Chen Qing-Yun, Bian Mei-Lu, Chen Zhi-Hua, Liu Jun
Department of Obstetrics and Gynecology, China-Japan Friendship Hospital, Beijing 100029, China.
Zhonghua Yi Xue Za Zhi. 2005 Feb 16;85(6):400-4.
To investigate the prevalence of integration of human papillomavirus 16 (HPV16) DNA into the host genome in cervical squamous intraepithelial lesions (SIL).
Multiplex PCR was used to detect the HPV/HPV16 infection and integration status of HPV16 in the surplus cells from liquid-based cytological samples from 108 patients with cervical cancer precursor lesions. Consensus primers GP5+/GP6+ were used to amplify a 150 bp long fragment in the conserved region of the HPV L1 gene so as to detect the presence pf HPV. Scion Image 4.0 electrophoresis image analysis soft was used to calculate the E2/E6 ratio so as to evaluate the episomal and integrated status of HPV16 infection: in episomal form, both targets should be equivalent, and in integrated form, E2 gene would be absent, while in mixed form of episomal/integrated mixed form, the copy number of E2 would be less than that of E6.
Sixty-two out of the 108 patients (57.41%) had HPV infection. HPV16 were found in 32 of the 108 samples (29.63%). Among the 32 cases HPV16 DNA was exclusively episomal in 15 cases (46.88%), concomitant in 13 cases (40.62%), and integrated in 4 cases (12.50%). The prevalence of integrated and/or concomitant forms of HPV-16 DNA increased with progression of cervical disease. The prevalence of integrated form was 54.55% in the patients of CIN3 type, 50.00% in CIN2 type, 28.07% in CIN1 type, and 11.54% in the inflammatory type with significant differences between any 2 groups (all P < 0.01).
HPV16 integration into the host genome is already present in some of CIN lesions. The multiplex PCR estimation of the HPV L1, HPV16 E2, E6 genes and E2/E6 ratio could be a simple method for detecting HPV/HPV16 infection and its integration status in liquid-based residual samples. It would be a helpful complementary tool for cytological screening to identify those patients at high risk of developing high-grade squamous intraepithelial lesions and cervical cancer.
研究人乳头瘤病毒16型（HPV16）DNA整合入宿主基因组在宫颈鳞状上皮内病变（SIL）中的发生率。
采用多重聚合酶链反应（PCR）检测108例宫颈癌前病变患者液基细胞学样本剩余细胞中HPV/HPV16感染及HPV16的整合状态。使用共识引物GP5+/GP6+扩增HPV L1基因保守区150 bp长的片段，以检测HPV的存在。使用Scion Image 4.0电泳图像分析软件计算E2/E6比值，以评估HPV16感染的游离和整合状态：游离形式下，两个靶点应相等；整合形式下，E2基因缺失；游离/整合混合形式下，E2的拷贝数应少于E6。
108例患者中有62例（57.41%）感染HPV。108份样本中有32份（29.63%）检测到HPV16。在这32例中，HPV16 DNA仅为游离形式的有15例（46.88%），并存形式的有13例（40.62%），整合形式的有4例（12.50%）。HPV-16 DNA整合和/或并存形式的发生率随宫颈疾病进展而增加。CIN3型患者中整合形式的发生率为54.55%，CIN2型为50.00%，CIN1型为28.07%，炎症型为11.54%，任意两组间差异均有统计学意义（均P < 0.01）。
在部分CIN病变中已存在HPV16整合入宿主基因组的情况。对HPV L1、HPV16 E2、E6基因及E2/E6比值进行多重PCR检测，可能是一种检测液基剩余样本中HPV/HPV16感染及其整合状态的简便方法。它将是细胞学筛查的一个有用补充工具，用于识别那些发生高级别鳞状上皮内病变和宫颈癌风险较高的患者。
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