siRNA阻断PRKCι表达对人卵巢癌细胞株SKOV3迁移和侵袭的影响

发布时间：2018-06-07 14:47

本文选题：PRKCι + 卵巢癌　；参考：《现代妇产科进展》2015年09期

【摘要】：目的:探讨PRKCι在卵巢癌迁移和侵袭中的作用。方法:生物信息学分析PRKCι在卵巢癌中的表达,Real-time PCR和Western blot法检测卵巢癌细胞株中PRKCι的表达情况,设计针对PRKCι的siRNA并转染SKOV3卵巢癌细胞,观察转染后SKOV3细胞中PRKCι基因表达情况,以及SKOV3细胞的迁移、侵袭能力的变化,并检测PRKCι基因敲减后其下游转移相关效应分子MMP10的表达。结果:PRKCι在卵巢癌芯片(TCGA和GDS3592)中的表达与正常卵巢组织比较,表达差异2倍(P0.0001和P=0.0078)。与HOSE细胞相比,ES2、CAOV3、OVCAR3、SKOV3、A2780卵巢癌细胞株中存在PRKCιmRNA和蛋白水平的高表达,其中SKOV3细胞中PRKCι表达水平最高。PRKCι特异性的siRNA-1和siRNA-2均能有效抑制SKOV3细胞中PRKCιmRNA和蛋白表达,mRNA水平抑制效率分别为(80.5±10.23)%、(74.6±12.48)%(P0.01),在蛋白水平抑制效率分别为(71.37±11.34)%、(68.22±12.19)%(P0.05)。细胞划痕实验显示,PRKCιsiRNA明显降低了SKOV3细胞的迁移能力,抑制效率分别为(54.31±12.87)%、(45.25±14.02)%(P0.05);Transwell实验显示,PRKCιsiRNA明显降低了SKOV3细胞的侵袭能力,分别降低了57.48%和38.38%(P0.05);PRKCιsiRNA明显抑制了转移相关效应分子MMP10在mRNA和蛋白水平表达,mRNA水平分别下调(56.76±13.83)%、(52.99±14.38)%(P0.05),蛋白水平分别下调(35.65±9.10)%、(37.14±14.26)%(P0.05)。结论:PRKCι在卵巢癌中高表达,是参与卵巢癌转移的重要基因,可能作为抑制卵巢癌转移的新的靶向分子。
[Abstract]:Objective: to investigate the role of PRKC l in the migration and invasion of ovarian cancer. Methods: bioinformatics was used to detect the expression of PRKC l in ovarian cancer cell lines by real-time PCR and Western blot. The siRNA targeting PRKC l was designed and transfected into SKOV3 ovarian cancer cells. The expression of PRKC l gene was observed in SKOV3 cells after transfection. And the migration and invasion ability of SKOV3 cells, and the expression of the downstream metastasis related effector molecule (MMP10) of PRKC l gene knockout was detected. Results compared with normal ovarian tissues, the expression of 1% PRKC l in ovarian cancer microarray TCGA and GDS3592 was 2 times higher than that in normal ovarian tissues (P0.0001 and P0. 0078). Compared with HOSE cells, there was a high expression of PRKC mRNA and protein in OVCAR3SKOV3A2780 ovarian cancer cell line. Both siRNA-1 and siRNA-2, which had the highest expression level of PRKC siRNA-1 and siRNA-2, could effectively inhibit the expression of PRKC mRNA and protein in SKOV3 cells. The inhibitory efficiency of siRNA-1 and siRNA-2 were 74.6 卤12.48% and 71.37 卤11.34 卤68.22 卤12.19, respectively, and that of P0.05 in SKOV3 cells were 74.6 卤12.48 and 71.37 卤11.34, 68.22 卤12.19, respectively. The cell scratch assay showed that PRKC l siRNA significantly decreased the migration ability of SKOV3 cells, and the inhibitory efficiency was 54.31 卤12.87%, respectively. The inhibition efficiency was 45.25 卤14.02%. The results showed that PRKC l siRNA significantly decreased the invasion ability of SKOV3 cells. 57.48% and 38.38% respectively decreased the expression of MMP10 in mRNA and protein level by 57.48%, 38.38% and 38.38%, respectively, and down-regulated the expression of MMP10 at the level of mRNA and protein, respectively, 56.76 卤13.83% and 52.99 卤14.38, respectively, and the protein level was down regulated by 35.65 卤9.10 and 37.14 卤14.26, respectively. Conclusion the high expression of the protein PRKC in ovarian cancer is an important gene involved in ovarian cancer metastasis, which may be a new target molecule for inhibiting ovarian cancer metastasis.
【作者单位】：苏州大学附属第一医院妇产科;
【基金】：国家自然科学青年基金(No:81302275) 苏州市科技局项目(No:SYS201463)
【分类号】：R737.31

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