Raji细胞中EB病毒染色体整合的荧光原位杂交定位  

Identification of EBV chromosomal integration sites in Raji cells by fluorescence in situ hybridization

作　　者：高建明[1] 李小玲[2] 李桂源[2] 

机构地区：[1]三峡大学医学院病理生理学教研室,湖北宜昌443002 [2]中南大学肿瘤研究所,长沙410078

出　　处：《中南大学学报（医学版）》2009年第1期13-19,共7页Journal of Central South University ：Medical Science

基　　金：国家自然科学基金重点项目(30330560)~~

摘　　要：目的：对Raji细胞中EBV整合位点进行染色体定位。方法：用Southern免疫印迹检测Raji细胞中EBV DNA，应用G显带和荧光原位杂交（fluorescence in situ hybridization，FISH）对Raji细胞中EBV整合位点进行染色体定位。结果：Raji细胞gDNA经BamHI酶切消化后，与同位素标记的Probe-1（EBV基因组13232-16189）杂交，阳性片段大小为10kb和4kb，与Probe-2（EBV基因组5～3271）杂交，阳性片段大小为23kb。Raji细胞中EBV整合位点有1p，1q，2q，3p，3q，4q，5q，6q，7p，7q，9q，11p，14q，15q等，其中4q，2q，1q，7q为EBV DNA整合的高频位点。计数33个整合信号，分别有7，4，4，4个信号位于染色体4q，2q，1q，7q，这4个位点的信号占所有信号的构成比为64％，15号以后的染色体及2条性染色体均未见EBV整合。结论：本研究用G显带和FISH方法首次对Raji细胞中EBV整合位点进行了定位，Raji细胞中EBV整合具有一些高频位点，是一种非随机性整合。Objective To identify the Epstein-Barr virus （EBV） chromosomal integration sites in Raji cells. Methods EBV DNA was detected by Southern hybridization, and the viral chromosomal integration sites were identified using G banding and fluorescence in situ hybridization （FISH）. Results BamHI-digested genomie DNA from Raji cells was hybridized with ^32p-labeled probe-1 （EBV genome 13 232 -16 189） and Probe-2 （EBV genome 5 -3 271 ） , which generated 4 and 10, 23 kb positive bands respestively. The viral integration sites included 1 p, 1 q, 2q, 3p, 3q, 4q, 5q, 6q, 7p, 7q, 9q,11p, 14q, and 15q,and chromosomal bands 4q, 2q, 1q and 7 q were viral integration sites with high frequencies. Among the 33 signals counted, 7, 4, 4, and 4 signals were at the site 4 q, 2 q, 1 q, and 7 q respectively, and 64 % of the total signals were found in these 4 chromosomal bands. No viral integration occurred in chromosomes 16 - 22 or the sex chromosomes （X, Y ）. Conclusion This study firstly identifies the EBV integration sites in Raji cells using G banding and FISH. There are some viral integration sites with high frequencies in Raji cells, and EBV integrates into Raji cell genomes non-randomly.

关 键 词：RAJI细胞 EB病毒 整合 荧光原位杂交 

分 类 号：R73-3[医药卫生—肿瘤]