Effect Of Circ-ARHGEF28 On The Biological Function Of Breast Tumor Cells

Objective Investigate the expression of circ-ARHGEF28 in breast cancer tissues,peripheral blood and cells,evaluate the possibility of its expression for clinical diagnosis of breast tumors,predict miRNAs that bind to circ-ARHGEF28.To investigate the role of circ-ARHGEF28 in the process of breast cancer proliferation and metastasis.Methods Collected a total of 20 pairs of cancer tissue and adjacent tissue samples from patients diagnosed with breast cancer at the General Hospital of Ningxia Medical University from October 2016 to August 2018.Blood samples were collected from 113 patients with breast cancer from August 2017 to September 2018.Fifty blood samples were collected from October 2017 to September 2018 at the Ningxia Medical University General Hospital.The clinical data and pathological examination results of all patients were compared and analyzed.The circRNA chip was used to screen circRNA differentially expressed between breast cancer tissues and adjacent tissues,and 546 high-expressing circRNAs and 1475 low-expressing circRNAs were found(FC?2,P<0.05).Four were verified by qRT-PCR.Candidate circRNA as a Biomarker for Breast Cancer Diagnosis and Treatment.The t-test,curve regression,and ROC curve were used to analyze the relationship between the expression of circ-ARHGEF28 and the clinicopathological characteristics of breast cancer patients.Construct circ-ARHGEF28 overexpression vector and verify its effect on tumor cell proliferation and apoptosis in breast cancer cells;screen,predict and verify the miRNAs potentially binding to circ-ARHGEF28 through the database.qRT-PCR verified the expression of miRNA bound to circ-ARHGEF28,combined with GO analysis and KEGG to analyze the effect of circ-ARHGEF28 on breast cancer cell function.Results Circ-ARHGEF28 is significantly lower expressed in breast cancer tissues compared to adjacent cancer tissues;circ-ARHGEF28 is significantly lower expressed in peripheral blood of breast cancer patients compared to healthy people;circ-ARHGEF28 is significantly lower in breast cancer than normal breast cell lines Significantly low expression in cells.ROC curve analysis showed that the AUC of circ-ARHGEF28 was 0.823,the sensitivity was 82.3%,and the specificity was 68.0%(P <0.01).The expression of circ-ARHGEF28 is closely related to tumor tissue grade,estrogen receptor ER positive,tumor metastasis and TNM stage(P <0.05).The circ-ARHGEF28 overexpression vector was successfully constructed and successfully transfected into breast cancer cells.CCK-8,cell scratches,cell cycle and apoptosis experiments confirmed the effect of this gene on tumor cell proliferation and apoptosis.High-throughput sequencing found 28 over-regulated miRNAs and 5 down-regulated miRNAs after overexpression of circ-ARHGEF28,and target differential gene expression and functional enrichment analysis of these differentially expressed miRNAs.GO analysis of biological pathways of differentially expressed miRNA target genes in the regulation of cell proliferation,cell differentiation,WNT signaling pathway,and apoptosis enrichment,KEGG analysis of target genes in PI3K-Akt signaling pathway,MAPK signaling pathway,TGF-beta signaling The pathway,p53 signaling pathway and Ras signaling pathway are significantly enriched.Screening based on database miRNA target gene prediction,pathway enrichment analysis,and research literature,finally identified 12 miRNA targets with significant differences.After verification by qRT-PCR,the differences were found to be the same as sequencing results.Bioinformatics predicted that circ-ARHGEF28 and miR-375 have binding sites,and further analysis found that miR-375 is mainly related to the regulation of cell proliferation and metastasis.Conclusion The relative expression of circ-ARHGEF28 in the peripheral blood of breast cancer patients has significant specificity,which is closely related to the occurrence of breast cancer and can be used as a new type of biomarker for the diagnosis of breast cancer.circ-ARHGEF28 has a great correlation with lymph node metastasis,histopathological staging,and typing of breast tumors,and may be used as a peripheral blood marker for staging and typing of breast cancer.The target gene of miRNA bound by circ-ARHGEF28 can inhibit the proliferation and differentiation of breast tumor cells and induce apoptosis,and circ-ARHGEF28 can be used as ceRNA to adsorb miR-375,which has the potential to regulate the proliferation and metastasis of breast cancer.