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目的探讨血管内皮生长因子受体2(kinase insert domain-containing receptor,KDR)反义寡核苷酸(antisense oligode-oxynucleotides,ASODN)对人前列腺癌PC-3细胞的增殖调控作用。方法设计并合成KDR的正义、反义寡核苷酸,经脂质体介导转染人前列腺癌PC-3细胞。用噻唑盐法观察不同浓度(0.01,0.05,0.1,0.2,0.4μmol/L)正义、反义寡核苷酸及阳离子脂质体对肿瘤细胞增殖的抑制作用,RT-PCR方法检测不同浓度(0.01,0.05,0.1,0.2,0.4μmol/L)ASODN转染后KDRmRNA的表达情况,流式细胞仪分析细胞周期分布和细胞凋亡。结果转染ASODN后48 h达抑制高峰,低浓度(0.01μmol/L)即发生抑制,抑制强度与反义寡核苷酸浓度有相关性,而正义寡核苷酸和阳离子脂质体对PC-3细胞的增殖无显著影响。转染KDR反义寡核苷酸各浓度组细胞中KDR mRNA的表达都不同程度地降低,各浓度组与空白对照组相比KDR mRNA表达的差异有统计学意义。各组均出现不同程度的细胞凋亡,但各浓度组间细胞周期分布无显著性差异。结论 KDR基因在促进人前列腺癌PC-3细胞增殖中发挥着一定的作用,有望成为治疗雄激素非依赖性前列腺癌的分子靶点。
Objective To investigate the effect of antisense oligodeoxynucleotides (ASODN) on the proliferation of human prostate cancer PC-3 cells. Methods The sense and antisense oligonucleotides of KDR were designed and synthesized and transfected into human prostate cancer PC-3 cells by liposome. The inhibitory effects of different concentrations (0.01,0.05,0.1,0.2,0.4μmol / L) of antisense, antisense oligonucleotide and cationic liposome on the proliferation of tumor cells were observed by thiazolium salt method. The effects of different concentrations ( 0.01, 0.05, 0.1, 0.2, 0.4μmol / L) ASODN transfected KDRmRNA expression, cell cycle distribution and apoptosis analysis by flow cytometry. RESULTS: The inhibition peak was reached at 48 h after ASODN transfection and was inhibited at low concentration (0.01 μmol / L). The inhibitory strength was correlated with the concentration of antisense oligonucleotide, while the sensitivity of antisense oligodeoxynucleotides and cationic liposomes to PC -3 cells proliferation no significant effect. The expression of KDR mRNA in KDR antisense oligonucleotide transfected cells decreased to different extents, and the difference of KDR mRNA expression between each concentration group and blank control group was statistically significant. There were different degrees of apoptosis in each group, but there was no significant difference in the cell cycle distribution between the groups. Conclusion KDR gene plays a role in promoting the proliferation of human prostate cancer PC-3 cells and is expected to become a molecular target for the treatment of androgen-independent prostate cancer.