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稳定转染PGRMC1基因的小分子干扰RNA对卵巢癌细胞体外增殖的影响[J]. 肿瘤防治研究, 2013, 40(07): 652-655. DOI: 10.3971/j.issn.1000-8578.2013.07.006
引用本文: 稳定转染PGRMC1基因的小分子干扰RNA对卵巢癌细胞体外增殖的影响[J]. 肿瘤防治研究, 2013, 40(07): 652-655. DOI: 10.3971/j.issn.1000-8578.2013.07.006
Effects of Small Interfering RNA Specific to Stable Transfection Gene of PGRMC1 on Proliferation of Ovarian Carcinoma in vitro[J]. Cancer Research on Prevention and Treatment, 2013, 40(07): 652-655. DOI: 10.3971/j.issn.1000-8578.2013.07.006
Citation: Effects of Small Interfering RNA Specific to Stable Transfection Gene of PGRMC1 on Proliferation of Ovarian Carcinoma in vitro[J]. Cancer Research on Prevention and Treatment, 2013, 40(07): 652-655. DOI: 10.3971/j.issn.1000-8578.2013.07.006

稳定转染PGRMC1基因的小分子干扰RNA对卵巢癌细胞体外增殖的影响

Effects of Small Interfering RNA Specific to Stable Transfection Gene of PGRMC1 on Proliferation of Ovarian Carcinoma in vitro

  • 摘要: 目的 观察稳定转染PGRMC1基因的小分子干扰RNA(siRNA)对卵巢癌COC1细胞体外增殖的影响。 方法 实验分为3组,重组载体组即由pcDNA3.1A- PGRMC1质粒转染COC1细胞;空载体组即仅转染空载体pcDNA3.1A质粒;空白组即未转染质粒组。3组均经抗性筛选获得稳定转染的细胞系。采用RT-PCR技术检测3组细胞中PGRMC1 mRNA的表达;四甲基偶氮唑盐比色法观察细胞增殖情况,绘制细胞生长曲线;裸鼠接种3组细胞后观察其成瘤能力的变化。 结果 (1)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)%,两者比较,差异有统计学意义(P<0.05);空载体组和空白组PGRMC1 mRNA转染前、后表达水平分别比较,差异无统计学意义(P>0.05)。(2)重组载体组COC1细胞增殖速度明显慢于空载体组和空白对照组(P<0.05);(3)接种转染后的COC1细胞后,重组载体组裸鼠的成瘤时间为(5.8±0.7)d,明显短于空载体组的(11.9±0.5)d和空白对照组的(12.6±0.9)d,差异均有统计学意义(P<0.05);而空载体组与空白对照组比较,差异无统计学意义(P>0.05)。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±0.4)g和(197±26) mm3,明显低于空载体组(2.3±0.4)g和(785±38)mm3 和空白对照组为(2.5±0.8) g和(896±22) mm3 ,差异均有统计学意义(P<0.05)。 结论 PGRMC1 siRNA可显著降低卵巢癌细胞中的PGRMC1 表达,并对卵巢癌细胞的生长有抑制作用。

     

    Abstract: Objective To investigate the effects of the proliferation of the human ovarian cell line (COC1) by stable transfection short hairpin RNA into the target PGRMC1 gene. Methods The text subjects were divided into three groups. PGRMC1 recombinant expression plasmid was constructed and then was transfected into COC1 cells by lipofectamine 2000. The mock-vehicle group only transfected the blank load pcDNA3.1 A plasmid. And the blank group was composed of non-transfected plasmids. The three groups gained stable-transfected cell lines after resistance selection. The expression of PGRMC1 mRNA was tested by reverse transcription RT-PCR.The proliferation of COC1 cells after PGRMC1 gene transfection was measured by methylthiazolyl tetrazolium(MTT). And cells growth curves were plotted. The tumor growth of the nude mice inoculated of tumor cells was compared with before and after transfection. Results The results of RT-PCR showed that the significant expressions of PGRMC1 mRNA after infected by PGRMC1 plasmid were higher than those in the control. The COC1 cell growth in stable transfection status was more significantly decreased than that in non-transfection status (P<0.05). After inoculating transfected cells into nude mice,it took (5.8±0.7 )days to grow the planed tumors in group infected by PGRMC1 plasmid. The days were significantly shorter than those in the control(11.9±0.5)days and in the blank group (12.6±0.9) days (P<0.05). There were no significant differences between the mock-vehicle group and the blank group (P>0.05). And five weeks after inoculation, mice tumors' weights and sizes of the recombinant vectors group were (0.7±0.4) g and (197±26) mm3 respectively, which were significantly lower than those of the mock-vehicle group (2.3±0.4) g and (785±38)mm3 and those of the blank control group(2.5±0.8) g and (896±22)mm3. Conclusion PGRMC1 siRNA could remarkably reduce the expression of PGRMC1 gene in ovarian cell and also inhabit the ovarian cell growth.

     

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