| OBJECTIVE:To investigate the effects of cell division cycle associated protein 7(CDCA7;also known as JPO1)expression on the progression and prognosis of human colorectal cancer(CRC).METHODS:At first,the colorectal cancer data was assessed in the TCGA database,and then filter out CDCA7 expressed differentially in colorectal cancer and normal adjacent tissues through differential gene screening,followed by mining the data of CDCA7 on functional and signaling pathways,research heat,reported associated diseases,regulatory relationships,transcription factor prediction and expression in other malignancies in Pubmed,Mesh,KEGG,GO and other databases.The present study aims to verify the role of CDCA7 for human colorectal cancer with human colon cancer tissues and cells:Immunohistochemical was acted to determine protein expression levels of CDCA7 in 104 colorectal cancer tissues and normal colorectal tissues for clinical analysis and survival analysis.15 pairs of colorectal cancer tissues and normal colorectal tissues were detected mRNA expression level of CDCA7 by PCR method.At cellular level,mRNA and protein expression levels of CDCA7 were detected by PCR and Western blot,respectively,using CRC cell lines HT29,RKO,SW620,HCT116 and human normal colonic epithelial cell line NCM460.The relationship between CDCA7 expression level and clinical progression of CRC and survival prognosis in patients with CRC was observed.RESULTS:The bio informatics analysis of CDCA7 and CRC showed:Differential gene screening results showed that the mRNA expression of CDCA7 in CRC tissues was 3.06 times that of normal colorectal tissues,and the difference was statistically significant;gene function and pathway results showed that CDCA7 is significantly associated with colorectal cancer in terms of apoptosis and transcriptional regulation,but there is currently no pathway research about CDCA7.The research hotspot showed that there are currently 22 related literature reports,31 diseases,and 22 upstream targeted miRNAs have been verified;The related disease that have been reported showed that the most relevant report related to CDCA7 is the tumor,but there is currently no research on CDCA7 and colorectal Cancer;Regulatory relationship results show that there are many upstream targeting mRNAs and IncRNAs of CDCA7,such as miR-124-3p,HCP5 and so on,indicating that CDCA7 involves in transcriptional regulation function;Related transcription factor prediction results showed that the most closely related transcription factors with CDCA7 that have been verified are LBP1,ZF5 and IRF1;The expression level of CDCA7 on other malignant tumors showed that expression level of CDCA7 was up-regulated in malignant tumors,such as lymphoma,esophageal cancer,ovarian cancer,leukemia and so on.All the above bioinformatics data showed that there are few experiment studies on CDCA7,and there is no research related to human CRC and CDCA7.Therefore,the present study provides a preliminary study on CDCA7 and CRC to provide a certain experimental theoretical basis for further research in the future.The result of PCR showed that the mRNA expression levels of CDCA7 in CRC tissues were higher than that in adjacent normal colorectal tissues[(1.00±0.00,16.45±2.87,16.00±1.91,0.26±0.07,4.00±0.89,11.96±2.02,9.92±1.65,10.13±0.98,7.78±1.62,3.922±0.85,1.24±0.69,10.85±1.29,5.86±0.93,1.15±0.14,4.79±1.03)vs(18.90±2.19,9.71±1.08,5.31±1.05,2.58±0.24,31.78±3.81,87.43±5.08,1.62±0.29,5.46±1.07,5.94±0.95,9.78±2.04,50.21±5.93,14.42±2.48,48.17±3.96,23.59±2.29,1.33±0.37),P<0.05].The mRNA expression levels of CDCA7 in colorectal cancer cell lines HT29,HCT116,RKO and SW620 were higher than that of normal colonic epithelial cell line NCM460[(0.528±0.10),(1.548±0.02),(1.879±0.15)and(1.936±0.13)vs(1±0.00),all P<0.05].Western blot analysis showed that compared with normal colonic epithelial cells NCM460,the protein expression levels of CDCA7 in CRC cell lines HT29,HCT116,RKO,SW620 were up-regulated[(0.93±0.21),(1.67±0.17),(1.68±0.13)and(1.87±0.55)vs(1.0±0.27),P<0.05].Immunohistochemistry results showed that the positive expression rates of CDCA7 in normal and CRC tissues were 26.92%(28/104)and 75.96%(79/104),respectively,with a statistically significant difference(P<0.05).The above experimental results indicate that expression level of CDCA7 is up-regulated in CRC.The relationship between the expression level of CDCA7 and the clinical characteristic parameters of CRC progression showed that the intensities of CDCA7 immunostaining were significantly correlated to CRC invasion depth(P=0.002),lymph node metastasis(P=0.038),tumor,node,metastasis(TNM)stage(P=0.001)and distant metastasis(P=0.019).However,no significant differences in gender(P=0.533),age(P=0.369),tumor size(P=0.217)and CRC differentiation(P=0.318)were found between high and low CDCA7 expression groups.This indicates that the expression level of CDCA7 is related to the clinical progression of colorectal cancer.Furthermore,patients with low CDCA7 expression exhibited a greater overall survival rate of CRC compared to those with high CDCA7 expression(P=0.012).In conclusion,the results of this study indicate that CDCA7 is highly expressed in human colorectal cancer and has a promoted effect on the clinical progression of CRC and an adverse effect on survival prognosis in patients with CRC.CONCLUSION:CDCA7 is highly expressed in human colorectal cancer and is associated with clinical progression of colorectal cancer.Patients with high expression of CDCA7 have a poor survival prognosis. |
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