Mechanism of lncRNA LINC01612 promoting malignant growth of esophageal squamous cell carcinoma cells
Objective To investigate the effect of lncRNA LINC01612(LINC01612)in the biological function of esophageal squamous cell carcinoma(ESCC)cells and its mechanism.Methods Tumor tissues and adjacent tissues from 42 ESCC patients were collected,and the relative expression levels of LINC01612 in ESCC tissues,adjacent tissues,ESCC cell lines and normal esophageal epithelial cells(Het-1 A cell)were detected by RT-qPCR,respectively.KYSE30 cells were divided into the control group(without transfection),the shRNA-NC group(transfected with shRNA-NC),the sh-LINC01612 group(transfected with LINC01612 shRNA),the Vector group(transfected with vector),the ALKBH5 group(transfected with ALKBH5 overexpression vector),the IGF2BP1 group(transfected with IGF2BP1 overexpression vector)and ALKBH5+LINC01612 group(co-transfected with ALKBH5 and LINC01612 overexpression vectors).Methylated RNA immunoprecipitation(MeRIP)assay was used to evaluate the m6A modification level of LINC01612;RIP analysis was used to evaluate the binding of m6A modified LINC01612 to ALKBH5 or IGF2BP1;actinomycin D experiment was used to detect the stability of LINC01612;CCK-8 assay and Transwell assay were used to detect the cell proliferation and invasion abilities;and the flow cytometry was used to detect the cell apoptosis rate.Results The expression of LINC01612 was significantly upregulated in the ESCC tissues compared with the adjacent tissues(P＜0.01).Moreover,the expression levels of LINC01612 in ESCC cells were significantly higher than that in human normal esophageal epithelial cells of Het-1A(P＜0.01).Silencing LINC01612 significantly inhibited the proliferation and invasion of KYSE30 cells,induced cell apoptosis and increased Caspase-3 activity(P＜0.05).ALKBH5 abrogated m6A modification of LINC01612 by binding to LINC01612,thereby downregulating the expression of LINC01612.Overexpression of ALKBH5 inhibited cell proliferation and invasion,and induced cell apoptosis(P＜0.01),while overexpression of LINC01612 counteracted the effects of ALKBH5 overexpression on KYSE30 cells(P＜0.05).RIP analysis showed that IGF2BP1 enhanced the stability of LINC01612 and promoted its expression by recognizing and binding LINC016122 modified by m6A.Conclusion LINC01612 was significantly upregulated in ESCC.ALKBH5-m6A-IGF2BP1 mediated LINC01612 upregulation in an m6A dependent manner,thereby promoting the proliferation and invasion of ESCC cell,and inhibiting cell apoptosis.
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维普
