应用基因芯片诊断宫颈石蜡组织样本中人乳头状瘤病毒感染及其临床意义

Diagnosis of human papillomavirus infection in paraffin-embedded cervical tissues by gene clip technology

目的探讨应用基因芯片检测宫颈石蜡组织标本中人乳头状瘤病毒(HPV)感染的可能性及其临床意义。方法收集解放军总医院诊断为宫颈鳞状上皮病变的石蜡组织标本40例,其中宫颈浸润性鳞癌18例,宫颈上皮内瘤变(CIN)Ⅲ12例,CINⅠ4例,CINⅡ6例。从组织中提取DNA后采用基因芯片检测23种常见HPV基因亚型,即PCR扩增后产物在基因芯片上进行杂交。同时选用10例经基因芯片检测16型和18型基因阳性的宫颈鳞癌的石蜡组织切片做原位杂交。基因芯片检测结果与部分原位杂交结果进行比较并分析。结果基因芯片检测的18例宫颈鳞癌HPV高危亚型均为阳性(100%),其中1例为混合阳性;12例CINⅢ中11例为高危亚型阳性(91·7%),1例阴性;6例CINⅡ的宫颈病变中高危型5例阳性,低危型1例阳性;4例CINⅠ中有2例低危型阳性、2例阴性;宫颈鳞癌和CINⅢ组与CINⅠ和Ⅱ组比较,差异有统计学意义(U=80·0,P<0·01)。10例宫颈鳞癌基因芯片HPV16型和18型阳性组织中,原位杂交同型探针6例检测显示阳性。结论HPV基因芯片技术可用于检测多种亚型,特异性强,敏感性高,对HPV感染亚型的鉴别及宫颈癌的预防和治疗具有重要意义。

Objective To study the feasibility of diagnosing of human papillomavirus （HPV） infection in paraffin-embedded cervical tissues by high-throughput gene chip technology and its clinical significance. Methods Forty cases of HPV-related cervical lesions, including 18 cases of invasive squamous cell carcinoma, 12 cases of cervical intraepithelial neoplasia （CIN） Ⅲ, 6 cases of CIN Ⅱ and 4 cases of CIN Ⅰ , were enrolled. DNA was extracted from paraffin-embedded tissues and amplified by polymerase chain reaction （PCR） using HPV DNA primers. The PCR products were then reversely hybridized with gene chip technology. The results were compared with that of in-situ hybridization （ISH）. Results All of the 18 cases of cervical squamous cell carcinoma were positive for high-risk HPV genotypes （with 1 case showing a mixture with low-risk genotypes）. In contrast,11 cases （91.7%） of CIN Ⅲ, 5 cases （83%） of CIN Ⅱ and none of the CIN Ⅰ cases were positive for high-risk HPV genotypes. On the other hand, low-risk HPV genotypes were detected only in 1 case （ 17% ） of CIN Ⅱ and 2 cases （50%） of CIN Ⅰ. The difference between the two groups （ CIN Ⅲ/squamous cell carcinoma versus CIN Ⅰ / CINⅡ ） was statistically significant （ U = 80. 0, P 〈 0. 01 ）. Among the 10 squamous carcinoma cases positive for HPV types 16 and 18 by gene chip technology, high-risk HPV DNA was also detected in 6 of them when using in-situ hybridization. Conclusions Gene chip technology is able to detect multiple HPV genotypes in paraffln-embedded tissues with high sensitivity and specificity. The distinction between low and high-risk HPV genotypes is seemed useful in prevention and management of cervical cancer.