宫颈癌组织和Siha细胞株HPV16病毒整合位点的检测与分析

Detection and Analysis the HPV16 Integration in the Specimens of the Cervical Carcinoma and the Cell Line Siha

目的利用DIPS-PCR方法建立宫颈癌组织HPV16病毒整合位点图谱,并利用软件分析存在的优势整合位点及整合位点处启动子序列。方法细胞培养Siha细胞株,PCR扩增并筛选出单一HPV16感染的临床标本,利用DIPS-PCR方法检测Siha细胞株和单一HPV16感染的临床宫颈癌标本中病毒整合位点。结果发现Siha细胞株HPV16整合位点13q22;宫颈癌组织整合位点在1、3、5、6、8、10、11、13、14、15、17、19及X染色体上,整合位点在染色体的分布是随机的,但其多集中于染色体脆性部位及其附近区域。整合位点多见于1p35.1、5p15.3、10q24、13q21~q22、Xp11.4。利用CBS prediction servers和BDGP Neural Network Promoter prediction软件分析发现整合位点存在高度疑似启动子序列。结论 DIPS-PCR在DNA水平可有效检测宫颈癌组织和Siha细胞株HPV16整合位点,整合位点多出现在染色体的脆性位点处,且该脆性部位可能存在某些使病毒癌基因过度表达的启动子序列。

Objective To build the viral integration sites map and verify the integration sites at or near the fragile site.Methods PCR method were applied to screen single HPV16 infected cervical carcinoma specimen and the viral integration sites in the specimens of cervical carcinomas and the cell line cells were detected by detection of integrated papillomavirus seguences by ligation-mediated PCR（DIPS-PCR）.Results The integration site in cell line Siha was 13q22.The integration sites in the specimens of the cervical cancinomas were randomly located in chromosome 1,3,5,6,8,10,11,13,14,15,17,19 as well as in X,but the sites were most likely at or near the chromosome fragile site.The location of the integration sites were at 1p35.1,5p15.3,10q24,13q21-q22 and Xp11.4.By using the softwares CBS prediction servers and BDGP Neural Network Promoter prediction we found that the integration sites were located at the promoter like sequencs.Conclusion DIPS-PCR can detect the viral integration sites,and the sites are at or near the chromosome fragile sites,and the integration sites may including the promoter to amplify the virus oncogene.