The Expression And Clinical Significance Of Circ-CCND1 In Human Colorectal Cancer Tissues

Objective:At present,the internal molecular mechanism of colorectal cancer(CRC)is still unclear,especially for its occurrence and development.Our previous studies have found that mi R-193a-3p can affect CRC cells’ growth and invasion in CRC by regulating the target gene PLAU,Researches also show that mi RNA ’s own expression is also regulated by circ RNA.This study aims to explore and determine the upstream regulatory circ RNA of mi R-193a-3p,and systematically study the relationship between circ RNA and the clinicopathological parameters of CRC at the tissue level and then to find new moleculer related to the occurrence and development of CRC,which may enrich its internal mechanism.Methods:1.By using the bioinformatics methods,such as Starbase,and Circular RNA Interactome,we firstly predict the possible upstream circ RNAs of mi R-193a-3p.Then,the top circ RNAs according to the specific Ago CLIP-seq sequencing results were selected and their expression in CRC cell lines were verified by qRT-PCR experiment.Finally,the target circ RNA with the best expression was choosed to perform further study.2.Fifty pairs of CRC tissues and corresponding adjacent tissues,near more than5 cm from the edge of colorectal cancer tissue,were surgically resected and diagnosed by pathology department in Taizhou People’s Hospital from November 2019 to November 2020.QRT-PCR was used to detect the expression of circ-CCND1 in colorectal cancer tissues and adjacent tissues.The correlations of expression of circ-CCND1 with the clinicopathological parameters,such as age,gender,location,tumor size,tissue differentiation,lymph node metastasis,and TNM stages were analyzed in colorectal cancer,while p<0.05 was considered statistically significant.Results:1.A total of 1883 upstream circ RNA,which may have a regulatory effect on mi R-193a-3p,were screened through bioinformatics methods.According to the ranking of Ago CLIP-seq sequencing results,12 circ RNA with relatively high positions were selected.QPCR was performed in colorectal cancer cell lines to verify their expression.And based on the comparative analysis of cycle threshold(CT)value,circ-CCND1,which is stably expressed and well expressed in the five colorectal cancer cell lines,was finally selected as the object for further study.2.According to the qRT-PCR results of 42 pairs of colorectal cancer tissues and their adjacent normal mucosa tissues,the expression of circ-CCND1 in colorectal cancer tissues was significantly higher than those in adjacent normal tissues(P <0.001).3.Combined with the analysis of clinicopathological parameters of patients with CRC,it was found that the expression of circ-CCND1 in colorectal cancer tissues had a significant positive correlation with the clinical stage of TNM and lymph node metastasis.The expression of circ-CCND1 in patients with III-IV stage were significantly higher than those in patients with III+IV stage(P<0.05).Its expression level was higher in the patients with lymph node metastasis than those without(P<0.05).There were no significant differences in circ-CCND1 levels among patients of different ages,genders and with tumor location,size,and degree of tumor differentiation(P > 0.05).Conclusion:1.The expression of circ-CCND1 in colorectal cancer tissues was significantly higher than those in adjacent normal tissues.It is suggested that circ-CCND1 may promote the occurrence and development of colorectal cancer.2.The expression of circ-CCND1 in colorectal cancer tissues has a significant positive correlation with the clinical stage of TNM and lymph node metastasis.It is suggested that circ-CCND1 may have a potential prognostic value.