细胞融合通过PI3K-AKT通路降低黑色素瘤细胞增殖能力

Cell fusion reduces melanoma cell proliferation through PI3K-AKT pathway

目的研究细胞融合在黑色素瘤细胞增殖中的作用及其分子机制。方法用聚乙二醇细胞融合方法融合黑色素瘤细胞,通过流式分选获得稳定的融合细胞。体外培养及细胞计数检测融合细胞的增殖速度。Affymatrix基因芯片分析差异表达的基因,IPA软件进行基因功能分析。Western blot检测细胞蛋白。结果 PEG化学融合法成功获得稳定的黑色素瘤融合细胞,其体外增殖速度明显减慢(P<0.01)。黑色素瘤融合细胞与增殖相关的PI3K-AKT通路明显下调(P<0.01)。融合细胞phospho-S6蛋白表达明显降低。结论细胞融合通过PI3K-AKT通路降低黑色素瘤细胞增殖能力。

Objective To investigate the effect of cancer cell fusion in melanoma proliferation and the underlying mechanism.Methods Stable melanoma fusion cells were obtained by the polyethylene glycol（PEG） fusion method and isolated by the fluorescence activated cell sorting（FACS）.Cell proliferation was examined by cell culture in vitro and the cell number was counted daily.Differentially expressed genes between the stable melanoma fusion hybrids and the parental cells were detected by the Affymatrix gene chip following the gene function analysis by Ingenuity Pathways Analysis（IPA）.Protein expression was analyzed by Western blot.Results Stable melanoma fusion cells were successfully obtained by the PEG fusion method.The proliferation rate of fusion hybrids in vitro was significantly decreased.PI3K-AKT pathway related to cell proliferation was significantly down-regulated in the melanoma fusion hybrids.Moreover,the expression of phospho-S6 in the fusion hybrids significantly decreased.Conclusions Cell fusion reduces melanoma cell proliferation through the PI3K-AKT pathway.