Objective： The purpose of the study is to investigate the effects of up-regulation of Raf kinase inhibitor protein （RKIP） on the chemosensitivity of cervical cancer Hela cells. Methods： Eukaryotic expression plasm...Objective： The purpose of the study is to investigate the effects of up-regulation of Raf kinase inhibitor protein （RKIP） on the chemosensitivity of cervical cancer Hela cells. Methods： Eukaryotic expression plasmid pcDNA3.1（±）-ssRKIP containing human overall length RKIPcDNA was transfected into cervical cancer Hela cell by lipofectin assay, establishing a stable cell line containing a target gene by G418. Expression of RKIP in Hela cells was measured by Western blot analysis. After treatment with cisplatin of different concentrations and intervals of time, the effect of RKIP on the proliferation of Hela cells was evaluated by MTT method. The flow cytometry was used to investigate whether the RKIP could inhibit apoptosis in Hela cells induced by cisplatin. Results： The expression of RKIP in Hela cells transfected with pcDNA3.1-ssRKIP was increased obviously. After different concentrations of cisplatin treatment cells for 24, 48 and 72 h, the growth inhibition rate in Hela cells transfected with pcDNA3.1-ssRKIP was significantly higher than in control cells （P 〈 0.05）. With 5 pg/mL cisplatin treatment for 24 h, pcDNA3.1-ssRKIP-transfected Hela cells had an obviously higher percentage of apoptosis （23.2 ± 0.24）% than non-transfected cells （12.4 ± 0.31）% and empty vector-transfected cells （13.4 ± 0.47）%. Without treatment of cisplatin, the percentage of apoptosis for Hela cells transfected with pcDNA3.1-ssRKIP was （5.7 ± 0.12）%, which was still higher than those of the non-transfected cells （2.9 ± 0.21）% and empty vector-transfected cells （3 ± 0.08）%. Conclusion： Higher expres- sion of RKIP gene can improve chemosensitivitv of cervical cancer Hela cells to cisplatin.展开更多
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
目的:探讨RKIP在Ⅱ期结直肠癌中的表达及其与患者预后的关系。方法:收集130例Ⅱ期结直肠癌患者的术后组织标本,采用免疫组织化学SP法检测RKIP在原发肿瘤组织及癌旁组织中的表达,结合患者临床病理特征分析RKIP的表达水平与Ⅱ期结直肠癌...目的:探讨RKIP在Ⅱ期结直肠癌中的表达及其与患者预后的关系。方法:收集130例Ⅱ期结直肠癌患者的术后组织标本,采用免疫组织化学SP法检测RKIP在原发肿瘤组织及癌旁组织中的表达,结合患者临床病理特征分析RKIP的表达水平与Ⅱ期结直肠癌患者预后的关系。结果:结直肠癌组织中RKIP的表达水平显著低于相应癌旁组织(P<0.001)。与无复发转移患者相比,复发转移患者癌组织中RKIP的表达明显降低(P=0.034)。RKIP高表达组的10年无疾病进展生存率明显优于RKIP低/中表达组(89% vs 75%,P=0.032);与RKIP低/中表达组相比,RKIP高表达组的10年总生存期有延长趋势,但无统计学意义(71%vs88%,P=0.058)。RKIP表达下降(HR 0.37;95%CI:0.13～0.99;P=0.040)和术前CEA水平升高(HR 3.50;95%CI:1.30～9.37;P=0.013)是Ⅱ期结直肠癌患者术后复发转移的危险因素。结论:RKIP在Ⅱ期结直肠癌原发肿瘤组织中表达下调,高表达患者无疾病进展生存期明显延长,提示RKIP表达水平是Ⅱ期结直肠癌患者术后复发转移的重要预后因素。展开更多
基金Supported by a grant from the Qingdao Public Sphere Sci-technical Support Project(No.09-1-1-13-nsh)
文摘Objective： The purpose of the study is to investigate the effects of up-regulation of Raf kinase inhibitor protein （RKIP） on the chemosensitivity of cervical cancer Hela cells. Methods： Eukaryotic expression plasmid pcDNA3.1（±）-ssRKIP containing human overall length RKIPcDNA was transfected into cervical cancer Hela cell by lipofectin assay, establishing a stable cell line containing a target gene by G418. Expression of RKIP in Hela cells was measured by Western blot analysis. After treatment with cisplatin of different concentrations and intervals of time, the effect of RKIP on the proliferation of Hela cells was evaluated by MTT method. The flow cytometry was used to investigate whether the RKIP could inhibit apoptosis in Hela cells induced by cisplatin. Results： The expression of RKIP in Hela cells transfected with pcDNA3.1-ssRKIP was increased obviously. After different concentrations of cisplatin treatment cells for 24, 48 and 72 h, the growth inhibition rate in Hela cells transfected with pcDNA3.1-ssRKIP was significantly higher than in control cells （P 〈 0.05）. With 5 pg/mL cisplatin treatment for 24 h, pcDNA3.1-ssRKIP-transfected Hela cells had an obviously higher percentage of apoptosis （23.2 ± 0.24）% than non-transfected cells （12.4 ± 0.31）% and empty vector-transfected cells （13.4 ± 0.47）%. Without treatment of cisplatin, the percentage of apoptosis for Hela cells transfected with pcDNA3.1-ssRKIP was （5.7 ± 0.12）%, which was still higher than those of the non-transfected cells （2.9 ± 0.21）% and empty vector-transfected cells （3 ± 0.08）%. Conclusion： Higher expres- sion of RKIP gene can improve chemosensitivitv of cervical cancer Hela cells to cisplatin.
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。
文摘目的:探讨RKIP在Ⅱ期结直肠癌中的表达及其与患者预后的关系。方法:收集130例Ⅱ期结直肠癌患者的术后组织标本,采用免疫组织化学SP法检测RKIP在原发肿瘤组织及癌旁组织中的表达,结合患者临床病理特征分析RKIP的表达水平与Ⅱ期结直肠癌患者预后的关系。结果:结直肠癌组织中RKIP的表达水平显著低于相应癌旁组织(P<0.001)。与无复发转移患者相比,复发转移患者癌组织中RKIP的表达明显降低(P=0.034)。RKIP高表达组的10年无疾病进展生存率明显优于RKIP低/中表达组(89% vs 75%,P=0.032);与RKIP低/中表达组相比,RKIP高表达组的10年总生存期有延长趋势,但无统计学意义(71%vs88%,P=0.058)。RKIP表达下降(HR 0.37;95%CI:0.13～0.99;P=0.040)和术前CEA水平升高(HR 3.50;95%CI:1.30～9.37;P=0.013)是Ⅱ期结直肠癌患者术后复发转移的危险因素。结论:RKIP在Ⅱ期结直肠癌原发肿瘤组织中表达下调,高表达患者无疾病进展生存期明显延长,提示RKIP表达水平是Ⅱ期结直肠癌患者术后复发转移的重要预后因素。
