组织、癌旁组织中的表达。将si-NC、si-circCSPP1、miR-NC、miR-495-3p mimics、si-circCSPP1+anti-miR-NC、si-circCSPP1+anti-miR-495-3p分别转染HCT116细胞,细胞计数试剂盒(CCK-8)检测细胞活力,试剂盒检测细胞中葡萄糖消耗和乳酸水平。双荧光素酶报告基因实验和RT-qPCR确定circCSPP1与miR-495-3p之间相互作用。结果与癌旁组织比较,结直肠癌组织中circCSPP1表达显著升高,miR-495-3p表达显著降低(P<0.05)。抑制circCSPP1或过表达miR-495-3p后,HCT116细胞活力显著降低,葡糖糖消耗和乳酸产生水平显著降低(P<0.05)。抑制miR-495-3p表达可逆转抑制circCSPP1对HCT116细胞活力和糖酵解反应的影响(P<0.05)。miR-495-3p是circCSPP1的靶基因,circCSPP1负调控miR-495-3p表达。结论抑制circCSPP1能够抑制胃癌细胞的增殖和糖酵解反应,其机制与靶向调控miR-495-3p有关。

Objective To investigate the effects of circular RNA centrosome/spindle pole-associated protein 1 gene(circCSPP1)on the proliferation and glycolysis of colorectal cancer cells,and to explore its potential action mechanism.Methods The expression levels of circCSPP1 and miR-495-3p in colorectal cancer tissues and adjacent tissues were detect by real-time fluorescence quantitative PCR(RT-qPCR).And si-NC,si-circCSPP1,miR-NC,miR-495-3p mimics,si-circCSPP1+anti-miR-NC,si-circCSPP1+anti-miR-495-3p were transfected into HCT116 cells,respectively.The cell viability was analyzed by cell countingkit(CCK-8),and the levels of lactic acid and glucose consumption were detected by kit.The interation between circCSPP1 and miR-495-3p was determined by dual luciferase assay and RT-qPCR.Results Compared with those in adjacent tissues,the expression levels of circCSPP1 in colorectal cancer tissues were significantly increased,whereas the expression levels of miR-495-3p were significantly decreased(P<0.05).After the inhibition of circCSPP1 or the overexpression of miR-495-3p,the viability of HCT116 cells was significantly decreased,and glucose consumption and lactate production levels were significantly decreased(P<0.05).To inhibit the expression of miR-495-3p could reverse the effects of circCSPP1inhibition on the viability and glycolysis of HCT116 cells(P<0.05).Moreover miR-495-3p was the target gene of circCSPP1,and circCSPP1 negatively regulated miR-495-3p expression.Conclusion Inhibiting circCSPP1 can inhibit the proliferation and glycolysis of gastric cancer cells,and its action mechanism is related to the targeted regulation of miR-495-3p.