SOCS3基因在人肺腺癌细胞株A549中的表达及其对细胞增殖的影响

Expression of SOCS3 gene and its effect on cell proliferation in human lung adenocarcinoma cell line A549

目的 探讨SOCS3基因在人肺腺癌细胞株A5 49中的表达及其对细胞增殖的影响。方法 以脂质体为载体将 pEFSOCS3和 pSV2neo共转染至人肺腺癌细胞株A5 49,利用G418筛选获得阳性克隆 ;应用RT PCR和免疫细胞化学法测定转染前后细胞中SOCS3表达情况 ;采用四甲基偶氮唑兰 (MTT)显色法检测基因转染前后细胞生长情况 ;应用流式细胞仪进行细胞周期分析。结果 RT PCR和免疫细胞化学法证实转染前A5 49细胞中无SOCS3基因表达 ,转染后A5 49细胞中SOCS3基因呈稳定表达。MTT检测显示转染pEFSOCS3 的A5 49细胞生长明显受到抑制 ,抑制率为 41.0 7%。流式细胞仪检测结果显示转染 pEFSOCS3的A5 49细胞G0 /G1期细胞比例显著增高 ,S、G2 /M期细胞比例显著降低。

Objective To investigate the expression of the SOCS3 gene and its effect on proliferation of A549 cells. Methods A549 cells were cotransfected with pEFSOCS3 and pSV2neo by liposome, then G418 was used to screen the positive cells. Expression of SOCS3 mRNA and protein was detected by RT-PCR and immunocytochemistry respectively before and after transfection. MTT assay was used to detect the cell growth. Flow cytometric DNA analysis was used to determine the cell cycle. Results RT-PCR and immunocytochemistry showed that no expression of SOCS3 mRNA and protein was detected in A549 cells before transfection, but a stable expression of SOCS3 gene was observed after transfection with SOCS3 gene. Compared with control group, growth of A549 cells transfected with SOCS3 gene was significantly suppressed, with a suppressive rate of 41.07%. The cells at G 0/G 1 cell phases increased, and those at S and G 2/M phases decreased significantly after transfection. Conclusion SOCS3 protein might inhibit the proliferation of A549 cells by negatively regulating cellular signal pathways.