Quantitative real-time PCR detection of DNA methylation transferase in the malignantly transformed human umbilical cord mesenchymal stem cells
10.3969/j.issn.2095-4344.2017.21.007
- VernacularTitle: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
- Author:
Yezeng CHEN
;
Qiuling TANG
;
Qiurong CHEN
;
Xiulan LAI
;
Xiaoyan QIU
;
Zexin ZHENG
;
Weizhong LI
- From:
Chinese Journal of Tissue Engineering Research
2017;21(21):3320-3325
- CountryChina
- Language:Chinese
-
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.
