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目的探讨BRAF~(V600E)突变基因在黑素瘤发展过程中的作用。方法用构建成功的质粒载体转染人黑素瘤细胞株A375,并建立动物模型来检测其在体内的干扰作用。分别采用RT-PCR和蛋白质印迹法检测体内外BRAF基因的mRNA和蛋白表达。结果特异性短发卡RNA在体内外均可抑制人黑素瘤细胞BRAF基因mRNA和蛋白的表达,使其蛋白表达量分别减少62%和90%。在动物模型中使黑素瘤细胞的成瘤性下降,可以抑制肿瘤增长缓慢,抑瘤率达到62%。结论BRAF~(V600E)突变基因在黑素瘤的发展中有一定的作用。
Objective To investigate the role of BRAF ~ (V600E) mutation in the development of melanoma. Methods Human melanoma cell line A375 was transfected with the constructed plasmid vector and the animal model was established to detect the interference effect in vivo. The mRNA and protein expression of BRAF gene in vivo and in vitro were detected by RT-PCR and Western blot respectively. Results The specific short hairpin RNA could inhibit the expression of BRAF mRNA and protein in human melanoma cells both in vitro and in vivo, and decrease the expression of BRAF mRNA by 62% and 90% respectively. In animal models, the tumorigenicity of melanoma cells is reduced, which can inhibit the slow growth of the tumor, with a tumor inhibition rate of 62%. Conclusion The BRAF ~ (V600E) mutant gene has a certain role in the development of melanoma.