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目的研究转染LyGDI基因对肺腺癌A549细胞生长及放、化疗敏感性的影响。方法用脂质体介导的方法,将构建于pEGFP载体上的LyGDI及空载体pEGFP-C1转入A549细胞,用G418筛选出稳定表达株,RT-PCR法了解LyGDI基因在转染细胞中的表达情况;用平板克隆形成实验研究细胞生长情况;体外划痕试验观察对细胞侵袭及转移能力的影响;甲基偶氮唑蓝法、克隆形成实验分别研究细胞对放、化疗的敏感性。结果转染细胞中有相应融合基因表达;与转染前比较,基因转染后,阳性细胞尤其是表达LyGDI的A549细胞克隆形成能力显著降低(P<0.01),对常用化疗药物顺铂和放射线敏感度显著提高(P<0.05或<0.01)。结论建立了稳定转染细胞株;LyGDI转入抑制了A549细胞的生长能力,且增加了细胞对放、化疗的敏感性。
Objective To investigate the effects of transfected LyGDI gene on the growth and chemosensitivity of lung adenocarcinoma A549 cells. Methods LyGDI and empty vector pEGFP-C1 constructed on pEGFP vector were transfected into A549 cells by liposome-mediated method. Stable expression vector was screened by G418. The expression of LyGDI gene in transfected cells was detected by RT-PCR The cell growth was evaluated by plate clone formation assay. The in vitro scratch test was used to observe the effect on cell invasion and metastasis. Methylzidine blue assay and colony formation assay were used to study the cell sensitivity to radiotherapy and chemotherapy. Results The transfected cells showed the corresponding fusion gene expression. Compared with those before transfection, the positive cells, especially LyGDI-expressing A549 cells, had significantly lower colony-forming ability (P <0.01) Sensitivity increased significantly (P <0.05 or <0.01). Conclusions Stable transfected cell lines were established. LyGDI transfection inhibited the growth of A549 cells and increased the sensitivity of cells to radiotherapy and chemotherapy.