lncRNA-ATB促进结肠癌细胞侵袭与转移的机制研究

LncRNA-ATB promotes invasion and metastasis of colon cancer cells

摘要:

研究被转化生长因子β活化的长链非编码RNA（LncRNA-ATB）对人结肠癌细胞侵袭和转移的影响及其潜在机制。方法通过荧光实时定量聚合酶链反应检测LncRNA-ATB在结肠癌细胞系及人正常结肠上皮细胞中的表达、LncRNA-ATB shRNA 沉默效果和pcDNA3.1-LncRNA-ATB 的过表达效果，以及LncRNA-ATB 和白介素11（IL-11）mRNA 的表达；细胞侵袭和转移实验检测LncRNA-ATB 对结肠癌细胞侵袭和转移能力的影响；RNA结合蛋白免疫沉淀实验检测LncRNA与IL-11 mRNA的结合能力；酶联免疫吸附法检测结肠癌细胞上清中IL-11蛋白的表达。结果LncRNA-ATB在结肠癌细胞系中的表达较人正常上皮细胞增高；LncRNA-ATB-shRNA可有效沉默HT29 细胞中LncRNA-ATB 的表达，pcDNA3.1-LncRNA-ATB 可有效上调SW620细胞中LncRNA-ATB的表达；LncRNA-ATB可促进结肠癌细胞的侵袭和转移能力。LncRNAATB可与IL-11 mRNA结合并调控其表达，增强其mRNA稳定性，促进其分泌。结论LncRNA-ATB可通过介导炎症因子IL-11 的分泌，在结肠癌细胞的侵袭和转移过程中发挥重要作用。

Abstract:

To investigate the effect of long non-coding RNA activated by TGF-β(LncRNAATB) on the migration and invasion of human colorectal cancer cells and the underlying mechanism. Methods The efficiency of LncRNA-ATB shRNA and pcDNA3.1-LncRNA-ATB, and the expressions of LncRNA-ATB and IL-11 mRNA were detected by quantitative real time PCR (qRT-PCR). The effect of LncRNA-ATB on migration and invasion abilities of colorectal cells were analyzed by cell migration and cell invasion assays.The binding ability of LncRNA-ATB and IL-11 mRNA was analyzed by RNA immunoprecipitation (RIP). The expression of IL-11 protein was analyzed by ELISA. Results The expression of LncRNA-ATB in the colorectal cells was higher than that in the normal human colon epithelial cells. LncRNA-ATB expression in the HT29 cells could be effectively silenced by LncRNA-ATB shRNA and LncRNA-ATB expression in the SW620 cells could be efficiently upregulated by pcDNA3.1-LncRNA-ATB. LncRNA-ATB could significantly promote the migration and invasion of colorectal cells. Binding to IL-11 mRNA, LncRNA-ATB could upregulate IL-11mRNA expression, enhance the mRNA stability of IL-11 and promote its secretion in colorectal cancer cells. Conclusions LncRNA-ATB may play an important role in the migration and invasion of colorectal cells by enhancing the secretion of IL-11.