白细胞介素-12基因修饰人脐带间充质干细胞抗裸鼠移植瘤的作用

Inhibitory effect of AdIL-12-infected umbilical cord mesenchymal stem cells on xenograft in nude mice

目的观察以人脐带间充质干细胞（UC—MSCs）为基因治疗载体表达白细胞介素-12（IL-12）抗裸鼠移植瘤的作用。方法腺病毒介导IL-12基因转染UC—MSCs（AdIL-12-MSCs），Westernblot和逆转录．聚合酶链反应（RT—PCR）检测IL-12基因蛋白及mRNA表达，流式细胞仪检测其对卵巢癌SKOV3细胞周期影响；建立裸鼠移植瘤模型，按组别行尾静脉移植，计算抑瘤率、凋亡率，免疫组织化学检测移植瘤血管内皮生长因子（VEGF）和毛细血管密度（MVD）表达。结果AdIL-12-MSCs在蛋白及mRNA水平表达IL-12基因，抑制SKOV3细胞增殖；移植瘤UC—MSCs组、Ad—MSCs组、AdIL．12组、AdIL-12-MSCs组、环磷酰胺组、联合组抑瘤率为10．06％、8．25％、50．91％、58．23％、70．43％、81．10％，凋亡率为（9．37±0．09）％、（8．96±0．15）％、（15．62±1．22）％、（18．27±1．49）％、（22．07±0．23）％、（25．28±2．05）％；AdIL-12-MSCs能够降低移植瘤VEGF及MVD表达。结论AdlL-12-MSCs能够在蛋白及mRNA水平表达IL-12基因，抑制SKOV3细胞增殖，抑制SKOV3移植瘤生长和血管生成，与化疗药物联用抑瘤作用增强。

Objective To study the influence of umbilical cord mesenchymal stern cells （UC-MSCs） infected with AdlL-12 on xenograft in nude mice. Methods Recombinant adenovirns vectors harboring interleukin （IL）-12 were infected into UC-MSCs to construct AdlL-12-MSCs. Western blotting and reverse transcription-polymerase chain reaction （RT-PCR） were used to detect IL-12 protein and mRNA expression. Flow cytometry was used to detect the effect of AdlL-12-MSCs on the cell cycles of SKOV3 cells. The model of ovarian carcinoma SKOV3 transplanted subcutaneously was established in nude mice and the grafting was done via tail vein. The inhibition rate and apoptosis rate was calculated, the expression of vascular endothelial growth factor （VEGF） and micmvascular density （MVD） was detected by using im- munohistochemistry. Results IL-12 was expressed in UC-MSCs at both protein and mRNA levels. The supernatant of AdIL-12-MSCs inhibited the proliferation of SKOV3 cells. The inhibition rate of UC-MSCs group, Ad-MSCs group, AdIL-12 group, AdIL-12-MSCs group, CTX group and combined group （AdIL-12-MSCs ± CTX） was 10.06% , 8.23% , 50. 91% , 58.23% , 70. 43% , 81.10% , and apoptosis ratewas （9.37±0.09）%, （8.96±0.15）%, （15.62±1.22）%, （18.27 ±1.49）%, （22.07± 0. 23 ） % , （25.28 ± 2.05） % , respectively. Immunohistoehemistry revealed that AdlL-12-MSCs could reduce the expression of VEGF and MVD. Conchision AdlL-12-MSCs can express IL-12 at protein and mRNA levels, inhibit the proliferation of SKOV3 cells, inhibit the growth and angiogenesis of ovarian carcinoma SKOV3 transplanted subcutaneously in nude mice. AdlL-12-MSCs in combination with chemotherapy drugs could enhance the antitumor effect.