Potential Diagnosis Value Of The Circulating DHX32 Related MicroRNA In Colorectal Cancer

ObjectiveTo explore the DHX32 gene related microRNAs and was used in diagnosing colon cancer,besides,assessing the diagnostic value of colon cancer as Plasma biomarkers.Methods1.In order to obtain DHX32 gene related microRNAs,the NGS-based microRNA profiles were generated in DHX32 shRNA SW480 cell lysate and control shRNA SW480 cell lysate.2.Screened candidate microRNAs which have the significantly differential expression in colon cancer group and the control group with 18 matched pairs of Colon cancer patients and healthy individuals through the pre-test.3.Assessing SYBgreen PCR method by ago-gel electrophoresis,standard curve and the precision experiment.4.The microRNA,which was screened by pre-test,was further detected in a large scale and combined with clinicopathological characteristics and commonly-used plasma tumor markers to analyze.Moreover,the ROC curve was uesd to estimate the value of diagnose of the microRNA in colon cancer.Results1.38 microRNAs associated with DHX32 gene were detected through high throughput sequencing(|log2(Fold Change)|>=1 and P<0.05).2.In the pre-test,miR-373-3p,sifted from 38 microRNAs,has the significantly differential expression in colon cancer group and the control group(P<0.05).3.The amplification efficiencies of plasma miR-373-3p and cel-miR-39 were 100.716% and 97.398%.Moreover,the stability was excellent,intra-and inter-assay CVs were all less than 5%.4.The expression level of plasma miR-373-3p in control group was lower than colon cancer group and colon benign disease group(P<0.001),besides,The expression level of plasma miR-373-3p was significantly different between colon cancer group and colon benign disease group(P<0.001).In colon cancer group,the expression of plasma miR-373-3p in different TNM stages are all higher than control group.However,no significant differences was detected among different TNM stages.5.The expression quantity of plasma miR-373-3p has not an obvious correlation with age,sex,TNM stage,differentiation,vascular infiltration,nerve invasion,lymph gland invasion,metastasis and histological typing.The expression abundence of plasma miR-373-3p was significantly decreased in 20 postoperative samples(P<0.001).6.The sensitivity,specificity,negative predictive value and positive predictive value of plasma miR-373-3p in diagnosing colon cancer were 78.16%?69.41%?75.64% and 72.34% when the cut-off was-5.721.As for early colon cancer,the sensitivity,specificity,NPV and PPV were 86.36%?69.41?75.64% and 72.34% respectively when the cut-off was-5.722.7.The plasma miR-373-3p had an excellent performance for colonic diseases with a sensitivity of 86.04%,a specificity of 69.41%,a PPV of 81.81% and a NPV of 75.64% as the cut-off was-5.721.8.For predicting colon cancer,Combined analysis of plasma miR-373-3p,CEA,CA-125 and CA-199 could yields a higher diagnostic efficiency(AUC=92.60%,sensitivity of 81.61%,specificity of 94.12%,PPV of 93.42%,NPV of 83.33%)when the cut-off was 0.055.Conclusions1.miR-373-3p is a microRNA which related to DHX32 gene.2.Plasma miR-373-3p could be an auxiliary diagnostic marker in colon cancer and Colonic diseases.Moreover,the diagnosis value is higher than the common tumor markers in early colon cancer.3.Combined analysis of plasma miR-373-3p,CEA,CA-125 and CA-199 could yields a higher sensitivity in diagnosis of colonic cancer.4.The expression abundence of plasma miR-373-3p was significantly decreased in postoperative samples.It suggests that plasma miR-373-3p could be a prognostic marker in colorectal cancer postoperative.