Objective To investigate the effects and possible mechanisms of miRNA-302b on "stemness" maintaining of colon cancer HT-29 cells.
Methods Human colon cancer cell line HT-29 was cultivated in vitro, and miRNA-302b mimics were transfected into HT-29 cells, while the non-transfection group (control) and negative control-mimics transfection group (NC-mimics) were taken as controls. Real-time PCR was used to detect mRNA expressions of colon cancer "stemness" markers (Ascl2, Lgr5, Oct4 and Sox2). Western blot was used to detect their protein expressions. Colony formation experiment was used to detect the colony formation ability of HT-29 cells, and stem cell sphere formation experiment was used to detect the sphere formation ability.
Results Real-time PCR showed that the mRNA expressions of "stemness" markers (Ascl2, Lgr5, Oct4 and Sox2) were up-regulated 48h after miR-302b mimics transfection into HT-29 cells (P < 0.05); Western blot showed that the protein expressions were also up-regulated (P < 0.05). Colony formation and sphere formation experiment showed that the colony and sphere formation abilities of HT-29 cells in miRNA-302b mimics transfection group were enhanced, compared with control group and NC-mimics group (P < 0.05).
Conclusion miRNA-302b enhances the colony and sphere formation abilities of HT-29 cells, through upregulating the expressions of "stemness" markers (Ascl2, Lgr5, Oct4 and Sox2), thus, miR-302b plays an important role in "stemness" maintenance of colon cancer.
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