PIAS3过表达对肺腺癌细胞A549增殖的影响及意义

Effect of PIAS3 over expression on proliferation in adenocarcinoma A549 cells and the significance

目的:探索活化型信号转导子和转录激活子3蛋白抑制剂(PIAS3)对肺腺癌A549细胞增殖的影响及相关意义,为进一步研究利用辐射应答基因启动子驱动的PIAS3腺相关病毒载体联合放射治疗肺腺癌提供基础。方法:利用MTT比色试验和克隆形成率实验检测转染p CMV-Sport6-PIAS3后对肺腺癌细胞A549存活和增殖的影响;利用免疫荧光染色实验检测共转染p CMVSport6-PIAS3、PRC-CMV、PRC-STAT3及PRC-STAT3-CT后对p-STAT3入核的影响。结果:PIAS3过表达导致A549细胞生长显著抑制,克隆形成能力显著降低;免疫荧光染色实验结果表明,转染p CMV-Sport6-PIAS3后,STAT3入核被显著抑制,通过共转染表达STAT3和STAT3-CT后,STAT3入核仍显著受到抑制。结论:PIAS3过表达可抑制STAT3的入核,继而阻断其功能,从而降低肺腺癌细胞A549的生长和增殖能力。

Objective： To explore the effect of PIAS3 on proliferation of lung adenocarcinoma A549 cells, in order to provide a foundation for the further study in which a adeno-associated virus （AAV） containing PIAS3 gene driven by a radiation response promoter plus radiotherapy will be used to treatment lung adenocarcinoma. Methods： The growth and proliferation of A549 cells transfected with pCMV-Sport6-PIAS3 plasmid were detected by MTT assay and cloning forming efficiency assay. The nucleus translocation of p-STAT3 was detected by immunofluorescence staining after CMV-Sport6-PIAS3 was co-transfected with PRC-CMV, PRC-STAT3 or PRC-STAT3-CT. Results：The growth and cloning form of A549 cells were significantly inhibited by PIAS3 overexpression. The result of immunofluorescence staining indicated that the nucleus translocation of p-STAT3 as significantly blocked by transfection with pCMV-Sport6-PIAS3, even after co-transfection with STAT3 or STAT3-CT. Conclusion： The overexpression of PIAS3 could inhibit the nucleus translocation of STAT3, which subsequently blocked the function of STAT3 and decreased the ability of growth and proliferation of A549 cells.