摘要
背景与目的:VHL基因突变与肾透明细胞癌发生关系密切,而CyclinD1基因通过促进细胞过度增殖,对肾透明细胞癌发生和发展起重要作用。本研究拟初步探讨肾透明细胞癌中VHL基因突变的状况及其与CyclinD1表达上升的相关性。方法:取50例肾透明细胞癌手术标本,抽提病变组织和癌周正常组织中的基因组DNA和总RNA。通过PCR扩增VHL基因各外显子序列并测序以及内切酶鉴定的方法,确定肿瘤组织中VHL基因突变的比率和具体位点。通过RT-PCR和Westernblot方法,检测相应肿瘤组织中CyclinD1的表达水平是否发生改变。结果:VHL基因序列检测结果表明,有42例(84.0%)肾透明细胞癌标本的VHL基因发生各种形式的突变,有12(24.0%)例甚至发生2种以上的基因突变。在57例VHL基因外显子突变中,1号外显子突变有17例(29.8%);2号外显子突变有26例(45.6%);3号外显子突变有14例(24.6%)。CyclinD1的表达水平在42例VHL基因发生突变的组织中均有不同程度的上升,达到正常对照组的2～10倍(3.91±1.54倍)(P<0.01),另8例表现为正常。结论:在肾透明细胞癌中存在各种VHL基因突变,导致CyclinD1过度表达。
BACKGROUND ＆ OBJECTIVE. The mutation of von Hippel- Lindau （VHL） tumor suppressor gene is closely related with tumorigenesis of renal clear cell carcinoma （RCCC）. Cyclin D1 gene plays an important role in progression of RCCC by stimulating cell proliferation. This study was to determine the mutation of VHL gene in RCCC, and explore its correlation to overexpression of Cyclin D1. METHODS： The specimens of RCCC and adjacent normal renal tissue from 50 patients were collected after surgery. Total RNA and genomic DNA were extracted from each sample. Variant exons in VHL gene were amplified by polymerase chain reaction （PCR） and sequenced, and DNA hypermethylation was detected by restriction analysis. Reverse transcription-PCR （RT-PCR） and Western blot were used to detect the expression of Cyclin D1. RESULTS. Of the 50 specimens, 42 （84.0%） had various VHL gene mutations, 12 （24.0%） had more than 1 kind of gene mutation. Of the 57 cases of exon mutation of VHL gene, 17 （29.8%） were located in exon 1, 26 （45.6%） in exon 2, and 14 （24.6%） in exon 3. The expression of Cyclin D1 in the 42 cases with VHL gene mutation was increased to 2-10 （3.91±1.54） times that of normal controls （P〈0.01）. Cyclin D1 expression in the other 8 cases was normal. CONCLUSION： There are variant mutations of VHL gene in RCCC, which may lead to overexpression of Cyclin D1.
出处
《癌症》
SCIE
CAS
CSCD
北大核心
2006年第2期175-178,共4页
Chinese Journal of Cancer
