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Use of hTERT and HPV E6/E7 mRNA RT-qPCR TaqMan Assays in Combination for Diagnosing High-Grade Cervical Lesions and Malignant Tumors.

hTERT和HPV E6/E7 mRNA RT-qPCR 定量分析联合应用于诊断宫颈高级别上皮内瘤变和恶性肿瘤

Wang HY,Park S,Kim S,Lee D,Kim G,Kim Y,Park KH,Lee H

Abstract

Human papillomavirus (HPV) is a major cause of cervical cancer, which is the second most common cancer in women. HPV E6 initiates degradation of cellular tumor suppressor protein p53, induces human telomerase reverse transcriptase (hTERT) activity, and then leads to progressive cervical carcinogenesis.
In this study, the CervicGen HPV RT-qDX assay (Optipharm, Osong, Republic of Korea), which detects 16 HPV high-risk subtypes (HPV 16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, and 69), and the CervicGen hTERT RT-qDX assay (Optipharm) were evaluated using 545 ThinPrep (Hologic, Bedford, MA) Papanicolaou samples.
The positivity for the HPV E6/E7 messenger RNA (mRNA) assay was 94.4%, 95.2%, 82.4%, 46.5%, 25.0%, and 1.1% in squamous cell carcinomas, high-grade squamous intraepithelial lesions (HSILs), atypical squamous cells-cannot exclude HSIL, low-grade squamous intraepithelial lesions, atypical squamous cells of undetermined significance, and normal cytology samples, respectively. Five cervical intraepithelial neoplasia grade 2+ samples were not detected by the HPV E6/E7 mRNA assay, but they exhibited positive signals in the hTERT mRNA assay. Notably, the hTERT mRNA expression level was increased in high-grade cervical lesions but was very low in all 288 normal samples.
These data suggest that the combination of HPV E6/E7 and hTERT mRNA expression levels could be used in a complementary manner in diagnosing high-grade cervical lesions and malignant tumors and might be useful as a predictive marker in monitoring low-grade cervical lesions.

摘要

子宫颈癌是妇女第二常见的癌症,而人乳头瘤病毒(HPV)是子宫颈癌的一个主要原因。HPVE6启动细胞的肿瘤抑制蛋白p53降解,诱导人端粒酶逆转录酶(hTERT)的活性,从而导致宫颈癌变。
本研究中,对545例ThinPrep(Hologic,Bedford,MA)巴氏样本,采用CervicGenHPVRT-qDX(Optipharm,Osong,韩国)和CervicGenhTERTRT-qDX(Optipharm)对16种高危亚型HPV(HPV16、18、31、33、35、39、45、51、52、53、56、58、59、66、68、69)和hTERT进行了检测。
发现在鳞状细胞癌、高级别鳞状上皮内病变(HSILs)、非典型鳞状细胞(不能排除HSIL)、低级别鳞状上皮内病变、意义未明的非典型鳞状细胞和正常细胞样本中,HPVE6/E7信使RNA(mRNA)检测阳性率分别为94.4%、95.2%、82.4%、46.5%、25.0%和1.1%。有5例CIN2的样本未能检测出HPVE6/E7mRNA,但是hTERTmRNA检测结果为阳性。在高级别宫颈病变样本中,hTERTmRNA表达水平显着上升,而在所有288例正常样本中则表达很低。
这些结果表明,HPVE6/E7和hTERTmRNA表达水平相结合在诊断高级别宫颈病变和恶性肿瘤中能够进行互补。同时,这两者相联合检测也可能在筛查低级别宫颈病变中成为一个有用的预测指标。

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