RGS4表达上调对人恶性黑素瘤细胞的M14增殖和迁移的影响

Up-regulation of RGS4 inhibits the proliferation and migration of human malignant melanoma cells in vitro

目的:明确体外RGS4基因表达上调对恶性黑素瘤M14细胞中增殖及迁移的影响。方法:体外培养M14细胞,分为对照组(转染pc DNA3.1空质粒)和实验组(转染pc DNA3.1-RGS4质粒)。RT-PCR和Western blot检测转染后RGS4 mRNA和RGS4蛋白水平的表达,CCK-8和克隆形成实验检测RGS4上调对恶性黑素瘤M14细胞增殖的影响,划痕实验检测RGS4高表达对M14细胞迁移的影响。结果:实验组细胞中RGS4 mRNA表达量是对照组的3.25×10~3倍(P<0.05),蛋白表达量是对照组的6.73倍(P<0.05)。实验组M14细胞OD值为1.33±0.07,低于对照组的1.61±0.11(P<0.05),实验组M14细胞克隆数为34±5.53,低于对照组的56±7.68(P<0.05)。实验组M14细胞迁移能力低于对照组。结论:RGS4高表达能够抑制体外恶性黑素瘤细胞的增殖和迁移,RGS4在恶性黑素瘤的发生、发展中可能发挥重要作用。

Objective：To determine the influence of pc DNA3.1-RGS4 vector up-regulating RGS4 expression on the proliferation and migration of human malignant melanoma cells in vitro.Methods：The cells were cultured in vitro,then divided into the control group（pc DNA3.1）and experimental group（pc DNA3.1-RGS4）.The expression of RGS4 mRNA and protein in M14 cell was detected by RT-PCR and Western blot.The proliferation of M14 cell was detected by CCK-8 and colon formation assay.The migration of M14 cell was detected by wound healing assay.Results：The levels of RGS4 mRNA and protein in the experimental group were 3.25×10~3and 6.73 times more than that in the control group（P_s0.05）.The OD scores and clone number of M14 cells in the experimental group were 1.33±0.07 and 34±5.53,which was lower than that in the control group（1.61±0.11 and 56±7.68）（P_s0.05）.M14 cellular migration ability in the experimental group was weaker than that in the control group.Conclusion：RGS4 up-regulation inhibits the M14 cell proliferation and migration in malignant melanoma.RGS4 may play an important role in the development of malignant melanoma.