定量多基因荧光原位杂交技术在卵巢癌研究中的初步探索  

Establishing the quantitative multi-gene fluorescence in situ hybridization technique in ovarian cancer research

作　　者：黄宇婷[1] 胡林萍 鞠宝辉[1] 袁卫平 程涛 郝权[1] 

机构地区：[1]天津医科大学附属肿瘤医院妇科肿瘤科天津市肿瘤防治重点实验室,天津300060 [2]中国医学科学院血液病医院血液学研究所实验血液学国家重点实验室,天津300020

出　　处：《现代妇产科进展》2013年第2期86-89,共4页Progress in Obstetrics and Gynecology

基　　金：天津市重大科技支撑计划(No:09ZCZDSF03800);科技部国际合作项目(No:2010DFB30270);天津市卫生局基金(No:2011KZ80)

摘　　要：目的:初步探索定量多基因荧光原位杂交技术(QM-FISH)在卵巢癌基因组不稳定性研究中的应用价值。方法:采用缺口平移法,制作出Spectrum Green:c-myc,Pro-moFluor-555:Rb1,PromoFluor-590:Chk2,HyPer5:p53,PromoFluor-415:BRCA1五色荧光探针。采集卵巢癌腹水标本制片后固定细胞并进行QM-FISH检测。测量平均荧光信号强度、背景信号强度及荧光信号分裂率。结果:Spectrum Green、Cy3 v1(PromoFluor-555)、TexasRed(PromoFluor-590)、Zeiss CY5 Custom(HyPer5)、Zeiss(PF415)滤光片下观察,QM-FISH荧光信号/背景信号比值分别为12.8±0.2,8.8±0.4,3.8±0.5,5.0±0.4,9.0±0.2。荧光信号分裂率分别为(8.5±1.6)%,(12.4±1.4)%,(15.3±2.6)%,(14.4±2.2)%,(11.9±2.4)%。结论:QM-FISH技术可以成功应用于卵巢癌的研究,是研究卵巢癌基因组不稳定性、确认肿瘤标本特异性等位基因失衡极其有效的工具。Objective：To establish the quantitative multi-gene fluorescence in situ hy- bridization technique in ovarian cancer research and value its potential utilization in future clin- ical investigations. Methods： Probe DNA （ c-myc, Rbl, Chk2, p53 and BRCA1 ） was labeled with Spectrum Green, PromoFluor-555, PromoFluor-590, HyPer5 and PromoFluor-415 by nick translation. Prepare 10 samples of ovarian cancer ascite cytospin samples and then performed QM-FISH. The FISH signal was collected and assessed by comparing the relative signal intensity of each fluorophore with the autofluorescence background, ratio of fluorescence signal intensity was calculated. Split signals were also counted. Results：Ratios of fluorescence signal in Spec- trum Green, Cy3 vl （ PromoFluor-555 ） ,TexasRed （ PromoFIuor-590 ）, Zeiss CY5 Custom（ Hy- PerS） and PromoFluor-415 channels were 12.8±0.2,8.8±0.4,3.8±0.5,5.0±0.4,9.0± 0.2. Split signals counted for （ 8.5±1.6 ） %, （ 12.4 ± 1.4 ） %, （ 15.3 ± 2.6 ） %, （ 14.4 ± 2.2 ） %, （ 11.9±2.4 ） %. Conclusions： QM-FISH can be successfully applied in the research of ovarian cancer. It is powerful in evaluating genome instability and discovering the allelic imbalance.

关 键 词：原位杂交 荧光 卵巢肿瘤 细胞遗传学 比较基因组杂交 基因扩增 

分 类 号：R392.33[医药卫生—免疫学]