siRNA阻断PRKCι表达对人卵巢癌细胞株SKOV3迁移和侵袭的影响  

PRKCι knock down by siRNA inhibits migration and invasion in ovarian cancer cell line SKOV3

作　　者：蒯玲玲 周金华[1] 沈芳荣[1] 李珉[1] 陈友国[1] 

机构地区：[1]苏州大学附属第一医院妇产科,苏州215006

出　　处：《现代妇产科进展》2015年第9期651-655,共5页Progress in Obstetrics and Gynecology

基　　金：国家自然科学青年基金(No:81302275);苏州市科技局项目(No:SYS201463)

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(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 PRKCι in migration and invasion of ovarian cancer; Methods： PRKCι expression was bioinformatically analyzed in open accessed gene expression profiles of ovarian cancer and validated in ovarian cancer cell lines by real-time PCR as well as Western blot,after PRKCι knocking down by targeting siRNAs,the migration and invasion ability was assessed by wound healing and transwell assay in SKOV3 cells. The downstream effector gene MMP10 was also determined by real-time PCR and Western blot. Results： Analysis of microarray data showed that PRKCι mRNA was increased by more than 2-fold in ovarian cancer compared with normal ovary（ P〈0. 0001 in TCGA data,P = 0. 0078 in GDS3592 data）. Besides,elevated PRKCι mRNA and protein expression was also demonstrated in ES2,CAOV3,OVCAR3,SKOV3 and A2780 ovarian cancer cell lines. SKOV3 showed the most robust expression of PRKCι in these cell lines. PRKCι targeting siRNA-1 and siRNA-2both effectively knocked down PRKCι expression. The respective inhibitory efficacy were（ 80. 5±10. 23） %,（ 74. 6 ±12. 48） %（ P〈0. 01） at mRNA level and（ 71. 37 ±11. 34） %,（ 68. 22 ±12. 19） %（ P〈0. 05） at protein level. Wound healing assay showed that siRNA attenuated migration potential of SKOV3 cells by（ 54. 31 ±12. 87） % for siRNA-1 and（ 45. 25 ±14. 02） %for siRNA-2（ P〈0. 05）. Furthermore,siRNA-1 and siRNA-2 blocked the invasion potential of SKOV3 cells by 57. 48% and 38. 38% respectively（ P〈0. 05） in transwell assay. In addition,PRKCι knockdown led to the decreased expression of MMP10 at mRNA by（ 56. 76 ±13. 83） %（ P〈0. 05） and protein level by（ 52. 99±14. 38） %（ P〈0. 05）. Conclusions： PRKCι is probably involved in ovarian cancer migration and invasion,and serves as a candidate of targeted therapeutics inhibiting ovarian cancer metastasis.

关 键 词：PRKCι 卵巢癌 迁移 侵袭 MMP10 

分 类 号：R737.33[医药卫生—肿瘤]