Abstract:
Objective To investigate the effect of antisense oligonucleotides(ASOs) against miR-21 expression on the growth of human colon carcinoma cells line HCT116 in vivo.
Methods The nude mouse model of human colon carcinoma cell line HCT116 was established. The p-miR-21-ASOs plasmid and p-Cont plasmid (100μg/body) were inoculated subcutaneously into xenograft tumor, and the tumor growth was observed. HE staining was used to observe the morphological changes of tumor; immunofluorescence technique was used to detect tumor core antigen Ki-67 protein expression; the expression levels of mature miR-21, as well as cyclin-dependent kinase 2 (CDK2), CDK3, CDK4 and CDK6 were detected by real-time fluorescent quantitative PCR. And the levels of p-Akt, p-ERK1/2, total Akt and ERK1/2 protein were analyzed by Western blot.
Results Compared with p-Cont group, the tumor cells in the p-miR-21-ASOs group had a slower growth rate, and the tumor quality was significantly decreased (P < 0.05); moreover, the expression level of Ki-67 was decreased significantly (P=0.0074); the expression level of miR-21, CDK2, CDK3, CDK4 and CDK6 were significantly decreased (all P < 0.05); the relative expression levels of p-Akt and p-ERK were significantly down-regulated (P < 0.05).
Conclusion The down-regulation of miR-21 expression by ASOs could significantly inhibit the growth of human colon cancer cells in vivo.
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