宋晓霞,姚 丽,张占薪.PGRMC1基因小分子干扰RNA对卵巢癌细胞体外增殖的影响[J].肿瘤学杂志,2017,23(6):508-512.
PGRMC1基因小分子干扰RNA对卵巢癌细胞体外增殖的影响
Effect of Small Interfering RNA Silencing PGRMC1 Gene on Proliferation of Ovarian Carcinoma in vitro and in vivo
投稿时间:2016-10-29  
DOI:10.11735/j.issn.1671-170X.2017.06.B010
中文关键词:  卵巢癌  PGRMC1  siRNA  转染
英文关键词:ovarian cancer  progesterone receptor membrane component-1  small interfering RNA  transfection
基金项目:
作者单位
宋晓霞 郑州人民医院 
姚 丽 郑州人民医院 
张占薪 郑州人民医院 
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中文摘要:
      摘 要:[目的] 探讨PGRMC1基因小分子干扰RNA对卵巢癌细胞体外增殖的影响。[方法] 首先利用免疫组化和实时定量PCR方法检测不同卵巢病变组织(卵巢癌、良性卵巢肿瘤、交界性卵巢肿瘤和正常卵巢)蜡块中PGRMC1蛋白及mRNA表达水平;进一步通过构建裸鼠卵巢癌模型,利用RNA干扰技术将PGRMC1基因特异性敲除,检测由此导致的PGRMC1基因活性的改变。[结果] (1)免疫组化结果显示,与正常卵巢组及良性卵巢肿瘤组相比,卵巢交界性肿瘤组和卵巢癌组PGRMC1蛋白的表达显著增高(P<0.01);(2)实时定量PCR方法检测结果显示,与良性卵巢病变组(0.936±0.725)相比,卵巢癌组中(3.526±1.386)PGRMC1 mRNA的表达显著增高,卵巢癌组PGRMC1 mRNA表达是良性卵巢肿瘤组的3.51倍(P<0.01)。(3)重组载体组细胞中PGRMC1 mRNA转染前、后的表达水平分别为79.6%±2.3%和29.4%±1.6%,差异有统计学意义(P<0.05),空载体组和空白对照组PGRMC1 mRNA转染前、后表达水平差异无统计学意义(P>0.05)。(4) 接种5周后,重组载体组裸鼠的肿瘤重量和体积分别为0.7±0.4g和197±26mm3,明显低于空载体组(2.3±0.4g和785±38mm3)和空白对照组的(2.5±0.8g和896±22mm3]。[结论] PGRMC1 siRNA可显著降低卵巢癌细胞中的PGRMC1的表达,并对卵巢癌细胞的体外增殖有抑制作用。
英文摘要:
      Abstract:[Objective] To investigate the effect of small interfering RNA silencing PGRMC1 gene on proliferation of ovarian carcinoma in vitro and in vivo. [Methods] The expression of PGRMC1 protein and mRNA were detected by the immunohistochemical technique and real-time quantitative PCR in ovarian cancer,benign ovarian lesions and normal ovarian tissues. Human ovarian carcinoma COC1 cells were inoculated in nude mice,and the PGRMC1 gene was silenced by RNA interference in vivo. The changes of PGRMC1 expression were detected. [Results] Immunohistochemical results showed that compared with normal ovarian and benign ovarian tumors,PGRMC1 protein expression in borderline ovarian tumors and ovarian cancer was significantly higher(P<0.01). Real-time quantitative PCR results showed that compared with benign ovarian lesions(0.936±0.725),PGRMC1 mRNA expression in ovarian cancer(3.526±1.386) was significantly higher (P<0.01). The expression level of PGRMC1 mRNA in COC1 cells was reduced from 79.6%±2.3% to 29.4%±1.6% after transfected with siRNA targeting to PGRMC1(P<0.05). Five weeks after inoculation,the tumor weight and volume in nude mice with silenced HPGRMC1 gene were 0.7±0.4g and 197±26mm3,which were significantly lower than those in the empty vector group (2.3±0.4g and 785±38mm3) and blank group (2.5+0.8g and 896±22mm3) (P<0.05).[Conclusion] HPGRMC1 siRNA can significantly reduce the expression of PGRMC1 in ovarian cancer cells and inhibit the proliferation of ovarian cancer cells in vitro and in vivo.
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