lncRNA TDRG1通过调节miR-101-3p促进结直肠癌细胞增殖、侵袭和迁移

lncRNA TDRG1 promotes proliferation,invasion and migration of colorectal cancer cells by regulating miR-101-3p

目的:研究长链非编码RNA(lncRNA)睾丸发育相关基因1(TDRG1)对结直肠癌(CRC)细胞增殖、侵袭和迁移的影响,并探讨其作用机制是否与微小RNA-1010-3p(miR-101-3p)相互作用有关。方法:收集40对CRC组织及相应的癌旁组织。采用RT-qPCR方法检测TDRG1和miR-101-3p在CRC组织及CRC细胞系中的表达。采用CCK-8法、集落形成实验和Transwell法评价TDRG1的体外功能。采用生物信息学和双萤光素酶报告分析方法探讨TDRG1和miR-101-3p之间关系。将shRNA和miRNA抑制剂转染CRC细胞,探讨其作用机制。结果:与匹配的癌旁组织相比,在CRC组织中观察到TDRG1的表达水平显著增加(P<0.05),而miR-101-3p的表达水平显著降低(P<0.05)。Pearson相关分析显示,CRC组织中TDRG1与miR-101-3p呈负相关(r=0.624,P<0.01)。使用萤光素酶测定法确认了miR-101-3p作为TDRG1的靶点结合。与人正常结肠细胞NCM460相比,TDRG1在CRC细胞系(HT-29、SW480和LoVo)中明显上调(P<0.05)。敲减TDRG1的表达抑制了SW480细胞的活力和集落能力(P<0.05),并且迁移和侵袭细胞数量显著减少(P<0.05),而转染miR-101-3p抑制剂可减轻敲减TDRG1表达引起的改变。结论:TDRG1通过调节miR-101-3p促进CRC细胞的增殖、侵袭和迁移,提示TDRG1可能是CRC治疗的一个潜在靶点。

AIM:To investigate the effect of long noncoding RNA(lncRNA)testis development-related gene1(TDRG1)on proliferation,invasion and migration of colorectal cancer(CRC)cells,and to explore whether its mecha-nism is related to the interaction of microRNA-101-3 p(miR-101-3 p).METHODS:A total of 40 paired CRC tissues and corresponding adjacent tissues were collected.RT-qPCR was used to detect the expression levels of TDRG1 and miR-101-3 p in the CRC tissues and the cells.CCK-8,colony formation and Transwell assays were performed to evaluate the func-tions of TDRG1 in vitro.Bioinformatic analysis and luciferase report analysis were used to explore the relationship between TDRG1 and miR-101-3 p.shRNA and miRNA inhibitor were transfected into CRC cells to explore the underlying mecha-nisms.RESULTS:A significant increased expression level of TDRG1 and a significant decreased expression level of miR-101-3 p were observed in the CRC tissues,compared with matched tissues.Pearson correlation analysis showed that the expression of miR-101-3 p was negatively correlated with the expression of TDRG1(r=0.624,P<0.01).Binding of TDRG1 and miR-101-3 p was confirmed using luciferase assays.TDRG1 was up-regulated markedly in the CRC cell linesHT-29,SW480 and LoVo,in comparison with that in normal colorectal cellsNCM460(P<0.05).Knock-down of TDRG1 expression inhibited the viability and colony formation ability of SW480 cells(P<0.05),and the numbers of mi-gration and invasion cells were decreased significantly(P<0.05).Transfection of miR-101-3 p inhibitor mitigated the alter-ations induced by TDRG1 expression knock-down.CONCLUSION:TDRG1 promotes CRC cell proliferation,invasion and migration by miR-101-3 p signaling,suggesting that TDRG1 is a potential therapeutic target in CRC treatment.