siRNA阻斷PRKCι表達對人卵巢癌細胞株SKOV3遷移和侵襲的影響
本文選題:PRKCι + 卵巢癌; 參考:《現(xiàn)代婦產科進展》2015年09期
【摘要】:目的:探討PRKCι在卵巢癌遷移和侵襲中的作用。方法:生物信息學分析PRKCι在卵巢癌中的表達,Real-time PCR和Western blot法檢測卵巢癌細胞株中PRKCι的表達情況,設計針對PRKCι的siRNA并轉染SKOV3卵巢癌細胞,觀察轉染后SKOV3細胞中PRKCι基因表達情況,以及SKOV3細胞的遷移、侵襲能力的變化,并檢測PRKCι基因敲減后其下游轉移相關效應分子MMP10的表達。結果:PRKCι在卵巢癌芯片(TCGA和GDS3592)中的表達與正常卵巢組織比較,表達差異2倍(P0.0001和P=0.0078)。與HOSE細胞相比,ES2、CAOV3、OVCAR3、SKOV3、A2780卵巢癌細胞株中存在PRKCιmRNA和蛋白水平的高表達,其中SKOV3細胞中PRKCι表達水平最高。PRKCι特異性的siRNA-1和siRNA-2均能有效抑制SKOV3細胞中PRKCιmRNA和蛋白表達,mRNA水平抑制效率分別為(80.5±10.23)%、(74.6±12.48)%(P0.01),在蛋白水平抑制效率分別為(71.37±11.34)%、(68.22±12.19)%(P0.05)。細胞劃痕實驗顯示,PRKCιsiRNA明顯降低了SKOV3細胞的遷移能力,抑制效率分別為(54.31±12.87)%、(45.25±14.02)%(P0.05);Transwell實驗顯示,PRKCιsiRNA明顯降低了SKOV3細胞的侵襲能力,分別降低了57.48%和38.38%(P0.05);PRKCιsiRNA明顯抑制了轉移相關效應分子MMP10在mRNA和蛋白水平表達,mRNA水平分別下調(56.76±13.83)%、(52.99±14.38)%(P0.05),蛋白水平分別下調(35.65±9.10)%、(37.14±14.26)%(P0.05)。結論:PRKCι在卵巢癌中高表達,是參與卵巢癌轉移的重要基因,可能作為抑制卵巢癌轉移的新的靶向分子。
[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 鹵12.48% and 71.37 鹵11.34 鹵68.22 鹵12.19, respectively, and that of P0.05 in SKOV3 cells were 74.6 鹵12.48 and 71.37 鹵11.34, 68.22 鹵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 鹵12.87%, respectively. The inhibition efficiency was 45.25 鹵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 鹵13.83% and 52.99 鹵14.38, respectively, and the protein level was down regulated by 35.65 鹵9.10 and 37.14 鹵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.
【作者單位】: 蘇州大學附屬第一醫(yī)院婦產科;
【基金】:國家自然科學青年基金(No:81302275) 蘇州市科技局項目(No:SYS201463)
【分類號】:R737.31
【共引文獻】
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