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 ...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。展开更多
Background and aims:Raf kinase inhibitor protein(RKIP)is an important member of the phos phatid ylethanolamine-binding protein family.This study investigated the inhibitory effect of RKIP on the malignant biological b...Background and aims:Raf kinase inhibitor protein(RKIP)is an important member of the phos phatid ylethanolamine-binding protein family.This study investigated the inhibitory effect of RKIP on the malignant biological behavior of liver cancer cells in vivo and in vitro.Methods:An RKIP gene expression vector was constructed using the pcDNA3.1 vector and transfected into human hepatoma cell line HepG2 to overexpress the RKIP gene.Growth,proliferation,the cell cycle,apoptosis,migra-tion,and invasion of the cell line were analyzed.For in vivo experiments,the vector was transfected into tumor tissue of hepatoma-bearing animals and then tumor growth was analyzed and compared with the control to assess its anti-tumor effect.Results:Compared with the control group,cell growth in the RKIP gene transfection group(HEP-RKIP)was significantly slower.The average colony formation rate of the HEP-RKIP group was significantly lower than that of the other two groups(p<0.05).The proportion of HEP-RKIP cells in G0/G1 phase was significantly higher than that of the control group(p<0.05),while the proportion of cells in G2/M phase was significantly lower than that in the control group(p<0.05).The apoptosis rate of HEP-RKIP cells was approximately 6.4%,which was significantly higher than that of the control group(p<0.05).Migration of HEP-RKIP cells was significantly slower than that of the other two groups(p<0.05).In an inhibition experiment of the liver tumor animal model,the tumor inhibition rate of the RKIP in vivo transfection group was significantly higher than that of each control group(p<0.05).Conclusion:RKIP inhibits malignant biological behavior of liver cancer cells in vitro and has a tumor inhibitory effect in vivo,which may be a potential target for gene therapy of liver cancer.展开更多
文摘目的探讨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。
文摘Background and aims:Raf kinase inhibitor protein(RKIP)is an important member of the phos phatid ylethanolamine-binding protein family.This study investigated the inhibitory effect of RKIP on the malignant biological behavior of liver cancer cells in vivo and in vitro.Methods:An RKIP gene expression vector was constructed using the pcDNA3.1 vector and transfected into human hepatoma cell line HepG2 to overexpress the RKIP gene.Growth,proliferation,the cell cycle,apoptosis,migra-tion,and invasion of the cell line were analyzed.For in vivo experiments,the vector was transfected into tumor tissue of hepatoma-bearing animals and then tumor growth was analyzed and compared with the control to assess its anti-tumor effect.Results:Compared with the control group,cell growth in the RKIP gene transfection group(HEP-RKIP)was significantly slower.The average colony formation rate of the HEP-RKIP group was significantly lower than that of the other two groups(p<0.05).The proportion of HEP-RKIP cells in G0/G1 phase was significantly higher than that of the control group(p<0.05),while the proportion of cells in G2/M phase was significantly lower than that in the control group(p<0.05).The apoptosis rate of HEP-RKIP cells was approximately 6.4%,which was significantly higher than that of the control group(p<0.05).Migration of HEP-RKIP cells was significantly slower than that of the other two groups(p<0.05).In an inhibition experiment of the liver tumor animal model,the tumor inhibition rate of the RKIP in vivo transfection group was significantly higher than that of each control group(p<0.05).Conclusion:RKIP inhibits malignant biological behavior of liver cancer cells in vitro and has a tumor inhibitory effect in vivo,which may be a potential target for gene therapy of liver cancer.
