Migdalska-Sęk Monika, Karowicz-Bilińska Agata, Pastuszak-Lewandoska Dorota, Czarnecka Karolina H, Nawrot Ewa, Domańska-Senderowska Daria, Kiszałkiewicz Justyna, Brzeziańska-Lasota Ewa
Department of Molecular Bases of Medicine, 1st Chair of Internal Diseases, Medical University of Lodz, Pomorska St. No. 251, 92-213, Lodz, Poland.
Department of Pregnancy Pathology, 1st Chair of Gynecology and Obstetrics, Medical University of Lodz, Wileńska 37, 94-029, Lodz, Poland.
Med Oncol. 2016 May;33(5):51. doi: 10.1007/s12032-016-0763-7. Epub 2016 Apr 18.
In the present study, we analyzed (1) the type of HPV infection and (2) the frequency of loss of heterozygosity and microsatellite imbalance (LOH/MSI) in normal cytology and cervical intraepithelial neoplasia (CIN1-3). The cytological material included: low-grade squamous intraepithelial lesions (CIN1, n = 11), high-grade lesions (CIN2 and CIN3, n = 13), and cytologically normal cells from non-neoplastic cervical samples (n = 8). HPV genotyping was done using RealLine HPV 16/18 kit. We used 20 microsatellite markers from: 1p31.2, 3p14.3, 3p21.3, 3p22.2, 3p24.2, 3p25.3, 7q32.2, 9p21.3, 11p15.5, 12q23.2, and 16q22.1. LOH/MSI was correlated with clinicopathological parameters. The presence of HPV DNA was revealed in 78.13 % samples, including normal cytology. LOH/MSI was the most frequent for: 3p25.3 (39 %), 3p22.2 (20.83 %), 3p24.2 (20 %), and 3p14.3 (16.67 %). It was demonstrated that D3S1234 (FHIT; 3p14.3), D3S1611 (MLH1; 3p22.2), D3S1583 (RARB; 3p24.2), D3S1317 and D3S3611 (VHL; 3p25.3) could differentiate patients with CIN2/CIN3 versus CIN1, showing significantly higher frequency in CIN2/CIN3. LOH/MSI frequency for other than 3p markers was lower, 10-22.2 %. The simultaneous occurrence of LOH/MSI for several markers (OFAL) was higher in CIN2/CIN3. Significant differences in OFAL were found between samples with versus without HPV infection. In HPV-positive patients, significant differences in OFAL were found between normal cytology, CIN1 and CIN2/CIN3. HPV infection influences the increase in LOH/MSI frequency, especially in tumor suppressor gene loci. Several studied microsatellite markers seem to be useful for CIN grading. Hopefully, the obtained results, if confirmed on larger patient cohort, would allow creating a panel of markers supporting clinical diagnosis in patients with HPV infection.
在本研究中，我们分析了：（1）HPV感染类型；（2）正常细胞学及宫颈上皮内瘤变（CIN1 - 3）中杂合性缺失和微卫星不平衡（LOH/MSI）的频率。细胞学材料包括：低级别鳞状上皮内病变（CIN1，n = 11）、高级别病变（CIN2和CIN3，n = 13）以及非肿瘤性宫颈样本的细胞学正常细胞（n = 8）。使用RealLine HPV 16/18试剂盒进行HPV基因分型。我们使用了来自1p31.2、3p14.3、3p21.3、3p22.2、3p24.2、3p25.3、7q32.2、9p21.3、11p15.5、12q23.2和16q22.1的20个微卫星标记。LOH/MSI与临床病理参数相关。在78.13%的样本中检测到HPV DNA，包括正常细胞学样本。LOH/MSI在以下位点最为常见：3p25.3（39%）、3p22.2（20.83%）、3p24.2（20%）和3p14.3（16.67%）。结果表明，D3S1234（FHIT；3p14.3）、D3S1611（MLH1；3p22.2）、D3S1583（RARB；3p24.2）、D3S1317和D3S3611（VHL；3p25.3）可区分CIN2/CIN3与CIN1患者，在CIN2/CIN3中的频率显著更高。除3p标记外的其他标记的LOH/MSI频率较低，为10% - 22.2%。CIN2/CIN3中多个标记同时出现LOH/MSI（OFAL）的情况更高。在有HPV感染和无HPV感染的样本之间发现了OFAL的显著差异。在HPV阳性患者中，正常细胞学、CIN1和CIN2/CIN3之间的OFAL存在显著差异。HPV感染会影响LOH/MSI频率增加，尤其是在肿瘤抑制基因位点。几个研究的微卫星标记似乎对CIN分级有用。有望在更大的患者队列中证实所得结果，从而创建一组支持HPV感染患者临床诊断的标记物。
