中国组织工程研究2017，Vol.21Issue(21)：3320-3325,6.DOI:10.3969/j.issn.2095-4344.2017.21.007
荧光定量PCR检测恶性转化人脐带间充质干细胞中DNA甲基化转移酶的改变
Quantitative real-time PCR detection of DNA methylation transferase in the malignantly transformed human umbilical cord mesenchymal stem cells
摘要
Abstract
BACKGROUND:Human umbilical cord mesenchymal stem cells (hUC-MSCs) may be mutated duringin vitro culture based on the spontaneous malignant transformation of adult stem cells and tumor stem cell theory, and there may be a risk of tumorigenesis after in vivo transplantation. Therefore, to establish and perfect the in vitro safety testing procedures will actively promote the clinical application of stem cells. OBJECTIVE:To investigate the tumorigenic mechanism of hUC-MSCs and the expression level of DNA methyltransferase (DNMTs) in hUC-MSCs. METHODS:Primary hUC-MSCs were isolated and expanded by tissue adherent culture. 3-Methycholanthrene was used to cause the malignant transformation in hUC-MSCs (experimental group), followed by morphological observation and tumorigenesis experiment in nude mice. Then, the tumor tissues were obtained and identified by pathological examination and primary cell culture, and the levels of DNMTs mRNA in hUC-MSCs treated with 3-methycholanthrene and dimethyl sulfoxide (control group) were detected by real-time RT-PCR and compared. RESULTS AND CONCLUSION:hUC-MSCs treated with 3-methycholanthrene led to malignant transformation, which showed malignant growth and non-integer ploidy changes in the cell nuclei, and formed a malignant tumor in immune-deficient mice after injection. Compared with the control group, the cells in the experimental group showed higher expression of DNMTs mRNA as detected by real-time RT-PCR. To conclude, hUC-MSCs can trigger malignant transformation in the morphology and the epigenetics under certain conditions. DNMTs can be a candidate for prevention against malignant transformation of transplanted stem cells.关键词
干细胞/脐带脐血干细胞/人脐带间充质干细胞/恶性转化/DNA甲基化/DNA甲基化转移酶分类
医药卫生引用本文复制引用
陈业增,唐秋灵,陈秋蓉,赖秀蓝,邱晓燕,郑泽鑫,李伟中..荧光定量PCR检测恶性转化人脐带间充质干细胞中DNA甲基化转移酶的改变[J].中国组织工程研究,2017,21(21):3320-3325,6.基金项目
广东省医学科学技术研究基金(B2013276) the Medical Science and Technology Research Foundation of Guangdong Province, No. B2013276 （B2013276）
