白细胞介素12基因修饰骨髓间充质干细胞对卵巢癌细胞生长的影响

发布时间：2018-04-26 10:09

本文选题：卵巢肿瘤 + 骨髓　；参考：《中国组织工程研究》2017年09期

【摘要】：背景:骨髓间充质干细胞具有来源广泛、免疫原性低等优点,尤其易于导入和表达外源基因,作为抗肿瘤基因治疗的载体具有明显优越性。目的:探讨骨髓间充质干细胞负载白细胞介素12重组腺病毒对卵巢癌细胞增殖、细胞周期和细胞凋亡的影响。方法:腺病毒介导白细胞介素12基因转染骨髓间充质干细胞,RT-PCR和Western Blotting测定骨髓间充质干细胞中白细胞介素12 m RNA和蛋白的表达,ELISA法测定细胞上清液中白细胞介素12水平。将SKOV3卵巢癌细胞与白细胞介素12重组腺病毒转染的骨髓间充质干细胞上清液共培养,对照组为SKOV3细胞与未转染骨髓间充质干细胞上清液共培养,MTT法测定SKOV3细胞增殖活性,流式细胞仪检测SKOV3细胞周期和细胞凋亡率。结果与结论:(1)腺病毒介导白细胞介素12基因成功转染到骨髓间充质干细胞中,转染后细胞有白细胞介素12 m RNA和蛋白的表达,空病毒载体组和空白对照组骨髓间充质干细胞未检测到白细胞介素12表达;(2)培养48 h,白细胞介素12组骨髓间充质干细胞上清液中白细胞介素12水平为(68.78±12.35)μg/L,空病毒载体组和空白对照组骨髓间充质干细胞上清液中未检测到白细胞介素12;(3)白细胞介素12转染组骨髓间充质干细胞上清液对SKOV3细胞增殖有抑制作用,细胞增殖抑制率随时间的延长而增加(P0.05)。转染组SKOV3细胞G1期细胞比例高于对照组(P0.05),G2期细胞比例低于对照组(P0.05)。转染组SKOV3细胞凋亡率高于对照组(P0.05);(4)结果表明,转染白细胞介素12基因的骨髓间充质干细胞能够表达白细胞介素12,其培养上清液能够抑制卵巢癌细胞增殖并诱导其凋亡。
[Abstract]:Background: bone marrow mesenchymal stem cells (BMSCs) have many advantages, such as wide source, low immunogenicity and so on. Aim: to investigate the effects of bone marrow mesenchymal stem cells loaded with interleukin 12 recombinant adenovirus on the proliferation, cell cycle and apoptosis of ovarian cancer cells. Methods: the expression of interleukin 12 RNA and protein in bone marrow mesenchymal stem cells were detected by RT-PCR and Western Blotting. The levels of interleukin 12 in supernatant of bone marrow mesenchymal stem cells were determined by Elisa. SKOV3 ovarian cancer cells were co-cultured with the supernatant of bone marrow mesenchymal stem cells transfected with interleukin-12 recombinant adenovirus, while SKOV3 cells and untransfected bone marrow mesenchymal stem cells supernatants were co-cultured to determine the proliferative activity of SKOV3 cells. SKOV3 cell cycle and apoptosis rate were measured by flow cytometry. Results and conclusion adenovirus-mediated interleukin-12 gene was successfully transfected into bone marrow mesenchymal stem cells, and IL-12 RNA and protein were expressed in the transfected cells. The level of interleukin 12 in the supernatant of bone marrow mesenchymal stem cells was 68.78 卤12.35 渭 g / L after 48 h culture. The supernatant of bone marrow mesenchymal stem cells in body group and blank control group had no effect on the proliferation of SKOV3 cells in the supernatant of bone marrow mesenchymal stem cells (BMSCs), which was not detected in the supernatant of bone marrow mesenchymal stem cells (BMSCs). The inhibitory rate of cell proliferation increased with time. The percentage of SKOV3 cells in G1 phase in transfected group was higher than that in control group, which was lower than that in control group. The results showed that bone marrow mesenchymal stem cells transfected with interleukin-12 gene could express interleukin-12, and its supernatant could inhibit the proliferation and induce apoptosis of ovarian cancer cells.
【作者单位】：河北医科大学第一医院;
【基金】：河北省2014年度医学科学研究重点课题计划项目(ZD20140325)~~
【分类号】：R737.31

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