lncRNA CASC2对结直肠癌细胞功能及血管生成的影响

目的 研究lncRNA CASC2过表达对结直肠癌细胞增殖、迁移、血管生成及Wnt/FZD/β-catenin信号通路的影响。方法 qRT-PCR检测lncRNA CASC2在正常结肠上皮细胞(NCM460)及结直肠癌细胞株(DLD-1、HT29、SW620、HCT116、RKO、LOVO、SW480)中的表达情况;HCT116和SW480细胞建立稳定转染的细胞株,分为对照组和lncRNA CASC2过表达组;qRT-PCR验证稳转株中lncRNA CASC2水平;CCK-8实验、Transwell实验、划痕实验分别检测细胞增殖水平、迁移能力;收集对照组和lncRNA CASC2过表达组细胞上清并提取外泌体,将对照组和lncRNA CASC2过表达组细胞的外泌体分别与HUVEC细胞共孵育,3D Matrigel法检测其对血管生成的影响;Western blot测定细胞中Wnt16、FZD7、β-catenin蛋白水平。结果 lncRNA CASC2在7株结直肠癌细胞株中的表达明显低于正常结肠上皮细胞NCM460细胞。与对照组相比,lncRNA CASC2过表达后明显抑制了HCT116和SW480细胞的增殖、迁移能力(P＜0.01)。3D Matrigel法检测显示,来自SW480-CASC2组、HCT116-CASC2组细胞的外泌体显著抑制了HUVEC中管状细胞的形成(P＜0.01)。Western blot结果显示,lncRNA CASC2过表达后Wnt16、FZD7、β-catenin蛋白水平降低(P＜0.01)。结论 lncRNA CASC2在结直肠癌细胞中表达下调,lncRNA CASC2过表达可以抑制Wnt/FZD/β-catenin信号通路进而抑制结直肠癌细胞的增殖、迁移,并减少血管生成。

The effect of lncRNA CASC2 on the function and angiogenesis of colorectal cancer cells

Objective The of this study was to investigate the effect of lncRNA CASC2 overexpression on the proliferation,migration,angiogenesis and Wnt/FZD/β-catenin signaling pathway of colorectal cancer cells.Methods qRT-PCR was used to detect the expression of lncRNA CASC2 in normal colon epithelial cells(NCM460)and colorectal cancer cell lines(DLD-1,HT29,SW620,HCT116,RKO,LOVO,and SW480).HCT116 cells and SW480 cells were established stable transfection.The transfected cell lines were divided into the control group and lncRNA CASC2 overexpression group.CCK-8,Transwell,and scratch assays were used to detect the cell proliferation and migration ability.The cell supernatants collected from the control group and the lncRNA CASC2 overexpression group were used to extract exosomes,which were incubated with HUVEC cells.Their effects on angiogenesis were detected by 3D Matrigel method.The protein expression of Wnt16,FZD7 and β-catenin were determined by Western blot.Results The expression of lncRNA CASC2 in 7 colorectal cancer cell lines was significantly lower than that in normal colon epithelial NCM460 cells.Compared with the control group,the overexpression of lncRNA CASC2 significantly inhibited the proliferation,migration of HCT116 cells and SW480 cells(P＜0.01).3D Matrigel assay showed that exosomes from cells in SW480-CASC2 and HCT116-CASC2 groups significantly inhibited the formation of tubular cells in HUVEC(P＜0.01).The results of Western blot showed that the levels of Wnt16,FZD7 and β-catenin protein decreased after lncRNA CASC2 overexpression(P＜0.01).Conclusion The expression of lncRNA CASC2 is down-regulated in colorectal cancer cells.Overexpression of lncRNA CASC2 can inhibit the proliferation and migration of colorectal cancer cells by inhibiting the Wnt/FZD/β-catenin signaling pathway,and reducing angiogenesis.