白介素-28 A增加顺铂对卵巢癌细胞毒性的作用及机制研究

目的:探讨白介素-28A(IL-28A)增强顺铂对卵巢癌的细胞毒性作用及其机制.方法:实时荧光定量PCR法检测卵巢癌组织和细胞及正常卵巢上皮组织和细胞中IL-28A水平.IL-28A(100ng/ml)预处理C13K细胞24h,CCK-8和流式细胞术分别检测顺铂对C13K细胞半数抑制浓度(IC50)、细胞存活率和细胞凋...

IL-28A enhances the cytotoxicity of cisplatin against ovarian carcinoma cells and its possible mechanism

Objective:To investigate whether IL-28A can enhance the cytotoxicity of cisplatin against ovarian carcinoma cells C13K and the possible mechanisms.Methods:Q-PCR was applied to detect the expression of IL-28A in the ovarian carcinoma tissues or cells,ovary normal epithelium tissues or cells,respectively.Pretreated with IL-28A(100ng/ml)for 24h,then treat C13K cells with cisplatin for 24h.Cell viability and the half concentration(IC50)of cisplatin was determined by CCK-8 method.Cell apoptosis was detected by flow cytometry.Human NK cells were isolated by magnetic activated cell sorting(MACS).Flow cytometry was used to evaluate the purity of the isolated NK cells.The cytotoxic effect of NK cells against C13K cells was assessed with LDH releasing assay.The level of IL-28A in the supernatant,the mRNA and protein expressions of IL-28A,IL-10R2 and MICA were measured by using enzyme linked immunosorbent assay,real time-PCR,Western blot and confocal spectral microscopy,respectively.Results:The expression of IL-28A mRNA in ovarian carcinoma tissues and cells was significantly lower than that in normal ovarian epithelial tissues and cells(P<0.01).The purity of NK cells was more than 80%.The IC50 of cisplatin in C13K cells was decreased by IL-28A pretreat.After pretreatment with IL-28A,the cell proliferation was inhibited,the apoptosis rate was(20.98±2.3)%,the cytotoxic effects of NK cells against C13K cells were increased.With the enhanced of intracellular IL-28A and its receptor IL-10R2 in mRNA and protein levels,the expression of MICA elevated in both mRNA and protein levels significantly(P<0.01).And IL-28A levels in the supernatant increased significantly(P<0.01).Conclusion:IL-28A can synergistically increase the cytotoxicity of cisplatin in ovarian carcinoma cells.The mechanism may be related to the inhibition of cell proliferation,induction of apoptosis,promotion of MICA expression of C13K cells,and enhancement of NK cells killing of C13K cells.