Effect of miR-27a on cell proliferation and apoptosis in human melanoma cell line WM239 / 国际肿瘤学杂志

ABSTRACT

Objective To investigate the effect of miR-27a mimic and inhibitor on proliferation and apoptosis in melanoma cell line WM239. Methods The miR-27a mimic,inhibitor and its negative control were transfected into WM239 cells. The transfection efficiency was evaluated by fluorescence microscope. The expres-sion of miR-27a was detected by real-time fluorescent quantitative PCR. The proliferation of cells was detected by MTT. The cell apoptosis and cell cycle were analyzed by flow cytometry. Results The transfection efficiency in WM239 cells was 80% to 90%. The expression of miR-27a was markedly up-regulated in miR-27a mimic group (2-△△CT value is 26. 98 ± 0. 01),with statistically significant difference(t= -1 123. 67,P=0. 00);and the miR-27a inhibitor group showed lower expression of miR-27a(2-△△CT value is 0. 96 ± 0. 02),there was no statisti-cally significant difference compared with normal control group(t=0. 04,P=0. 06). The proliferation of cells was obviously inhibited in miR-27a mimic group,and there was statistically significant difference compared with normal control group[absorbance of 72 h(0. 45 ± 0. 02)∶(0. 72 ± 0. 01),F=129. 56,P﹤0. 05]. The percent-age of WM239 cells in G0-G1 phase was increased[(74. 83 ± 1. 46)∶(63. 73 ± 1. 25),F=30. 33,P﹤0. 05], and the percentage of WM239 cells in S phase and G2-M phase were decreased[(21. 33 ± 1. 75)∶(27. 50 ± 1. 25),F=14. 98,P﹤0. 05;(3. 90 ± 1. 31)∶(8. 80 ± 2. 10),F=3. 66,P﹤0. 05]. The apoptosis rate of cells was significantly increased in miR-27a mimic group compared with normal group[(29. 67 ± 0. 91)%∶(1. 44 ± 0. 85)%,F=530. 90,P﹤0. 01],but the inhibitor group had no obvious effect on cell cycle and cell apoptosis. Conclusion MiR-27a can suppress melanoma cell proliferation and act as a tumor suppressor gene,which is rel-evant to induce cell apoptosis and block cell cycle in G0-G1 phase.