沉默二肽基肽酶Ⅳ/CD26对上皮性卵巢癌SKOV3细胞侵袭及增殖的影响  

The effect of silencing Dipeptidyl peptidase Ⅳ on invasiveness and proliferation of SKOV3 epithelial ovarian cancer cells

作　　者：马艳美[1] 陈志华[1] 谢娅[1] 苏玥辉[1] 张毅 张梦真[1] 

机构地区：[1]郑州大学第一附属医院妇产科,郑州450052 [2]河南省临床医学重点实验室,郑州450052

出　　处：《现代妇产科进展》2014年第6期440-444,共5页Progress in Obstetrics and Gynecology

基　　金：河南省杰出青年基金资助项目(No:104100510007);河南省医学科技攻关重点项目(No:201001005)

摘　　要：目的:探讨沉默二肽基肽酶Ⅳ(DPPⅣ/CD26)对卵巢癌SKOV3细胞侵袭、增殖及克隆能力的影响,为卵巢癌靶向治疗提供新的理论依据。方法:采用shCD26、shGAPDH及shNC(空质粒)分别转染SKOV3细胞,通过细胞质内绿色荧光信号计算3组细胞的转染效率。RT-PCR及Western blot分别检测转染前后细胞中DPPⅣmRNA及蛋白的表达,基底膜侵袭实验检测细胞侵袭能力,流式细胞检测细胞周期变化,CCK-8及平板克隆形成实验检测细胞增殖及克隆能力。结果:shCD26、shGAPDH及shNC组细胞的转染效率分别为59%、64%和60%;差异无统计学意义(P>0.05)。shNC、shCD26、shGAPDH及未转染组(non-tranfected)组中DPPⅣmRNA相对表达量分别为1.20±0.11、0.80±0.01、2.15±0.15和1.32±0.04;shCD26组中DPPⅣmRNA和蛋白表达均显著低于其余3组(P<0.05)。shCD26组、shNC组和non-transfected组的穿膜细胞数分别为37±4.08、65±4.74和66.8±3.82,克隆数分别为8.3±0.57、13.33±1.52和14.0±1.0sh。shCD26组的穿膜细胞数、克隆数及细胞活力均显著低于shNC组、non-transfected组(P<0.05)。shCD26组细胞的G0/G1期比率增加,而S及G2/M期比率降低,与shNC组、nontransfected组比较,差异显著(P<0.05)。结论:DPPⅣ可能具有促进SKOV3细胞侵袭、增殖及克隆形成等作用。Objective：To investigate the effect of silencing dipeptidyl peptidase IV （DPPIV/CD26）on invasiveness, proliferation and colony formation in SKOV3 ovarian cancer cells, and therefore provide evidence for targeted therapy of ovarian cancer. Methods. SKOV3 was transfected with short hairpin RNA （shRNA） of DPP IV, GAPDH and empty plasmid （ shCD26, shGAPDH, shNC）, respectively. Transfection efficiency of the short hairpin RNA was estimated according to the amount of green fluorescence by fluorescence microscope. The ex- pression of DPPIV mRNA and protein were examined by reverse transcription PCR and western blot, respectively. The Invasiveness, cell cycle, cell proliferation and the colony fo^tnation of SK- OV3 cells were analysed using Matrigel invasion assay,/low cytometry, Cell Counting Kit-8 （CCK-8） and colony formation assay, respectively. Results： The transfection efficiency of shCD26,shGAPHD and shNC were 59% ,64% and 60% （P〉0.05） , and the difference were not of with statistical significance. The expression levels of DPP IV mRNA of shNC, shCD26, shGAPDH and non-transfeeted were 0. 80±0. 01,1. 20±0.11,2.15 ±0.15 and 1.32±0.04. The level of DPPIV protein of the shCD26-transfeeted was signifieantly lower than the other three groups（P〈0.05 ）. The cell numbers of transmembrane of shCD26, shNC and non-transfeeted were respectively 37 ±4.08,65 ±4.74 and 66.8±3.82, and the number of the colony formation were 8.3±0.57,13.33 ± 1.52 and 14.0±1.0. The cell number of transmembrane, the rate of the colony formation and the cell activity of the shCD26 were considerably below the shNC and the non-transfected （P〈0.05）. The shCD26-transfeeted cells demonstrated higher Go/G1 propertion, but contained S-phase cells and Gz/M-phase cells at lower levels. Compared to the shNC and the non-transfeeted, the difference was significant （P〈0.05）. Conclusion： DPPIV may be involved in cell invasiveness;Proliferation and colony formation of SKOV3 eells.

关 键 词：卵巢肿瘤 SKOV3 DPPⅣ CD26 短链发夹RNA 

分 类 号：R737.33[医药卫生—肿瘤]