RKIP基因对胃癌细胞生物学行为的影响

Effect of RKIP on biological characters of gastric carcinoma cells

目的探讨RKIP基因对胃癌细胞株SGC7901恶性生物学行为的影响。方法采用脂质体转染技术,将RKIP真核表达重组质粒pc DNA3.1（＋）/RKIP和空载体质粒分别导入胃癌细胞系SGC7901,定量-反转录-PCR（q-RT-PCR）和Western blot分别检测RKIP m RNA及蛋白质表达,构建RKIP基因稳定表达的胃癌细胞系SGC7901。借助细胞生长曲线及集落形成实验观察胃癌细胞增殖的变化;划痕试验及Transwell实验检测胃癌细胞迁移能力和侵袭力的变化;流式细胞术检测RKIP表达对胃癌细胞的细胞周期和凋亡率的影响;并建立人胃癌细胞裸鼠皮下移植瘤模型,观察RKIP对胃癌细胞增殖和转移能力的影响。结果转染RKIP基因的SGC7901细胞系RKIP m RNA和蛋白质表达量显著增加。转染组SGC7901细胞较对照组生长速度减慢,集落形成率显著下降,流式细胞术分析发现,RKIP延缓细胞由G0~G1期进入S期,诱导细胞凋亡,裸鼠接种实验显示,RKIP基因可使裸鼠成瘤平均时间延长,生成移植瘤体积显著减小。结论 RKIP与胃癌的增殖、凋亡和侵袭等生物学行为密切相关,RKIP基因重表达有助于SGC7901的恶性表型逆转,是肿瘤治疗的候选有效靶点。

[ Objective ] To determine the effects of RKIP gene on biological charateristics of gastric carcinoma cell line SGC7901. [Methods] The RKIP gene eukaryotie expression vector pcDNA3.1（＋）-RKIP was transfected in- to SGC7901 cells by lipid media transfection. Cells untransfected SGC7901 cells and those transfected with pcD- NA3.1 （＋） plasmid were used as controls. The expression levels of RKIP mRNA and protein in SGC7901 cells were detected by q-RT-PCR and Westem blotting analysis, respectively. The proliferative activity of SGC7901 cells were demonstrated by drawing growth curve and soft agar assay. The invasion ability of the cells was detected by scratch as- says and transwellassay. Cell cycle distribution and the apoptosis were determined by flow cytometrie analysis. Tumor formation in nude mice was used to assess the tumorigenic characteristics in vivo. [Results] The mRNA and protein levels of RKIP in the transfected cell lines were significantly higher than those in controls. Compared with vector transfected and untansfected SGC7901 cells, RKIP gene significantly inhibited the cell proliferation and invasion, pro- moted the cell apoptosis, and decreased tumorigenicity in nude mice. Flow cytometrie data showed that more RKIP transfected cells went into phase GO-G1 than controls. [ Conclusions] These data suggest that RKIP gene may play an important role in gastric cancer cell proliferation, invasion and apoptosis; expression of RKIP gene could favor the malignant ohenotvoe revision of GC cells. And RKIP may serve as an effective target for cancer ~ene theraov.