lncRNA BLACAT1靶向LEMD1促进结直肠癌细胞的增殖和转移

LncRNA BLACAT1 targeting LEMD1 promotes the proliferation and metastasis of colorectal cancer cells

目的研究长链非编码RNA(lncRNA)BLACAT1在结直肠癌细胞发生发展过程的作用机制。方法starBase分析TCGA数据库中lncRNA BLACAT1分别在结直肠癌组织和癌旁组织中的表达水平,采用qPCR分别检测10例结直肠癌患者癌组织和癌旁组织中lncRNA BLACAT1的表达水平;检测结直肠癌细胞系SW620、SW480、HT-29、LoVo、HCT116和正常结直肠上皮细胞FHC中lncRNA BLACAT1的mRNA表达水平,筛选lncRNA BLACAT1高表达细胞系;敲低lncRNA BLACAT1验证对高表达细胞系增殖、迁移和侵袭的影响;starBase在线预测与lncRNA BLACAT1靶向作用的基因,qPCR和Western blot实验验证结果;starBase分析lncRNA BLACAT1与作用基因相关性,双荧光素酶报告基因实验验证lncRNA BLACAT1的靶基因;检测lncRNA BLACAT1作用靶基因对结直肠癌细胞的增殖、迁移和侵袭的影响。结果TCGA数据库分析和结直肠癌患者检测结果均表明癌组织中lncRNA BLACAT1表达明显高于癌旁组织(P<0.001);结直肠癌细胞系SW620中lncRNA BLACAT1的mRNA表达水平最高(P<0.001);敲低lncRNA BLACAT1能够抑制SW620细胞的增殖(P<0.001)、迁移(P<0.001)和侵袭能力(P<0.001);qPCR和Western blot实验及数据库分析结果表明lncRNA BLACAT1与LEMD1的表达存在正相关关系(r=0.778,P<0.001),双荧光素酶报告基因实验证明LEMD1是lncRNA BLACAT1的靶基因;lncRNA BLACAT1上调LEMD1的表达,促进SW620细胞的增殖(P<0.001)、迁移(P<0.001)和侵袭(P<0.001)。结论lncRNA BLACAT1靶向LEMD1促进结直肠癌细胞SW620的增殖和转移。

Objective The mechanism of long-chain non-coding RNA(lncRNA)BLACAT1 was studied in the occurrence and development of colorectal cancer cells.Methods StarBase was used to analyze the expression of lncRNA BLACAT1 in colorectal cancer tissues and adjacent tissues by the TCGA database.qRT-PCR was used to detect the expression of lncRNA BLACAT1 in tumor and adjacent tissues of 10 colorectal cancer patients.The expression of lncRNA BLACAT1 at the mRNA level was detected in normal colorectal epithelial FHC cells and several colorectal cancer cell lines of SW620,SW480,HT-29,LoVo and HCT116.The high expression of lncRNA BLACAT1 was screened in these cell lines.The effects of lncRNA BLACAT1 knockdown on proliferation,migration and invasion were verified in colorectal cancer cell lines.Targets of lncRNA BLACAT1 predicted by starBase online database were verified by qPCR and western blot.The correlation between lncRNA BLACAT1 and its target gene was validated by dual luciferase reporter gene.Finally,effects of target genes of lncRNA BLACAT1 on cell proliferation,migration and invasion of colorectal cancer cells were detected.Results Both database analysis and colorectal cancer patient detection results showed that the expression of lncRNA BLACAT1 in tumor tissues was significantly higher than that in adjacent tissues(P<0.001).The expression of lncRNA BLACAT1 mRNA was the highest in colorectal cancer SW620 cells(45.180±0.892,t=110.6,P<0.001).Knockdown of lncRNA BLACAT1 could inhibit the proliferation(P<0.001),migration(P<0.001)and invasion(P<0.001)ability of SW620 cells.The results of qPCR and Western blot and database analysis showed that there was a positive correlation between the expression of lncRNA BLACAT1 and LEMD1(r=0.778,P<0.001).The dual luciferase reporter gene experiment proved that LEMD1 was the target gene of lncRNA BLACAT1.lncRNA BLACAT1 up-regulated the expression of LEMD1 and promoted the proliferation(P<0.001),migration(P<0.001)and invasion(P<0.001)of SW620 cells.Conclusion lncRNA BLACAT1 targeting LEMD1 promotes the proliferation and metastasis of colorectal cancer SW620 cells.