lncRNA-HCG11通过调控miR-144-3p/PRP11分子轴参与结直肠癌细胞增殖及转移的分子机制研究

Explore the Regulation Effect of lncRNA-HCG11 on the Proliferation,Invasion and Metastasis of Colorectal Cancer Cells and Its Molecular Mechanism

目的探讨lncRNA-HCG11对结直肠癌细胞增殖、侵袭和转移的影响作用及其分子机制。方法采用qRT-PCR检测人结直肠癌细胞和正常结直肠上皮细胞中lncRNA-HCG11的表达水平;双荧光素酶报告系统检测lncRNA-HCG11与miR-144-3p、miR-144-3p与PRP11的相互作用关系;Western blotting检测Lovo细胞中PRP11的表达水平;CCK-8法、Transwell实验分别检测结直肠癌细胞的增殖、侵袭和迁移能力。结果qRT-PCR检测结果表明lncRNA-HCG1在3株结直肠癌细胞中的表达水平均显著高于正常结直肠上皮细胞(P<0.01或P<0.001);敲降lncRNA-HCG11可显著抑制Lovo细胞的增殖、侵袭和迁移(P<0.05或P<0.01),过表达lncRNA-HCG11则得到相反的结果(P<0.05或P<0.01)。双荧光素酶报告基因实验证实lncRNA-HCG11与miR-144-3p、miR-144-3p与PRP11之间存在靶向作用关系,过表达miR-144-3p显著抑制Lovo细胞的增殖、侵袭和迁移(P<0.05或P<0.01);抑制miR-144-3p表达显著促进Lovo细胞的增殖、侵袭和迁移(P<0.05或P<0.01);回复实验表明,lncRNA-HCG11与PRP11竞争结合miR-144-3p,促进Lovo细胞的增殖、侵袭和迁移能力。结论lncRNA-HCG11作为竞争性内源RNA,与PRP11 mRNA竞争结合miR-144-3p,从而促进结直肠癌细胞Lovo增殖、侵袭和转移。

Objective To explore the regulation effect of lncRNA-HCG11 on the proliferation,invasion and metastasis of colorectal cancer cells and its molecular mechanism.Methods The expression levels of lncRNA-HCG11 in colorectal cancer cells and normal colorectal epithelial cells was detected by real-time fluorescence quantitative PCR.The interaction between lncRNA-HCG11 with miR-144-3p,and miR-144-3p with PRP11 were detected by the dual-luciferase reporter system.The expression level of PRP11 in Lovo cells was detected by Western blotting.CCK-8 assay and Transwell assay were used to detect the ability of cell proliferation,invasion and migration,respectively.Results The results of qRT-RCR showed that the expression level of lnc-HCG11 in 3 colorectal cancer cells was significantly higher than that in normal colorectal epithelial cells(P<0.01 or P<0.001).Knockdown of lncRNA-HCG11 significantly inhibited the proliferation,invasion and migration of Lovo cells,while overexpression of lncRNA-HCG11shows opposite results(P<0.05 or P<0.01).Dual luciferase reporter gene assay confirmed that lncRNA-HCG11 targeted with miR-144-3p,and miR-144-3p targeted with PRP11.Overexpression of miR-144-3p could significantly inhibit the proliferation,invasion and migration of Lovo cells(P<0.05 or P<0.01).Down-regulation of miR-144-3p expression had opposite effects on proliferation,invasion and migration of Lovo cells(P<0.05 or P<0.01).Recovery experiments showed that lncRNA-HCG11 and PRP11 competed to bind miR-144-3p and promote the proliferation,invasion and migration of Lovo cells.Conclusion lncRNA-HCG11 is a competitive endogenous RNA,which competes with PRP11 mRNA to bind miR-144-3p,thus promoting proliferation,invasion and metastasis of Lovo cells.