Effects of lncRNA TNK2-AS1 on proliferation and migration of colorectal cancer cells by regulating miR-802 expression
Objective To investigate the effect and mechanism of long chain non-coding RNA TNK2 anti-sense RNA 1(lncRNA TNK2-AS1,TNK2-AS1 for short)regulating the expression of microRNA-802(miR-802)on the proliferation,migration and invasion of colorectal cancer HT29 cells.Methods The expression of TNK2-AS1 and miR-802 in rectal cancer tissues and cells was detected by qRT-PCR;the dual luciferase reporter gene experi-ment,RNA pull down and RNA-binding protein immunoprecipitation(RIP)assay were applied to verify the target-ing relationship between TNK2-AS1 and miR-802;cells were grouped into NC group,pcDNA-NC group,pcDNA-TNK2-AS1 group,si-NC group,si-TNK2-AS1 group,si-TNK2-AS1+inhibitor NC group,and si-TNK2-AS1+miR-802 inhibitor group;Western blot was applied to detect protein expression;cell proliferation was detected by MTT and EdU methods,respectively;Transwell was applied to detect cell migration and invasion;detect the mass and volume of transplanted tumors in nude mice,as well as the expression of TNK2-AS1 and miR-802.Results Com-pared with adjacent tissues,TNK2-AS1 expression was increased(1.00±0.05,2.83±0.21 respectively,P＜0.05)in colorectal cancer tissues,and miR-802 expression was decreased(1.00±0.06,0.44±0.03 respectively,P＜0.05);Compared with normal colon epithelial cells NCM460,TNK2-AS1 expression(1.00±0.05)in colorectal cancer cell lines SW480,SW620,HCT116 and HT-29 was increased(1.64±0.19,1.37±0.36,1.48±0.30,1.92±0.25 respec-tively,all P＜0.05),miR-802 expression decreased(0.65±0.08,0.53±0.04,0.69±0.05,0.41±0.02 respectively,all P＜0.05);The double luciferase reporter gene experiment showed that TNK2-AS1 and miR-802 had a targeting re-lationship;overexpression of TNK2-AS1 promoted HT29 cell proliferation,migration,and invasion,while downreg-ulating miR-802 expression(P＜0.05);Compared with si-NC group,the expression of TNK2-AS1,proliferative abil-ity,expression of CyclinD1,cell migration,invasion number,and expression of MMP-2 and MMP-9 in HT29 cells in si-TNK2-AS1 group were obviously decreased,the expression of miR-802 was obviously increased(P＜0.05);Inhibition of miR-802 expression reverses the inhibitory effect of silencing TNK2-AS1 expression on HT29 cell prolif-eration,migration,and transplanted tumor in nude mice(P＜0.05).Conclusion TNK2-AS1 is up-regulated in colorectal cancer tissues and cells.Silencing TNK2-AS1 expression can target up-regulation of miR-802 expression to inhibit the proliferation,migration,and growth of colorectal cancer cells.
Long non-coding RNA TNK2 antisense RNA 1Micro RNA-802Colorectal cancerPro-liferationMigration
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