Wisman G B, De Jong S, Meersma G J, Helder M N, Hollema H, de Vries E G, Keith W N, van der Zee A G
Department of Gynaecology, University Hospital Groningen, The Netherlands.
Hum Pathol. 2000 Oct;31(10):1304-12. doi: 10.1053/hupa.2000.19299.
This study was performed to determine upregulation of the human telomerase RNA component (hTR) and mRNA of the catalytic subunit of telomerase (hTERT) in (pre)malignant cervical lesions, to analyze possible intralesional heterogeneity of hTR expression, and to relate hTR and hTERT mRNA levels to telomerase activity levels and human papillomavirus (HPV) typing. hTR expression was determined by in situ hybridization (ISH) on paraffin-embedded sections, obtained from patients with cervical intraepithelial neoplasia (CIN) I-III or cervical cancer and from normal controls. hTR and hTERT mRNA expression were determined by semiquantitative rt-PCR on frozen samples from the same lesions. Data on telomerase activity and HPV were obtained from a previous study. hTR expression as determined by ISH was observed in 0 of 8 normal cervices, 1 of 14 CIN I, 15 of 28 CIN II, 21 of 30 CIN III, and 16 of 18 cervical cancer specimens. In general, hybridization patterns for hTR expression were homogeneous throughout the lesion. Frequency of hTR expression was related to grade of CIN/cervical cancer (P<.001). hTR expression, as determined by rt-PCR, was detected in 8 of 8 normal cervices, 2 of 2 CIN I, 12 of 14 CIN II, 23 of 23 CIN III, and 16 of 17 cervical cancer specimens. hTERT mRNA was detected in 1 of 8 normal cervices, 1 of 2 CIN I, 5 of 14 CIN II, 14 of 23 CIN III, and 11 of 17 cervical cancer specimens. hTR as determined by rt-PCR was not related to grade of CIN/cervical cancer, whereas hTERT mRNA expression was related to grade of CIN/cervical cancer (P<.01). hTR expression, as determined by ISH and hTERT mRNA expression by rt-PCR, were related to telomerase activity levels (P<.001, P<.05, respectively) and presence of oncogenic types of HPV (both P<.05). Our data show frequent upregulation of hTR and hTERT mRNA expression in CIN lesions, which appear to occur earlier than induction of telomerase activity. The fact that semiquantitative hTERT mRNA as well as hTR levels are related to telomerase activity levels illustrates that in (pre)malignant cervical lesions upregulation of both telomerase components may be important for functional telomerase.
本研究旨在确定人端粒酶RNA组分（hTR）和端粒酶催化亚基的mRNA（hTERT）在宫颈（癌前）病变中的上调情况，分析hTR表达可能存在的病灶内异质性，并将hTR和hTERT mRNA水平与端粒酶活性水平及人乳头瘤病毒（HPV）分型相关联。通过对石蜡包埋切片进行原位杂交（ISH）来测定hTR表达，这些切片取自宫颈上皮内瘤变（CIN）I - III级患者、宫颈癌患者及正常对照者。通过对相同病变的冷冻样本进行半定量逆转录聚合酶链反应（rt-PCR）来测定hTR和hTERT mRNA表达。端粒酶活性和HPV的数据来自先前的一项研究。ISH检测显示，8例正常宫颈组织中hTR表达为0例阳性，14例CIN I级中有1例阳性，28例CIN II级中有15例阳性，30例CIN III级中有21例阳性，18例宫颈癌标本中有16例阳性。总体而言，整个病灶内hTR表达的杂交模式是均匀的。hTR表达频率与CIN/宫颈癌分级相关（P<0.001）。rt-PCR检测显示，8例正常宫颈组织中有8例检测到hTR表达，2例CIN I级中有2例阳性，14例CIN II级中有12例阳性，23例CIN III级中有23例阳性，17例宫颈癌标本中有16例阳性。hTERT mRNA在8例正常宫颈组织中有1例检测到，2例CIN I级中有1例阳性，14例CIN II级中有5例阳性，23例CIN III级中有14例阳性，17例宫颈癌标本中有11例阳性。rt-PCR检测的hTR与CIN/宫颈癌分级无关，而hTERT mRNA表达与CIN/宫颈癌分级相关（P<0.01）。ISH检测的hTR表达和rt-PCR检测的hTERT mRNA表达均与端粒酶活性水平相关（分别为P<0.001、P<0.05），且与致癌型HPV的存在相关（均为P<0.05）。我们的数据表明，CIN病变中hTR和hTERT mRNA表达频繁上调，这似乎比端粒酶活性的诱导出现得更早。半定量hTERT mRNA以及hTR水平与端粒酶活性水平相关这一事实表明，在宫颈（癌前）病变中，两种端粒酶组分的上调可能对功能性端粒酶很重要。
