Jiang Runqiu, Xu Wenrong, Zhu Wei, Chen Miao, Qian Hui, Qiao Chun, Yang Huan, Wang Xingzhong, Chen Yongchang
School of Medical Technology, Zhenjiang Key Institute of Clinical Laboratory Medicine, Jiangsu 212001, PR China.
Oncol Rep. 2006 Nov;16(5):1021-8.
In previous experiments, a novel tumor cell line, which was characterized by dominated F6 mutated from human mesenchymal stem cells (hMSCs), was developed. The mechanism and biological characteristics of this mutation are still unclear. In this study, the histological type of F6 cells was investigated by immunohistochemistry with specific markers: vimentin, CD117, desmin, NSE and vWF. The characteristics of proliferation and metastasis were shown by PCNA (proliferating cell nuclear antigen), and nm23 and cell cycle-related genes, such as p16, p21, p53 and pRb, were analyzed by RT-PCR and immunohistochemistry. The expression of hTRAP and BMI-1 were detected by real-time PCR and Western blotting. The activity of telomerase was analyzed by TRAP (telomerase repeat amplification protocol) assay. The results showed that multi-directional differentiation occurred in F6 cells, i.e., special markers of muscle, endothelial cell and nerve system were co-expressed in F6 cells, while hardly expressed in hMSCs. F6 cells maintained the same properties as of MSCs, such as negativity for both CD117 and vimentin. F6 cells exhibited strong positivity for PCNA and negativity for nm23. The cell cycle-related genes, such as p16, p21, p53 and pRb, were not detected in F6 cells, while the expression of hTRAP and BMI-1 was significantly higher. The activity of telomerase was also significantly higher in F6 cells than that in hMSCs. These findings indicated that multi-directional differentiation occurred during the transformation of hMSCs into F6 cells, and that the genes of cell cycle and cell senescence may also be associated with the neoplasia of adult stem cells.
在先前的实验中，开发了一种新型肿瘤细胞系，其特征是源自人间充质干细胞（hMSCs）的F6占主导地位且发生了突变。这种突变的机制和生物学特性仍不清楚。在本研究中，通过使用特异性标志物波形蛋白、CD117、结蛋白、神经元特异性烯醇化酶和血管性血友病因子进行免疫组织化学研究F6细胞的组织学类型。通过增殖细胞核抗原（PCNA）显示增殖和转移特性，并通过逆转录聚合酶链反应（RT-PCR）和免疫组织化学分析nm23以及细胞周期相关基因，如p16、p21、p53和视网膜母细胞瘤蛋白（pRb）。通过实时PCR和蛋白质印迹法检测人端粒酶逆转录酶（hTRAP）和BMI-1的表达。通过端粒酶重复序列扩增法（TRAP）分析端粒酶活性。结果表明，F6细胞发生了多向分化，即肌肉、内皮细胞和神经系统的特异性标志物在F6细胞中共表达，而在hMSCs中几乎不表达。F6细胞保持了与间充质干细胞相同的特性，如对CD117和波形蛋白均呈阴性。F6细胞对PCNA呈强阳性，对nm23呈阴性。在F6细胞中未检测到细胞周期相关基因，如p16、p21、p53和pRb，而hTRAP和BMI-1的表达明显更高。F6细胞中端粒酶活性也明显高于hMSCs。这些发现表明，在hMSCs向F6细胞转化过程中发生了多向分化，并且细胞周期和细胞衰老基因可能也与成体干细胞的肿瘤形成有关。
