论文部分内容阅读
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(P<0.0001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)%(P<0.01),在蛋白水平抑制效率分别为(71.37±11.34)%、(68.22±12.19)%(P<0.05)。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)%(P<0.05);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%(P<0.05);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)%(P<0.05),蛋白水平分别下调(35.65±9.10)%、(37.14±14.26)%(P<0.05)。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。
Objective: To investigate the role of PRKC1 in the migration and invasion of ovarian cancer. Methods: The expression of PRKC1 in ovarian cancer was analyzed by bioinformatics method. The expression of PRKC1 in ovarian cancer cell lines was detected by Real-time PCR and Western blot. The siRNA targeting PRKC1 was transfected into SKOV3 ovarian cancer cells. After transfection, The expression of PRKC1 gene in SKOV3 cells and the migration and invasion ability of SKOV3 cells were detected. The expression of MMP10, a downstream effector of PRKC1 gene knockdown, was detected. Results: The expression of PRKC1 in ovarian cancer cells (TCGA and GDS3592) was> 2 times higher than that in normal ovarian tissue (P <0.0001 and P = 0.0078). Compared with HOSE cells, the expression of PRKC1 mRNA and protein in ES2, CAOV3, OVCAR3, SKOV3 and A2780 ovarian cancer cell lines was high, and the expression level of PRKC1 in SKOV3 cells was the highest. Both PRKC1-specific siRNA-1 and siRNA-2 were able to effectively inhibit the expression of PRKC1 mRNA and protein in SKOV3 cells, and the inhibitory rates of mRNA levels were (80.5 ± 10.23)% and (74.6 ± 12.48)% The inhibition efficiencies were (71.37 ± 11.34)% and (68.22 ± 12.19)%, respectively (P <0.05). Cell scratch assay showed that PRKC1siRNA significantly reduced the migration ability of SKOV3 cells, and the inhibitory efficiency was (54.31 ± 12.87)% and (45.25 ± 14.02)% respectively (P <0.05). Transwell assay showed that PRKC1siRNA significantly decreased the SKOV3 cell migration (P <0.05). PRKC1siRNA significantly inhibited the expression of MMP10 at mRNA and protein levels (56.76 ± 13.83%, (52.99 ± 14.38)%, respectively) (P <0.05). The protein levels were decreased by 35.65 ± 9.10% and 37.14 ± 14.26%, respectively (P <0.05). Conclusion: PRKC1 is overexpressed in ovarian cancer and is an important gene involved in ovarian cancer metastasis. It may serve as a new target molecule that inhibits the metastasis of ovarian cancer.