Aberrant Expression Of Serum MiR-17-92 Cluster And Its Clinical Significance In Patients With HPV-positive Cervical Cancer

Objective:Cervical cancer is one of the most common human malignancies with a substantial increase in incidence in China.Human papillomavirus(HPV)infection is now recognized as the main cause of cervical cancer and plays a central role in cervical carcinogenesis.Multiple miRNAs acting as oncgenes or tumor suppressors have been reported dysregulated in high-risk HPV-associated diseases.The miR-17-92 cluster,also known as oncomiR-1,encodes six mature miRNAs:miR-17,miR-18a,miR-19a,miR-19b,miR-20a and miR-92a.It functions as an oncgene in HPV-positive cervical cancer cells.Currently no previous studies have reported analyzing the diagnostic value of serum miR-17-92 cluster.In the present study,we aimed to detect the expression levels of miR-17-92 cluster in serum of HPV ositive patients with cervical cancer and evaluate its clinical significance.Methods:1.Serum samples were collected from 50 HPV-positive patients with cervical cancer and 50 HPV-positive women without malignant disease,and RT-qPCR was performed to analyze the expression levels of serum members of miR-17-92 cluster,including miR-17,miR-18a,miR-19a,miR-19b,miR-20a and miR-92a.The differently expressed miRNAs between both groups were then determined using Mann-Whitney U-test.2.We then enlarged the sample size by using 100 HPV-positive cervical cancer specimens,and adding 100 HPV-positive CIN and 100 HPV-positive controls.The diagnostic values of the differently expressed miRNAs were evaluated compared with Thinprep cytologic test(TCT)detection results using receiver operating characteristic(ROC)curves,and the associations between the clinicopathological characteristics and miRNA expressions were also analyzed.3.We used the Kruskal-Wallis test and Kruskal-Wallisnon test to analyze the serum levels of differentially expressed molecules in HPV-positive patients with cervical cancer.Results:1.The expression levels of serum miR-18a and miR-92a in HPV-positive patients with cervical cancer were significantly higher than that in control and CIN patients,and were significantly higher in CIN patients than that in control,respectively(all P<0.05).2.Receiver operating characteristic(ROC)curve analysis revealed that the area under the ROC curve(AUC)of serum miR-18a and miR-92a for the diagnosis of cervical cancer was 0.810(95%CI:0.749-0.862;sensitivity:89%;specificity:65%)and 0.883(95%CI:0.830-0.924;sensitivity:72%;specificity:91%),respectively.A diagnostic miRNA panel based on combined detection of serum miR-18a and miR-92a was developed by logistic regression model.The AUC of this panel for diagnosing CC was 0.936(95%CI:0.891-0.966;sensitivity:83.5%;specificity:87.8%),which was higher than that of single detection of serum miR-18a or miR-92a(all P<0.05).3.The AUC of serum miR-18a and miR-92a for the identification of CIN3+(CIN grage 3 or wores)was 0.792(95%CI:0.737-0.840;sensitivity:87%;specificity:65%)and 0.799(95%CI:0.744-0.847;sensitivity:55.8%;specificity:92%),respectively.The AUC of our serum miRNA panel for identifying CIN3+ was 0.885(95%CI:0.838-0.921;sensitivity:70.2%;specificity:89.8%),which was significantly higher than that of single detection of serum miR-18a or miR-92a and TCT which was 0.645(95%CI:0.582-0.704;sensitivity:51.9%;specificity:77%)(all P<0.05).4.Using the Kruskal-Wallis test and Kruskal-Wallisnon test,we showed that the levels of serum miR-18a were significantly associated with tumor size(P<0.05),the levels of serum miR-92a were significantly associated lymph node metastasis(P<0.05)and FIGO stage(P<0.05).No significant correlations were found between miRNA expressions and age,tumor type and differentiation(all P>0.05).Conclusions:1.MiR-18a and miR-92a,the members of miR-17-92 clusters,are highly expressed in serum from HPV-positive patients with cervical cancer and CIN.Single miR-18a or miR-92a detection and miR-18a/miR-92a-based marker panel in serum show superior performance over TCT for the identification of high-grade CIN and cerviecal cancer,and have the potential to serve as biomarkers for early diagnosis of cervical neoplasia.Detecton of serum miR-18a and miR-92a may provide a new triage option for high-risk HPV-positive women in cervical cancer screening.2.Serum miR-18a and miR-92a are closely associated with the clinical and pathologic features of HPV-positive patients with cervical cancer.The expression of serum miR-18a is significantly associated with tumor size,and serum miR-92a expression is significantly associated with lymph node metastasis and FIGO stage.Both miR-18a and miR-92a may play important roles in the development of cervical cancer.