NADH对二乙基亚硝胺所致L02人肝细胞株p53基因突变和c-erbB2基因表达的影响

doi:10.3969/j.issn.1004-616x.2009.05.001

癌变·畸变·突变 ›› 2009, Vol. 21 ›› Issue (5): 329-333.doi: 10.3969/j.issn.1004-616x.2009.05.001

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NADH对二乙基亚硝胺所致L02人肝细胞株p53基因突变和c-erbB2基因表达的影响

温居一1;张积仁2;李　鹏2;张　军1;王　斌1

1.海军总医院放射肿瘤科,北京 100037;2.南方医科大学珠江医院肿瘤中心,广州 510280

收稿日期:2009-02-11 修回日期:2009-03-12 出版日期:2009-09-30 发布日期:2009-09-30
通讯作者: 温居一

Effect of NADH on Mutations of p53 Gene and Expression of c-erbB2 Gene in L02 Cells

WEN Ju-yi1; ZHANG Ji-ren2; LI Peng2; ZHANG Jun1; WANG Bin1

1.Department of Radiotherapy and Oncology, The Navy General Hospital, Beijing 100037;2.Center of Oncology, Zhujiang Hospital of Southern Medical University, Guangzhou 510280,China

Received:2009-02-11 Revised:2009-03-12 Online:2009-09-30 Published:2009-09-30
Contact: WEN Ju-yi

摘要/Abstract

摘要： 背景与目的：研究NADH对二乙基亚硝胺(DENA)所致L02人肝细胞株p53基因突变和c-erbB2基因表达的影响。材料与方法：采用聚合酶链反应-单链构象多态性(PCR-SSCP)及聚合酶链反应-限制性酶切片段长度多态性(PCR-RFLP)方法检测DENA所致的L02细胞p53基因突变；Southern blot分析DENA对c-erbB2基因表达的影响；研究辅酶NADH的抗突变作用。结果： PCR-SSCP分析显示NADH保护的DL02-Ⅲ细胞和DL02-B 细胞p53 exon7突变率均较DENA致突变组降低，差异有统计学意义(P<0.01)；PCR-RFLP分析结果显示NADH降低DENA所诱发的L02细胞p53基因第7外显子249位密码子点突变率(P<0.01)。Southern blot检测结果也显示NADH可抑制DENA所致的L02 细胞c-erbB2基因的表达上调。结论：还原型辅酶NADH具有抗突变作用，可降低DENA所致的L02细胞p53基因的突变，并抑制c-erbB2基因的表达。

关键词: 还原型烟酰胺腺嘌呤二核苷酸(NADH), 抗突变作用, 二乙基亚硝胺

Abstract: BACKGROUND AND AIM: To investigate effects of NADH on mutations of p53 gene and c-erbB2 gene in L02 cells induced by diethylnitrosamine(DENA). MATERIALS AND METHODS: The anti-mutagenesis effects of NADH on DENA-induced mutations of p53 gene in L02 cells were investigated by PCR-SSCP and PCR-RFLP analysis. The effect of NADH on c-erbB2 gene amplification was assessed by Southern blotting analysis. RESULTS: PCR-SSCP analysis showed that the mutation rate of p53 exon7 in DL02-Ⅲ and DL02-B cells in NADH protection groups decreased to 33.3％(4/12)and 50％(6/12), respectively. The difference was significant(P<0.01)compared with DENA mutagenesis group. PCR-RFLP analysis showed that NADH decreased the mutation frequency of the 249th codon of p53 in L02 cells induced by DENA. Southern blotting showed that NADH down-regulated the amplification of c-erbB2 gene in L02 cells induced by DENA. CONCLUSION: Reduced coenzyme NADH had anti-mutagenesis effects and could decrease mutation of p53 and inhibit the expression of c-erbB2 in L02 cells induced by DENA.

Key words: nicotinamide adenine dinucleotide-reduced(NADH), anti-mutagenesis, diethylnitrosamine

中图分类号:

Q754

温居一, 张积仁, 李　鹏, 张　军, 王　斌. NADH对二乙基亚硝胺所致L02人肝细胞株p53基因突变和c-erbB2基因表达的影响[J]. 癌变·畸变·突变, 2009, 21(5): 329-333.

WEN Ju-yi, ZHANG Ji-ren, LI Peng, ZHANG Jun, WANG Bin. Effect of NADH on Mutations of p53 Gene and Expression of c-erbB2 Gene in L02 Cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2009, 21(5): 329-333.

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NADH对二乙基亚硝胺所致L02人肝细胞株p53基因突变和c-erbB2基因表达的影响

Effect of NADH on Mutations of p53 Gene and Expression of c-erbB2 Gene in L02 Cells

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