Expression Of Hsa＿circ＿0002320 In Colorectal Cancer And Its Potential Clinical Research Value

Part One The expression of hsa＿circ＿0002320 in colorectal cancer and its relationship with clinicopathological characteristicsObjective: Circ RNA plays an important regulatory role in a variety of malignant tumor diseases,but the mechanism and clinical value of hsa＿circ＿0002320 in CRC are still unclear.The purpose of this study was to investigate the expression of hsa＿circ＿0002320 in clinical CRC and its relationship with clinicopathological characteristics.Methods: 1.The obtained clinical colorectal cancer(CRC)tissues and adjacent normal colorectal mucosa tissues were fixed with 4% paraformaldehyde,dehydrated,paraffin embedded and sected,and then hematoxylin-eosin(HE)staining was used to observe the pathological conditions.2.Real-time fluorescence quantitative polymerase chain reaction(RT-q PCR)was used to detect the expression of hsa＿circ＿0002320 in 50 cases of CRC tissues and their adjacent normal colorectal mucosal tissues,as well as the preoperative plasma of the 50 cases of CRC patients and the plasma of 100 cases of healthy people.In addition,the expression of hsa＿circ＿0002320 in CRC cell lines(HT29,HCT116)and normal colorectal mucosal cell lines(FHC)was detected.3.To analyze the relationship between the expression of hsa＿circ＿0002320 in CRC tissues and plasma and clinicopathological characteristics.Results: Compared with the adjacent normal colorectal mucosal tissues,the expression of hsa＿circ＿0002320 in CRC tissues was significantly down-regulated(P<0.05).Similarly,the plasma hsa＿circ＿0002320 expression in CRC patients was significantly down-regulated(P<0.05).The expression of hsa＿circ＿0002320 in CRC cell lines(HT29,HCT116)was significantly lower than that in normal colorectal mucosa cells(FHC)(P<0.01).The expression of hsa＿circ＿0002320 in CRC tissues was significantly correlated with the degree of tumor differentiation(P<0.05),and the expression of hsa＿circ＿0002320 in plasma of CRC patients was significantly correlated with the level of CEA(P<0.05).Conclusions: Hsa＿circ＿0002320 was significantly down-regulated in CRC tissues,cells and plasma,which may be used as a biomarker and therapeutic target for diagnosis and treatment of CRC.Part Two Effects of hsa＿circ＿0002320 overexpression on invasion,migration,proliferation and epithelial-mesenchymal transfor-mation of colorectal cancerObjective: To investigate the relationship between hsa＿circ＿0002320 and the invasion,migration,proliferation and epithelial-mesenchymal transform-ation of CRC.Methods: 1.Recombination plasmid hsa＿circ＿0002320 was constructed and overexpressed in CRC cells.2.Real-time quantitative PCR was used to detect the expression of hsa＿circ＿0002320 in CRC cells.3.The effects of overexpression of hsa＿circ＿0002320 on the invasion,migration and proliferation of CRC cells were detected by cell function assay.4.The expressions of E-cadherin,N-cadherin and Vimentin associated with epithelial-mesoplasmic transformation in CRC cells were detected by Western blot.Results: The invasion,migration and proliferation of CRC cells in the overexpressed hsa＿circ＿0002320 group were significantly decreased compared with the negative control group.The expression of N-cadherin and Vimentin in EMT-related proteins was significantly decreased,while the expression of E-cadherin was significantly increased compared with the negative control group(P<0.01).Conclusions: Overexpression of hsa＿circ＿0002320 can inhibit the invasion,migration,proliferation and epithelial-mesenchymal transformation of colorectal cancer.Part Three The role of hsa＿circ＿0002320 in the diagnosis and prognosis of colorectal cancer patientsObjective: To investigate the diagnostic and prognostic value of hsa＿circ＿0002320 in CRC.Methods: 1.The relationship between the expression of hsa＿circ＿0002320,clinicopathological characteristics and overall survival(OS)in CRC patients was analyzed.Receiver operating characteristic curve(ROC)was used to analyze the sensitivity and specificity of hsa＿circ＿0002320 for the diagnosis of CRC.2.Bioinformatics was used to verify the effect of hsa＿circ＿0002320 on the prognosis of CRC patients.Results: The expression level of hsa＿circ＿0002320 was significantly correlated with the OS of CRC patients(P<0.01),that is,the higher the expression of hsa＿circ＿0002320,the longer the OS of CRC patients.The high expression of hsa＿circ＿0002320 had a hazardratio(HR)of 0.161 [95% confidence interval(CI),0.066-0.393;P<0.000].The area under the ROC curve(AUC)of hsa＿circ＿0002320 for CRC diagnosis was 0.823,higher than that of CEA(AUC= 0.764).Kaplan-Meier analysis showed that CRC patients with low expression of hsa＿circ＿0002320 had lower OS than CRC patients with high expression of hsa＿circ＿0002320.Conclusions: Hsa＿circ＿0002320 may be a novel,non-invasive blood biomarker for the diagnosis and prognosis of CRC patients.