Squamous Cell Carcinoma Antigen As Cervical Squamous Carcinoma Patients With Lymph Node Metastasis And Tiny Rna Serum Marker Research

[Background]Cervical cancer is the second most common cancer in women worldwide, results in300,000female deaths each year. About85%pathological type of cervical cancer cases are squamous cell carcinomas (SCC). Lymph node metastasis(LNM)is the most major prognostic factor of the patients with Stage IB or HA, the five-year survival rate of patients declines dramatically from approximately80-95%in patients without lymph node metastases to approximately50-65%in patients with positive lymph nodes. There is no effective serum marker to predict LNM of cervical SCC.Squamous cell carcinoma antigen (SCC-Ag) is the most current widely used SCC marker. Serum levels of SCC-Ag have been found to correlate with tumor stage, tumor size, and residual tumor after treatment, recurrent or progressive disease, and survival in patients. However, SCC is not organ-specific (for cervix) or malignancy-specific. A normal initial SCC level cannot exclude the presence of lymph node metastases and extra-cervical spread, and hence it is of limited use in treatment planning. A number of studies have examined the utility of elevated pre-treatment SCC-Ag as a marker for the presence of lymph node metastases. In patients with Stage IB or IIA squamous cell cervical cancer, sensitivity of an elevated pre-treatment level of SCC to detect lymph node metastases has ranged from60to87%with specificity ranging from41to91%.[Objective]To establish the overall diagnostic accuracy of the measurements of serum Squamous cell carcinoma antigen (SCC-Ag) for lymph node metastasis (LNM) in squamous cell cervical cancer.[Method]After a systematic review of current studies, sensitivity, specificity, and other measures of the value of serum SCC-Ag in the diagnosis of LNM were pooled by using random effects models. Published studies on evaluation of SCC-Ag to diagnose LNM in cervical cancer were retrieved from The Cochrane Library, Cochrane Central Register of Controlled Trials, Pubmed, Embase, China National Knowledge Infrastructure (CNKI) databases, WanFang Data, and VIP Information in both English and Chinese. Two reviewers independently assessed the methodological quality of each study with the tool of QUAD AS. Statistical analyses were performed by Meta-Disc1.4and Stata11.0software. Meta-analyses of the reported sensitivity and specificity of each study and Summary Receiver Operating Characteristic curve were performed. [Results]Ten studies met the inclusion criteria for the analysis. After testing the heterogeneity of the included studies, a random effect model was selected to calculate the pool weighted index with95%confidence interval:the sensitivity was0.60(95%CI:0.54～0.65), the specificity was0.76(95%CI:0.73～0.78), the DOR was5.38(95%CI:3.27～8.87), and the AUC of SROC was71.26%. The subgroups were analyzed to identify the sources of heterogeneity maybe according to the cutoff.[Conclusion]SCC-Ag has certain diagnosis value in cervical cancer LNM cases, but it is not a perfect index to identify LNM cases from cervical cancer. [Background]Cervical cancer is the second most common cancer in women worldwide, results in300,000female deaths each year. About85%pathological type of cervical cancer cases are squamous cell carcinomas (SCC). Lymph node metastasis (LNM)is the most major prognostic factor of the patients with Stage IB or II A, the five-year survival rate of patients declines dramatically from approximately80-95%in patients without lymph node metastases to approximately50-65%in patients with positive lymph nodes. There is no effective serum marker to predict LNM of cervical SCC.MicroRNAs(miRNAs) are small non-coding RNAs of approximately22nt size, and modulate differentiation, growth, apoptosis and proliferation of cells, regulate gene expression at the post-transcriptional level. Because of their stability and presence in almost all body fluids, microRNAs constitute a novel class of non-invasive biomarkers for disease diagnosis, however, there is no report about serum microRNA in cervical SCC patients. [Objective]Find a few serum microRNAs as potential novel LNM biomarkers in cervical SCC patients, evaluation the LNM predict value of the marker miRNAs in squamous cell cervical cancer.[Method]We used hybridization arrays to compare the microRNA expression profile in cervical SCC samples among patients with LNM or without LNM. In total,1450microRNAs were examined, using P<0.05and expression intensity>1000,89microRNAs were found to fit our inclusion criteria (62up and27down). Using quantitative real-time PCR, we examined expression of these89microRNAs in cervical (cancer) tissue and in serum from80cancer patients and20healthy women. Upon comparing LNM patients (n=40) to non-LNM patients (n=40) and healthy controls, we compared microRNA expression in different groups of patients and used the area under the ROC curve (AUC)>0.70as our inclusion criteria. Using regression analysis, we integrated the marker microRNAs into a comprehensive factor and drew the fitted binormal ROC curves, compare to serum squamous cell carcinoma antigen (SCC-Ag), access the predictive value.[Results]We identified only7microRNAs that were up-regulated in both tissue and serum of cervical SCC patients with LNM. These microRNAs were miR-1246, miR-20a, miR-2392, miR-3147, miR-3162-5p, miR-4484, and miR-4667-5p. Except for miR-4667-5p, the tissue expression of other six microRNAs was consistent with the serum expression. The AUC of the serum comprehensive factor predicting LNM was0.932(sensitiyity0.856, specificity0.850).The prediction value of serum microRNA was inferior to tissue microRNA (AUC0.992, sensitivity0.967, specificity0.950;P=0.018). However, it is by far superior to serum SCC-Ag (AUC0.713, sensitivity0.612, specificity0.700; P<0.0001). Therefore, it is a good predictor of LNM with clinical value.[Conclusion]Serum microRNAs might have great potential to serve as novel, noninvasive biomarkers for LNM in early stage cervical SCC. MiR-1246, miR-20a, miR-2392, miR-3147,miR-3162-5p and miR-4484are potential biomarkers. [Background]MiR-1246was found in human embryonic stem cells in2008, previous studies have shown that miR-1246might be a potential marker of tumor such as esophageal squamous carcinoma. Preliminary study results show that miR-1246in cervical squamous carcinoma patients with lymph node metastasis (LNM), either in cancer tissue or serum specimens, it is up-expression and it may be a oncogene miRNA.[Objective]To investigate the effect of miR-1246on biological activity of cervical squamous cancer cell SiHa. Prove target genes by experiment, Preliminary discussion biological mechanisms of one of miR-1246.[Method]MiR-1246mimics/inhibitor were chemically synthesized and transfected transiently into SiHa cells. The cell proliferation was measured by MTT method. The cell invasiveness and migration was measured by matrigel invasion assay. N-cadherin (CDH2) protein was measured by flow cytometry. Thrombospondin2(THBS2) protein was measured by western blotting. The dual luciferase reporter vectors containing3’UTR of THBS2were constructed by using Dual-Luciferase Reporter Assay System, and then the relative activity of firefly luciferase was detected to confirm the binding site of miR-1246on THBS2.Transfect miR-1246into the cervical cancer cell SiHa, Contrast the proliferation, invasion and migration ability of SiHa cell between before and after transfection; predict target gene of miR-1246by biological software, and verify the gene in both protein and gene level.The transfection of miR-1246can promote SiHa cell proliferation, invasiveness and migration (P<0.05). Biological software predicts that CDH2and THBS2were target genes of miR-1246. After miR-1246transfection, the CDH2protein expression in SiHa cell didn’t change and THBS2protein was down-expression. Cotransfection with miR-1246and Dual-Luciferase plasmid (containing THBS23’UTR) could significantly decrease the relative activity of firefly luciferase. The test of Dual-Luciferase proved that miR-1246can bind the3’UTR of THBS2specifically.[Conclusion]Transfection miR-1246into SiHa cell can promote proliferation, invasiveness and migration, suppressing the expression of THBS2maybe one of the biological mechanism.