Gynecologic Cancer Center, Department of Obstetrics and Gynecology, Cathay General Hospital, Taipei, Taiwan.
Int J Cancer. 2010 Aug 1;127(3):622-32. doi: 10.1002/ijc.25078.
This study aimed to evaluate whether quantitation of high-risk human papillomavirus (HR-HPV) E6 messenger RNA (mRNA) can be a potential biomarker for detecting the severity of cervical lesions. HPV genotyping was performed using a modified MY11/GP6+ PCR for HPV DNA amplification, followed by HPV genotype-specific hybridization with on a gene chip. E6 type-specific PCR was used to validate multiple infections. Quantitative real-time reverse transcriptase (QRT-PCR) and real-time PCR used to measure mRNA levels and DNA viral loads of 6 HPV oncogenic types (HPV 16, 18, 31, 33, 52 and 58) in 720 liquid-based cytology samples. The HPV DNA and RNA measurements were correlated with cervical lesions diagnosed by histopathologic examination. mRNA transcripts in the 6 types HPV DNA-positive cases was lower in normal women and <CIN 1 (23%), women with CIN 1 (54%), CIN2+ (77%) and CIN3+ (80%) (p < 0.001). Geometric mean mRNA levels ranged from 24.5 (copies per 50 ng total RNA) in normal women and <CIN 1 to 210.8 in those with CIN 1, 629.0 in CIN2+ and 699.0 in CIN3+ (p < 0.0001). Trends of increasing viral mRNA with severity of histopathologic diagnosis were significant for HPV 16, 18, 52 and 58 transcripts but not for HPV 31 and 33 transcripts. However, geometric mean DNA viral loads of HPV 16, 18, 52 and 58 DNA did not significantly increase with the severity of cervical dysplasia. Therefore, quantitative HPV E6 mRNA levels of high-risk HPV types are potentially useful biomarkers for distinguishing among HPV infections, cervical precancerous lesions and cancer.
本研究旨在评估高危型人乳头瘤病毒（HPV）E6 信使 RNA（mRNA）的定量检测是否可作为检测宫颈病变严重程度的潜在生物标志物。HPV 基因分型采用改良的 MY11/GP6+PCR 进行 HPV DNA 扩增，然后用基因芯片进行 HPV 基因型特异性杂交。采用 E6 型特异性 PCR 验证多重感染。定量实时逆转录酶（QRT-PCR）和实时 PCR 用于测量 720 例液基细胞学样本中 6 种 HPV 致癌型（HPV 16、18、31、33、52 和 58）的 mRNA 水平和 DNA 病毒载量。HPV DNA 和 RNA 测量结果与组织病理学检查诊断的宫颈病变相关。在 6 种 HPV DNA 阳性病例中，正常妇女和 <CIN 1（23%）、CIN 1（54%）、CIN2+（77%）和 CIN3+（80%）妇女的 mRNA 转录物较低（p<0.001）。正常妇女和 <CIN 1 的几何平均 mRNA 水平为 24.5（50ng 总 RNA 中的拷贝数），CIN 1 为 210.8，CIN2+为 629.0，CIN3+为 699.0（p<0.0001）。HPV 16、18、52 和 58 转录物的病毒 mRNA 水平随组织病理学诊断严重程度增加的趋势显著，但 HPV 31 和 33 转录物则不显著。然而，HPV 16、18、52 和 58 DNA 的 HPV 病毒载量的几何平均水平并未随着宫颈发育不良的严重程度而显著增加。因此，高危型 HPV 定量 HPV E6 mRNA 水平可能是区分 HPV 感染、宫颈癌前病变和癌症的有用生物标志物。
