山东青岛地区宫颈癌组织中ＨＰＶ１６型Ｅ６和Ｅ２基因突变分析

目的　通过分析山东青岛地区宫颈组织中ＨＰＶ１６型Ｅ６和Ｅ２基因突变情况，探讨其与该地区宫颈癌的关系。方法　从１０４例青岛地区宫颈疾病组织中提取ＤＮＡ作为模板，采用聚合酶链式反应（ＰＣＲ）技术筛选出高危型ＨＰＶ及ＨＰＶ１６阳性标本，扩增出ＨＰＶ１６型Ｅ６、Ｅ２全长基因，ＰＣＲ产物纯化后测序，与德国ＨＰＶ标准株进行比对分析。结果　宫颈组织中高危型ＨＰＶ阳性率为９３.２７％（９７／１０４），ＨＰＶ１６阳性率为６９.２３％（７２／１０４）。ＨＰＶ１６阳性标本中扩增出Ｅ６基因３７例，有５例与标准株序列相同，３２例存在突变，其中２５例突变型别为Ｔ１７８Ｇ或Ｔ１７８Ａ（Ｄ２５Ｅ）。在Ｅ２基因全序列测序中，２３例均存在Ｃ３６８４Ａ（Ｔ-Ｋ），１４例同时存在Ｔ３５２４Ｃ、Ｃ３６８４Ａ（Ｔ-Ｋ）和Ｃ３７８７Ａ（Ｄ-Ｅ），９例同时存在Ａ２９２６Ｇ、Ｃ３１５９Ａ（Ｔ-Ｋ）、Ｇ３２４９Ａ（Ｒ-Ｑ）、Ｔ３３８４Ｃ（Ｉ-T）、Ｃ３４１０Ｔ（Ｐ-Ｓ）和Ｃ３６８４Ａ（Ｔ-Ｋ）。结论　山东青岛地区宫颈癌患者ＨＰＶ１６型Ｅ６、Ｅ２基因与德国标准株比较存在多处变异，Ｅ６与Ｅ２基因突变可能存在相关性。

Mutations of E6 and E2 oncogenes of human papillomavirus 16 in women with cervical carcinoma in Qingdao

Objective To investigate the distribution of HPV16 E6 and E2 gene mutation in cervical cancer, and the relationship between the mutation and the development of cervical carcinoma in Qingdao, Shandong. Methods DNA samples extracted from 104 cervical cancer tissues in Qingdao were amplified by polymerase chain reaction （PCR） using specific primers for high-risk HPV, HPV16, E6 and E2 genes of HPV16, respectively. Then the PCR fragments of HPV16 E6 and E2 genes were purified, sequenced and compared with the type stain. Results The positive rates of HPV and HPV16 were 93.27% （97/104） and 69. 23% （72/ 104） respectively. Prototype HPV16 E6 was found in 5 cases （ 13.51% ） while mutation was detected in 32 cases （86. 49% ）, among which the T178G or T178A （D25E） mutation was found in 25 cases （67.57 % ）. The E2 sequence of 23 cases contained C3684A （ T- K）, that of 14 eases contained T3524C, C3684A （T-K） and C3787A （D-E） at the same sample, and 9 eases contained A2926G, C3159A（T-K）, G3249A（R-Q）, T3384C（I-T）, C3410T（P-S） and C3684A（T-K） meanwhile. Conclusion There are several HPV16 E6 and E2 gene mutations in cervical eancer tissue for women in Qingdao. The mutation of E6 may relate to that of E2.