BackgroundTo assess the feasibility of validating microRNA (miRNA) profile related to paclitaxel-sensitivity in formalin-fixed paraffin-embedded (FFPE) samples of serous ovarian carcinoma (OC) patients.MethodsDeregulated miRNAs identified by miRNA microarray were further detected in 45 FFPE OC samples using Realtime PCR. Correlations between paired FFPE and frozen tumor samples were analyzed. Survival times were compared between 6 high and low miRNAs groups. Western blot and luciferase reporter assay were used for validating the target of miRNA.ResultsSixteen up-regulated miRNAs and twenty-three down-regulated miRNAs were revealed in pacilitaxel-resistant ST30 cells. The up-regulated miRNAs (miR-320a, 22 and 129-5p) and down-regulated miRNAs (miR-9, 155 and 640) were confirmed in paclitaxel-resistant FFPE tumor samples, compared with paclitaxel-sensitive samples. Higher miR-9 and miR-640 showed better survival time in OC patients. Expressions of miR-9, 155 and 22 in FFPE samples were closely mimicked by those in frozen tissues. RAB34 was validated as a direct target of miR-9.ConclusionsWe validated miRNA profile in pacilitaxel-resistant OC using FFPE samples, which might enable treatment stratification and help us to predict outcomes in OC patients. FFPE samples are feasible materials for miRNA research.

BackgroundPapanicolaou (Pap) triage, with high specificity, has been recommended for primary Human papillomavirus (HPV) testing but is flawed by poor sensitivity and cytologist dependence. We evaluated the potential role of microRNA (miRNA) detection in cervical exfoliated cells in HPV-positive women from a clinic-based population.MethodsPrimary HPV testing as well as Pap test were performed on all eligible women. Six miRNAs (miR-424/miR-375/miR-34a/miR-218/miR-92a/miR-93) were detected by RT-qPCR in cervical exfoliated cells. All HPV-positive women underwent colposcopy and further biopsy if indicated. Mann–Whitney U test, the receiver operating characteristic curve, logistic regression, and Pearson’s Chi-square were used to assess data. All tests of statistical significance were two-sided.ResultsA total of 1021 eligible HPV-positive women were enrolled. The expression of miR-424/miR-375/miR-34a/miR-218 in high-grade cervical intraepithelial neoplasia (CIN) and abnormal cytology was statistically significantly lower than that in low-grade CIN and normal cytology, respectively (all P < .05). Compared with the Pap test, both miR-424 and miR-375 detection achieved higher sensitivity (76.0% and 74.9% vs 63.8%, P < .05), higher negative predictive value (NPV) (85.7% and 85.4% vs 79.3%, P < .05), and comparable specificity while identifying CIN2 or worse (CIN2+). Similar results were achieved while identifying CIN3+. Multi-marker panels based on miR-424, miR-375, and miR-218 further improved the performance over any single miRNA test or Pap test.ConclusionSingle miR-424 or miR-375 detection and miR-424/miR-375/miR-218–based multimarker panels in cervical exfoliated cells show superior performance over Pap triage for high-grade CIN identification in a clinic-based population. Detection of miRNA may provide a new triage option for HPV-positive women.

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LEEP, carcinoma in situ, menopausal status, and larger area of lesion are risk factors for positive margins. For women with CIN 3 and positive margins, follow-up at an interval of 6 months or repeat excision are treatment options. However, when repeat excision is technically impossible, whether simple hysterectomy or radical surgery is a rational treatment option requires further investigation.

DNA methylation is one of the regulatory pathways that modulate human papillomavirus (HPV) gene expression. To obtain detailed methylation information on crucial areas of the long control region (LCR) of HPV 16 and to clarify the significance of methylation in clinical cervical lesions, 80 clinical samples were examined to determine the methylation status of the HPV 16 promoter and enhancer core using bisulfite modification and pyrosequencing. Seventy samples [26 of cervical carcinoma (CC), 13 of cervical intraepithelia neoplasia (CIN) III, 17 of CIN I-II and 14 of asymptomatic HPV 16 infection] were successfully examined. Analysis of the general methylation status of HPV 16 LCR in the 70 clinical specimens revealed 43 (61.4%) with methylation in the promoter and/ or enhancer core of HPV 16. The proportion of methylated samples was highest in CC specimens (84.6%), followed by asymptomatic infection (71.4%) and CIN III (46.2%), while the proportion of methylated samples was lowest in CIN I-II specimens (29.4%). The methylation status of eight CpGs in HPV 16 LCR was determined in detail. In general, the methylation of CpGs was more common in the promoter than in the enhancer core region. The methylation frequencies of the eight CpGs ranged from 14.6±7.2 to 33.7±23.0% in individual methylated CpG cases. The methylation pattern of all eight CpGs methylated in the promoter and enhancer core was more common in CC, and the pattern of scattered methylated CpGs was relatively more prevalent in asymptomatic infections. Our study demonstrates that DNA methylation is a common phenomenon in HPV 16 LCR clinical specimens, and may function as a host defense mechanism. While hypomethylation is probably associated with the initiation of neoplasia, hyper-methylation in cervical cancer may be a reflection of the host defense mechanism. In the regulation of transcription, methylation is of more importance in the HPV 16 promoter than in the enhancer core.