【摘要】：背景:根據(jù)體外培養(yǎng)的成體干細(xì)胞自發(fā)惡性轉(zhuǎn)化現(xiàn)象及腫瘤干細(xì)胞理論,人臍帶間充質(zhì)干細(xì)胞在體外培養(yǎng)過(guò)程中可能產(chǎn)生變異,體內(nèi)移植后可能存在致瘤性風(fēng)險(xiǎn)。因此,建立健全的體外安全性檢測(cè)程序,將在干細(xì)胞臨床應(yīng)用上起積極推動(dòng)作用。目的:探討人臍帶間充質(zhì)干細(xì)胞致瘤性機(jī)制及DNA甲基化轉(zhuǎn)移酶的表達(dá)水平。方法:采用組織塊貼壁法分離、培養(yǎng)、擴(kuò)增出人臍帶間充質(zhì)干細(xì)胞,利用3-甲基膽蒽促使人臍帶間充質(zhì)干細(xì)胞發(fā)生惡性轉(zhuǎn)化,通過(guò)形態(tài)學(xué)觀察,裸鼠成瘤實(shí)驗(yàn)進(jìn)行驗(yàn)證,成瘤組織進(jìn)行病理學(xué)鑒定、原代細(xì)胞培養(yǎng),熒光定量PCR檢測(cè)人臍帶間充質(zhì)干細(xì)胞與腫瘤細(xì)胞DNA甲基化轉(zhuǎn)移酶表達(dá)水平的差異。結(jié)果與結(jié)論:(1)人臍帶間充質(zhì)干細(xì)胞經(jīng)3-甲基膽蒽處理后形成惡性轉(zhuǎn)化細(xì)胞,呈惡性生長(zhǎng),細(xì)胞核型呈非整數(shù)倍體改變,注入裸鼠體內(nèi)后能形成惡性腫物;(2)經(jīng)熒光定量PCR檢測(cè),惡性轉(zhuǎn)化后腫瘤細(xì)胞(實(shí)驗(yàn)組)相比二甲基亞砜培養(yǎng)人臍帶間充質(zhì)干細(xì)胞(對(duì)照組),其DNA甲基化轉(zhuǎn)移酶的基因表達(dá)量明顯增高;(3)結(jié)果表明,人臍帶間充質(zhì)干細(xì)胞在一定條件下能發(fā)生惡性轉(zhuǎn)化,包括形態(tài)學(xué)變化及表觀遺傳學(xué)層面改變,DNA甲基化轉(zhuǎn)移酶可作為預(yù)防干細(xì)胞移植后成瘤的進(jìn)一步研究方向。
[Abstract]:Background: according to the spontaneous malignant transformation of adult stem cells cultured in vitro and the theory of tumor stem cells, human umbilical cord mesenchymal stem cells may mutate in vitro, and there may be tumorigenic risk after in vivo transplantation. Therefore, the establishment of a sound safety detection procedure in vitro will play an active role in the clinical application of stem cells. Objective: to investigate the tumorigenic mechanism of human umbilical cord mesenchymal stem cells and the expression of DNA methyltransferase. Methods: human umbilical cord mesenchymal stem cells were isolated, cultured and expanded by tissue block adherent method. 3-methylcholanthracene was used to promote the malignant transformation of human umbilical cord mesenchymal stem cells, which were verified by morphological observation and nude mice tumorigenesis experiment. Pathological identification of tumorigenic tissues, primary cell culture and fluorescence quantitative PCR were used to detect the expression of DNA methyltransferase between human umbilical cord mesenchymal stem cells and tumor cells. Results and conclusion: (1) Human umbilical cord mesenchymal stem cells were treated with 3-methylcholanthracene to form malignant transformed cells with malignant growth and aninteger ploidy change in karyotype. Malignant masses could be formed after injection into nude mice. (2) the expression of DNA methyltransferase gene in malignant transformed tumor cells (experimental group) was significantly higher than that in human umbilical cord mesenchymal stem cells cultured with dimethyl sulfoxide (control group) by fluorescence quantitative PCR. (3) the results showed that human umbilical cord mesenchymal stem cells could undergo malignant transformation under certain conditions, including morphological changes and epigenetic changes. DNA methyltransferase can be used as a further research direction to prevent tumor formation after stem cell transplantation.
【作者單位】： 汕頭大學(xué)醫(yī)學(xué)院第二附屬醫(yī)院兒科;汕頭大學(xué)醫(yī)學(xué)院第二附屬醫(yī)院轉(zhuǎn)化醫(yī)學(xué)中心;
【基金】：廣東省醫(yī)學(xué)科學(xué)技術(shù)研究基金(B2013276)~~
【分類號(hào)】：R457.7

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