现代妇产科进展Issue(9)：651-655,5.DOI:10.13283/j.cnki.xdfckjz.2015.09.003
siRNA阻断PRKCɩ表达对人卵巢癌细胞株SKOⅤ3迁移和侵袭的影响
PRKCɩ knock down by siRNA inhibits migration and invasion in ovarian cancer cell line ;SKOV3
摘要
Abstract
Objective:To investigate the role of PRKCɩ in migration and invasion of o-varian cancer;Methods:PRKCɩ 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 PRKCɩ 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. Re-sults:Analysis of microarray data showed that PRKCɩ 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 PRKCɩ 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 PRKCɩ in these cell lines. PRKCɩ targeting siRNA-1 and siRNA-2 both effectively knocked down PRKCɩ 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 mi-gration 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, PRKCɩ 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:PRKCɩ is proba-bly involved in ovarian cancer migration and invasion, and serves as a candidate of targeted therapeutics inhibiting ovarian cancer metastasis.关键词
PRKC咨/卵巢癌/迁移/侵袭/MMP10Key words
PRKC iota( PRKCɩ)/Ovarian cancer/Migration/Ⅰnvasion/MMP10分类
医药卫生引用本文复制引用
蒯玲玲,周金华,沈芳荣,李珉,陈友国..siRNA阻断PRKCɩ表达对人卵巢癌细胞株SKOⅤ3迁移和侵袭的影响[J].现代妇产科进展,2015,(9):651-655,5.基金项目
国家自然科学青年基金(No:81302275)及苏州市科技局项目(No:SYS201463) （No:81302275）
