互隔交链孢霉毒素诱发人胎食管上皮组织c-Ha-ras点突变的研究  

Point-mutation on c-Ha-ras gene of human fetal esophageal epithelium induced by mycotoxins of Alternaria alternata

作　　者：董伟华[1] 郑智敏[1] 刘桂亭[1] 林达[2] 马涧泉[2] 

机构地区：[1]河南医科大学病理生理学教研室,郑州450052 [2]中山医科大学生物化学教研室

出　　处：《中国病理生理杂志》1993年第5期604-607,共4页Chinese Journal of Pathophysiology

摘　　要：体外培养的人胎儿食管上皮组织经互隔交链孢霉毒素AME或AOH短时间(4h)处理后,提取该组织高分子量DNA;同时提取未经处理的人胎儿食管上皮组织DNA(空白对照)以及食管癌组织和癌旁组织DNA。提取的高分子量DNA作为模板进行PCR反应,PCR扩增产物经HpaⅡ酶解后进行琼脂糖凝胶电泳分析结果。电泳结果显示,经HpaⅡ酶解后,空白对照组、癌组织和癌旁组织DNA的特异性扩增产物——104bp片段消失,经AME、AOH处理的人胎食管上皮组织DNA的扩增产物——104bp片段仍然存在。结果说明,AME、AOH短时间作用后,人胎食管上皮组织c-Ha-ras基因12位密码子发生了突变;Ha-ras基因突变可能是癌变过程中的“早期事件”。Human fetal esophageal epithelial tissue were cultured in vitro and treatedwith mycotoxins of Alternaria alternata (AME or AOH) for 4 h. The genomic DNAwere extracted from these tissues. Genomic DNA was isolated from normal human fetalesophageal epithelium (as blank control), DNA from malignant tissue and its adjacentnormal mucosa was obtained from esophagectomy patients. DNA was amplified with PCRreaction, using genomic DNA as templet. The PCR products was a 104bp fragment from which the 12 codon of c-Ha-ras gene was contained. The excition point of restriction en-zyme Hpe Ⅱ was located in this fragment. The PCR amplified 104bp fragment was diges-ted by Hpa Ⅱ and analysed by agarose gel electrophoresis. The results showed that the104bp fragment amplified from genomic DNA of blank control and esophagectomy patientcould be digested by Hpa Ⅱ ; but that from genomic DNA of human fetal esophagealepithelium treated by AME or AOH could not. These results indicated that a mutationhad taken place at 12-codon of c-Ha-ras gene after it was treated by AME, AOH for ashort time. The mutation of Ha-ras gene might be the early event during esophageal car-cinogenesis. The effect of AME and AOH during the onset of esophageal cancer and themolecular machanisms of the effect were worth of further study.

关 键 词：链格孢属 霉菌毒素 食管肿瘤 

分 类 号：R735.102[医药卫生—肿瘤]