【摘要】 目的:观察转入野生型PTEN基因的卵巢癌细胞生物学行为变化,探索该基因对卵巢癌细胞周期的影响和抑制肿瘤侵袭的机制。方法:将野生型PTEN基因的pcDNA3.1Hygro(-)真核质粒载体转染SKOV3细胞系,潮霉素筛选稳定表达细胞株并扩增培养;采用流式细胞术检测肿瘤细胞周期变化;用MTT法检测细胞抑制率。分别用RT-PCR检测转染前后PTENmRNA表达水平变化;采用重组基底膜侵袭模型,观察转染前后细胞侵袭力的变化。结果:成功转染SKOV3并稳定表达PTEN,转染PTEN可以明显改变SKOV3细胞周期,使其阻滞于S期;明显抑制肿瘤细胞的侵袭力。结论:提示转染外源野生型PTEN基因可使SKOV3细胞周期阻滞于S期,并且通过抑制肿瘤细胞侵袭与生长,而具有明显的抑癌作用。更多还原

【Abstract】 AIM: To investigate the role of phosphatase and tensin homology deleted on chromosome(PTEN) gene in the cell cycle and invasion ability of human ovarian carcinoma SKOV3 cell line in vitro.METHODS: Human ovarian carcinoma SKOV3 cells were transfected with a eukaryotic expression plasmid vector containing PTEN gene in vitro,and then the positive cell clones were selected and amplified.MTT method was used to observe the inhibitory rate,flow cytometry was used to detect the cycle of transfected PTEN cells and apoptosis level.Western blotting analysis was used to determine PTEN gene expression.The invasiveness of transfected cells were measured quantitatively by Matrigel invasion assays(Transwell chamber).RESULTS: The expression of PTEN mRNA in SKOV3 cells increased after transfection with PTEN gene.Flow cytometry showed that the percentage of cells in S phase increased,but that in G2/M phase decreased.Invasiveness of SKOV3 was significantly decreased.CONCLUSION: The transfection of PTEN gene into SKOV3 cells can inhibit human ovarian carcinoma SKOV3 cell proliferation,invasion and induce SKOV3 cell apoptosis.更多还原