3-甲基胆蒽诱导人脐带间充质干细胞的恶性转化  

Malignant transformation of human umbilical mesenchymal stem cells induced by 3-methycholanthrene

作　　者：唐秋灵[1] 陈秋蓉[1] 陈辉[2] 陈业增 赖秀兰[1] 刘思征[1] 谢庆东[3] 贺谷雨[2] 马廉[1] 

机构地区：[1]汕头大学医学院第二附属医院儿科,515041 [2]汕头大学医学院第二附属医院转化医学中心,515041 [3]汕头大学医学院生殖医学研究中心

出　　处：《中华实用儿科临床杂志》2013年第21期1647-1651,共5页Chinese Journal of Applied Clinical Pediatrics

基　　金：广东省自然科学基金（10151503102000033）

摘　　要：目的探讨3一甲基胆蒽（MCA）诱导人脐带间充质干细胞（HUMSCs）发生恶性转化的可能性，为其临床应用前的安全性检测提供可借鉴的检测模式及探索有效的预防恶性转化的方法。方法采用组织块贴壁法获得HUMSCs，实验组用含MCA的培养基处理HUMSCs1周，然后用基础培养基继续培养；对照组用含二甲基亚砜的培养基做同样的处理。倒置显微镜观察细胞的形态差异和生长速度，将实验组的HUMSCs注入裸鼠皮下，肿瘤长出后取出肿瘤组织做病理学鉴定，并对其进行原代培养，通过细胞染色体G显带判断是否为人源性。结果9批对照组细胞传至30～60d陆续凋亡，注射裸鼠皮下未见包块形成；9批实验组细胞中8批细胞30～60d后陆续凋亡，仅1批细胞的形态、生长状态和生长速度发生变化，表现出肿瘤细胞的一些特征；将此细胞注射于裸鼠皮下，约2个月后可形成包块，病理切片（HE染色）提示包块为恶性肿瘤，结合免疫组织化学提示病理类型为肉瘤或低分化肿瘤；肿瘤细胞常染色体显示为人源性细胞。结论HUMSCs在MCA诱导下能发生恶性转化，在体外培养的过程中可能存在安全隐患，体内移植后可能存在致瘤性风险，其临床应用的安全性仍有待证实。Objective To confirm the possible malignant transformation of human umbilical mesenchymal stem cells （HUMSCs） induced by 3-methycholanthrene （MCA） ,to seek for a noval mode that can evaluate the security of its clincial application and explore effective methods to prevent HUMSCs from malignant transformation. Methods Pri- mary HUMSCs was isolated and expanded by tissue adherent culture. The test group was treated with MCA in culture medium for a week. Then the medium was changed twice weekly. The control group was cuhured in medium containing dimethyl sulphoxide. Then 2 groups were tested for malignancy by the observations of morphological change and prolif- eration data on an inverted microscope. Transformed cells were injected subcutaneously into immune-deficient mice. Tumor tissue sections were analyzed by histology and immunochemistry to identify tumor types, and karotype analysis was taken to comfirm whether the cells come form humanbeing. Results Nine dornors of HUMSCs which propagated in cuhure continuously for up to 30 -60 days ultimately undergo senescence. Whereas 1 in 9 dornors of HUMSCs treated with MCA can bypass senescence and undergo malignant transformation which gave rise to sarcoma or poor differentia- ted tumor analysised by HE staining and immunohistochemical staining. Karotype analysis confirmed the human cell ori- gin of the tumor. Conclusions HUMSCs which treated with MCA can lead to malignant transformation. It might also represent a potential source of malignancy in vivo. Because of malignant possibility of HUMSCs, its security of clinical application is still need to be verified.

关 键 词：人脐带间充质干细胞 恶性转化 3-甲基胆蒽 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]