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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 establish a map of cervical cancer HPV16 virus integration sites by DIPS-PCR and to analyze the advantages of the integration site and integration site promoter software by software. Methods Siha cell lines were cultured in vitro. The clinical samples of single HPV16 infection were amplified by PCR and the viral integration sites of Siha cell lines and single HPV16-infected clinical cervical cancer samples were detected by DIPS-PCR. The results showed that Siha cell line HPV16 integration site 13q22; cervical cancer tissue integration sites in 1,3,5,6,8,10,11,13,14,15,17,19 and X chromosome, the integration site in the Chromosome distribution is random, but more concentrated in the brittle parts of the chromosome and its vicinity. More common in the integration site 1p35.1,5p15.3,10q24,13q21 ~ q22, Xp11.4. Analysis using CBS prediction servers and BDGP Neural Network Promoter prediction software revealed the presence of a highly suspected promoter sequence at the integration site. Conclusion DIPS-PCR can effectively detect the HPV16 integration sites in cervical cancer and Siha cell lines at the DNA level. The integration site is more likely to occur at the fragile sites of chromosomes and there may be some sites that make the viral oncogenes overexpressed Promoter sequence.