LINC00909靶向miR-365a-5p/FGFBP1分子轴调控结肠癌细胞增殖、迁移及侵袭的实验研究

目的探讨LINC00909对结肠癌细胞增殖、迁移、侵袭的影响及可能机制。方法购买正常结肠上皮细胞NCM460,结肠癌细胞HCT8、Caco-2及DLD-1;用实时荧光定量PCR(RT-qPCR)及蛋白质印迹法(Western blotting)检测NCM460细胞及结肠癌细胞中LINC00909、miR-365a-5p及成纤维细胞生长因子结合蛋白1(FGFBP1)表达水平。将LINC00909小干扰RNA、miR-365a-5p模拟物、FGFBP1小干扰RNA分别转染HCT8细胞,用细胞计数试剂盒(CCK)-8检测细胞活力,Transwell实验检测细胞迁移及侵袭能力。双荧光素酶报告基因实验确定LINC00909与miR-365a-5p、miR-365a-5p与FGFBP1的靶向调控关系。结果与NCM460细胞比较,结肠癌细胞中LINC00909及FGFBP1表达显著升高,miR-365a-5p表达显著降低(P<0.05)。抑制LINC00909表达后,HCT8细胞活力、迁移及侵袭能力显著降低(P<0.05)。过表达miR-365a-5p后,HCT8细胞活力、迁移及侵袭能力显著降低(P<0.05)。抑制FGFBP1表达后,HCT8细胞活力、迁移及侵袭能力显著降低(P<0.05)。LINC00909靶向负调控miR-365a-5p表达,miR-365a-5p靶向负调控FGFBP1表达。抑制miR-365a-5p表达能够逆转抑制LINC00909对HCT8细胞增殖、迁移及侵袭的影响(P<0.05)。结论抑制LINC00909表达可降低结肠癌细胞增殖、迁移及侵袭能力,其机制与靶向调控miR-365a-5p/FGFBP1分子轴有关。

LINC00909 regulates the proliferation,migration and invasion of colon cancer cells by targeting miR-365a-5p/FGFBP1molecular axis

Objective To explore the effect of LINC00909 on the proliferation,migration and invasion of colorectal cancer cells and its and possible mechanism.Methods Normal colon epithelial cells NCM460,colon cancer cells HCT8,Caco-2 and DLD-1 were purchased.Real-time quantitative PCR(RT-qPCR)and Western blotting were used to detect the expression levels of LINC00909,miR-365a-5p and Fibroblast growth factor binding protein 1(FGFBP1)in NCM460 cells and colorectal cancer cells.LINC00909 small interfering RNA,miR-365a-5p mimetics,and FGFBP1small interfering RNA were transfected into HCT8 cells,respectively.Cell counting kit(CCK-8)was used to detect cell viability,transwell assays was used to detect cell migration and invasion.The dual luciferase reporter gene assays was used to confirmed the relationship between LINC00909 and miR-365a-5p,miR-365a-5p and FGFBP1.Results Compared with NCM460 cells,the expression of LINC00909 and FGFBP1in colon cancer cells was significantly increased,while the expression of miR-365a-5p was significantly reduced(P<0.05).After inhibiting the expression of LINC00909,the viability,migration and invasion ability of HCT8 cells were significantly reduced(P<0.05).After overexpression of miR-365a-5p,the viability,migration and invasion ability of HCT8 cells were significantly reduced(P<0.05).After inhibiting the expression of FGFBP1,the viability,migration and invasion ability of HCT8 cells were significantly reduced(P<0.05).LINC00909 targets and negatively regulates miR-365a-5p expression,and miR-365a-5p targets and negatively regulates FGFBP1expression.Inhibiting miR-365a-5p could reverse the effect of inhibiting LINC00909 on the proliferation,migration and invasion of HCT8 cells(P<0.05).Conclusions Inhibiting LINC00909 could reduce the proliferation,migration and invasion ability of colon cancer cells,and its mechanism is related to the targeted regulation of the molecular axis of miR-365a-5p/FGFBP1.