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探讨IL-21修饰的脐血间充质干细胞(UMSC)对荷卵巢癌裸鼠的治疗作用。从脐血单个核细胞中培养UMSC,经流式细胞术(FCM)鉴定后用于重组体pIRES2-IL-21-EGFP转染。RT-PCR检测UMSC中IL-21的表达。荷瘤裸鼠治疗实验分为PBS、UMSC、UMSC空质粒和UMSC-IL-21四组。以肿瘤大小、荷瘤鼠生存期判断IL-21修饰的UMSC对荷瘤鼠的治疗效应。以ELISpot法检测脾细胞IFN-γ分泌、免疫荧光法检测肿瘤组织中IL-21、NKG2D的表达,并同时检测了NK细胞活性。结果显示,脐血分离的单个核细胞经三代培养后已初步符合UMSC表型,pIRES2-IL-21-EGFP转染的UMSC能表达IL-21和EGFP。UMSC-IL-21治疗组,能抑制肿瘤生长,延长荷瘤裸鼠生存期,与其他组相比,差异有明显统计学意义(P<0.01)。UMSC-IL-21组肿瘤局部能表达IL-21与NKC2D、脾细胞分泌IFN-γ能力升高,NK细胞杀伤活性增强,表明UMSC-IL-21有良好地抗裸鼠卵巢癌作用,其机制可能与表达IL-21诱导IFN-γ分泌增加,激活NK细胞活性有关。
To investigate the therapeutic effect of IL-21 modified cord blood mesenchymal stem cells (UMSC) on ovarian cancer-bearing nude mice. UMSCs were cultured from cord blood mononuclear cells and identified by flow cytometry (FCM) for recombinant pIRES2-IL-21-EGFP transfection. The expression of IL-21 in UMSCs was detected by RT-PCR. Tumor bearing nude mice treatment experiments were divided into PBS, UMSC, UMSC empty plasmid and UMSC-IL-21 four groups. The therapeutic effect of IL-21-modified UMSCs on tumor-bearing mice was evaluated by the tumor size and the survival of tumor-bearing mice. The secretion of IFN-γ in splenocytes was detected by ELISpot assay. The expression of IL-21 and NKG2D in tumor tissue was detected by immunofluorescence assay. The NK cell activity was also detected. The results showed that mononuclear cells isolated from umbilical cord blood had initially met the UMSC phenotype after three generations of culture, and UMSCs transfected with pIRES2-IL-21-EGFP could express IL-21 and EGFP. UMSC-IL-21 treatment group, can inhibit tumor growth and prolong the survival of tumor-bearing nude mice, compared with other groups, the difference was statistically significant (P <0.01). IL-21 and NKC2D can be localized in the tumor of UMSC-IL-21 group, the ability of spleen cells to secrete IFN-γ is enhanced, and the cytotoxic activity of NK cells is enhanced, indicating that UMSC-IL-21 has good anti-ovarian cancer activity in nude mice. May be associated with the expression of IL-21 induced IFN-γ secretion increased activation of NK cell activity.