Janušicová V, Kapustová I, Zúbor P, Slezák P, Kajo K, Lasabová Z, Plank L, Danko J
Gynekologicko-porodnicka klinika, Martin.
Ceska Gynekol. 2012 Jun;77(3):245-50.
To determine the presence of HPV infection and expression level of p16INK4A mRNA transcripts in cervical smears as adjunct biomarker in detection of cervical intraepithelial neoplasia or cancer.
Prospective pilot clinical study assessing clinical utility and validity of ddCt method for qPCR mRNA expression of p16ink4a in comparison to immunohistochemistry.
Department of Molecular Biology, Department of Obstetrics and Gynecology, Jessenius Medical Faculty, Commenius University, Martin, Slovak Republic.
Cervical smears (OC) from patients with different cervical lesions (L-SIL, H-SIL, SCA; n=45) and from healthy controls (n=45) were tested for the presence of HPV infection and p16INK4A mRNA transcripts using relative quantification (RQ). Results were compared to H&E and IHC histological findings from biopsies (conization, hysterectomy).
HPV 16 was the most frequent finding (53.3%) in the group of subjects with cervical dysplasia. The p16INK4A mRNA expression analysis revealed the slightly reduced expression in L-SIL group, 4-fold increased expression in H-SIL and 10-fold increase in women with SCA when compared to controls. The p16INK4A mRNA expression in OC was present in 30% of L-SIL, 75% of H-SIL and 85.7% of SCA samples, respectively. The test overall sensitivity was 81.48% (95% CI: 61.92-93.7) and specificity 60% (95% CI: 26.24- 87.84) with PPV of 84.62% and NPV of 54.55%. The likelihood ratio (LR) in case of test positivity was 2.04 and for negativity 0.31. The diagnostic accuracy of p16INK4A expression by RQ method in OC smears for prediction of p16 positivity in cervical dysplasia was 66.7% for the L-SIL lesions, 59.5% for H-SIL lesions, and 100% for SCA (r=0.9897, p<0.0913) when compared to IHC p16 positive findings in surgically treated samples.
The relative quantification is able to determine the level of p16INK4A mRNA transcripts in cervical smear cells with active carcinogenesis nearly at the same level as IHC staining. The advance of biopsy sparing over IHC is qualifying this diagnostic approach for useful candidate in selective management of women with cervical dysplasia looking for cervix preservation or avoiding the unnecessary overtreatment.
确定宫颈涂片中人乳头瘤病毒（HPV）感染情况及p16INK4A mRNA转录本的表达水平，作为检测宫颈上皮内瘤变或癌症的辅助生物标志物。
前瞻性试点临床研究，评估与免疫组化相比，ddCt法用于p16ink4a的qPCR mRNA表达的临床实用性和有效性。
斯洛伐克共和国马丁市康梅纽斯大学耶塞纽斯医学院分子生物学系、妇产科系。
采用相对定量（RQ）法检测不同宫颈病变（低度鳞状上皮内病变、高度鳞状上皮内病变、鳞状细胞癌；n = 45）患者及健康对照者（n = 45）的宫颈涂片（OC）中的HPV感染情况和p16INK4A mRNA转录本。将结果与活检（锥切术、子宫切除术）的苏木精-伊红染色（H&E）和免疫组化（IHC）组织学结果进行比较。
在宫颈发育异常组中，HPV 16是最常见的发现（53.3%）。与对照组相比，p16INK4A mRNA表达分析显示，低度鳞状上皮内病变组表达略有降低，高度鳞状上皮内病变组表达增加4倍，鳞状细胞癌女性表达增加10倍。在OC中，p16INK4A mRNA表达分别出现在30%的低度鳞状上皮内病变、75%的高度鳞状上皮内病变和85.7%的鳞状细胞癌样本中。该检测总体灵敏度为81.48%（95%置信区间：61.92 - 93.7），特异性为60%（95%置信区间：26.24 - 87.84），阳性预测值为84.62%，阴性预测值为54.55%。检测阳性时的似然比（LR）为2.04，阴性时为0.31。与手术治疗样本中免疫组化p16阳性结果相比，RQ法检测OC涂片p16INK4A表达对宫颈发育异常中p16阳性的预测诊断准确性在低度鳞状上皮内病变中为66.7%，高度鳞状上皮内病变中为59.5%，鳞状细胞癌中为100%（r = 0.9897，p < 0.0913）。
相对定量能够确定宫颈涂片细胞中p16INK4A mRNA转录本的水平，在活跃致癌过程中其水平与免疫组化染色几乎相同。与免疫组化相比，活检保留的优势使这种诊断方法成为寻求保留宫颈或避免不必要过度治疗的宫颈发育异常女性选择性管理中有价值的候选方法。
