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[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±26mm~3,明显低于空载体组(2.3±0.4g和785±38mm~3)和空白对照组的(2.5±0.8g和896±22mm~3]。[结论 ]PGRMC1 si RNA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。
[Objective] To investigate the effect of PGRMC1 gene small interfering RNA on the proliferation of ovarian cancer cells in vitro. [Method] The expression of PGRMC1 protein and mRNA in paraffin sections of different ovarian lesions (ovarian cancer, benign ovarian tumor, borderline ovarian tumor and normal ovary) were detected by immunohistochemistry and real-time quantitative PCR. Cancer model, PGRMC1 gene was specifically knocked out by using RNA interference technology and the change of PGRMC1 gene activity was detected. [Results] (1) The results of immunohistochemistry showed that the expression of PGRMC1 protein in borderline ovarian tumor and ovarian cancer group was significantly higher than that in normal ovarian group and benign ovarian tumor group (P <0.01); (2) PCR results showed that the expression of PGRMC1 mRNA in ovarian cancer group was significantly higher than that in benign ovarian disease group (0.936 ± 0.725) (3.526 ± 1.386), PGRMC1 mRNA expression in ovarian cancer group was 3.51 times that of benign ovarian tumor group P <0.01). (3) The expression levels of PGRMC1 mRNA in the recombinant vector group before and after transfection were 79.6% ± 2.3% and 29.4% ± 1.6%, respectively, with statistical significance (P <0.05). The empty vector group and blank control group PGRMC1 mRNA expression before and after transfection was no significant difference (P> 0.05). (4) After 5 weeks of inoculation, the tumor weight and volume of the nude mice in the recombinant vector group were 0.7 ± 0.4g and 197 ± 26mm 3, respectively, significantly lower than that in the empty vector group (2.3 ± 0.4g and 785 ± 38mm 3) and (2.5 ± 0.8g and 896 ± 22mm ~ 3) in blank control group. [Conclusion] PGRMC1 si RNA can significantly reduce the expression of PGRMC1 in ovarian cancer cells and inhibit the proliferation of ovarian cancer cells in vitro.