| DNMT1高表达在MNNG诱导哈萨克族食管上皮细胞恶性转化中的作用 |
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| 引用本文： | 徐漪,李旭峰,张慧霞,陈艳. DNMT1高表达在MNNG诱导哈萨克族食管上皮细胞恶性转化中的作用[J]. 癌变．畸变．突变, 2017, 29(6): 460-466. DOI: 10.3969/j.issn.1004-616x.2017.06.011 |
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| 作者姓名： | 徐漪 李旭峰 张慧霞 陈艳 |
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| 作者单位： | 新疆医科大学公共卫生学院卫生毒理学教研室,新疆乌鲁木齐,830011;新疆医科大学公共卫生学院卫生毒理学教研室,新疆乌鲁木齐830011;嘉兴学院医学院,浙江嘉兴314000 |
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| 基金项目： | 国家自然科学基金资助项目，自治区科技厅自然科学基金项目，新疆医科大学研究生创新项目 |
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| 摘 要： | 目的：利用已构建的哈萨克族食管上皮DNMT1高表达细胞模型，探讨DNMT1高表达在MNNG诱导食管上皮细胞恶性转化中的作用，为研究食管癌发病机制提供理论依据。方法：以MNNG为诱导剂，MTT法和克隆形成实验确定MNNG的恶性转化剂量；对哈萨克族食管上皮DNMT1高表达细胞和正常上皮细胞采用隔代染毒的方式进行染毒，每次染毒1 h，软琼脂集落形成实验观察不同染毒和传代次数的细胞集落形成情况；裸鼠成瘤实验观察哈萨克族食管上皮DNMT1高表达细胞及其转化细胞的恶变程度。结果：MTT法和克隆形成实验发现MNNG转化浓度为1.50×10-5 mol/L，软琼脂集落形成实验发现染毒14次第27代的哈萨克族食管上皮DNMT1高表达细胞在软琼脂上有细胞集落形成，并且随着MNNG染毒剂量、次数和细胞传代次数的增加，出现细胞形态不规则等恶性转化特点，接触抑制逐渐消失，耐受性逐渐增强，生长速度逐渐增加，而正常上皮细胞未出现阳性结果，细胞固缩死亡。接种哈萨克族食管上皮DNMT1高表达细胞的裸鼠出现局部肿块，但病理鉴定仅为结构紊乱，并未出现鳞癌，而接种转化细胞的裸鼠出现肿瘤，经病理鉴定为鳞癌。结论：成功构建了哈萨克族食管上皮DNMT1高表达细胞恶性转化模型，DNMT1高表达可促进MNNG致细胞恶性转化，加快其恶变速率和恶变程度。
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| 关 键 词： | DNMT1高表达 恶性转化 MNNG 食管上皮细胞 哈萨克族 |
| 收稿时间： | 2017-03-27 |
High expression of DNMT1 in MNNG-induced malignant transformation of Kazakh esophageal epithelial cell |
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| XU Yi,LI Xufeng,ZHANG Huixia,CHEN Yan. High expression of DNMT1 in MNNG-induced malignant transformation of Kazakh esophageal epithelial cell[J]. Carcinogenesis,Teratogenesis and Mutagenesis, 2017, 29(6): 460-466. DOI: 10.3969/j.issn.1004-616x.2017.06.011 |
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| Authors: | XU Yi LI Xufeng ZHANG Huixia CHEN Yan |
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| Affiliation: | 1. Department of Toxicology Public Health College, Xinjiang Medical University, Urumqi 830011, Xinjiang;2. College of Medicine Jiaxing University, Jiaxing 314000, Zhejiang, China |
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| Abstract: | OBJECTIVE:To explore the influence of the high expression of DNMT1 on MNNG-induced malignant transformation of Kazakh esophageal epithelial cell and to provide theoretical basis for pathogenesis of esophageal cancer.METHODS:Normal esophageal cells in culture were derived from Kazakh esophagus epithelium.MTT and clonal assays were conducted to determine the malignant transformation doses of MNNG.DNMT1 overexpression cells and the normal epithelial cells were exposed to MNNG for one hour.MTT and soft agar colony forming experiment were performed to observe the formation of cell colony under different exposure conditions and different multiplying rate.Nude mouse tumorigenicity assay was conducted to detect malignantly transformed cells.RESULTS:After treatment with MNNG (1.50 × 10-5 mol/L),cells with high expression of DNMT1 were able to form colonies in soft-agar medium after the 27th generation in vitro.These cells also formed local mass in nude mice but pathological evaluation showed that the local mass had disorderly structure and without squamous cell carcinoma morphology.On the other hand,innoculation of epithelial cells with high expression of transformed cells induced tumors in nude mice which showed squamous cell carcinoma morphology.CONCLUSION:A malignant transformation model of Kazakh esophageal epithelial cell with high DNMT1 expression was successful established.In addition,the model showed that DNMT1 high expression could promote induction of malignant transformation by MNNG. |
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| Keywords: | DNMT1 expression malignant transformation MNNG esophageal epithelial cell Kazakhstan |
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