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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 the invasion, proliferation and cloning ability of ovarian cancer cell line SKOV3 and provide a new theoretical basis for targeted therapy of ovarian cancer. Methods: SKOV3 cells were transfected with shCD26, shGAPDH and shNC (empty plasmid) respectively. The transfection efficiency of three groups of cells was calculated by intracellular cytoplasm green fluorescence signal. The expression of DPPⅣmRNA and protein in cells before and after transfection were detected by RT-PCR and Western blot respectively. The invasion ability of cells was detected by invasion assay, the cell cycle was detected by flow cytometry, the proliferation and cloning ability of cells were detected by CCK-8 and plate clone formation assay . Results: The transfection efficiency of shCD26, shGAPDH and shNC cells were 59%, 64% and 60% respectively, with no significant difference (P> 0.05). The relative expression levels of DPPⅣ mRNA in shNC, shCD26, shGAPDH and non-tranfected groups were 1.20 ± 0.11, 0.80 ± 0.01, 2.15 ± 0.15 and 1.32 ± 0.04, respectively. The expression of DPPⅣmRNA and protein in shCD26 group were significantly lower than The remaining three groups (P <0.05). The numbers of transmembrane cells in shCD26 group, shNC group and non-transfected group were 37 ± 4.08, 65 ± 4.74 and 66.8 ± 3.82, respectively, and the numbers of clones were 8.3 ± 0.57, 13.33 ± 1.52 and 14.0 ± 1.0sh, respectively. The numbers of transmembrane cells, number of cloned cells and cell viability in shCD26 group were significantly lower than those in shNC group and non-transfected group (P <0.05). Compared with shNC group and nontransfected group, the ratio of G0 / G1 phase in shCD26 group increased, while the ratio of S and G2 / M phase decreased. The difference was significant (P <0.05). Conclusion: DPP Ⅳ may promote SKOV3 cell invasion, proliferation and colony formation.