Effect Of Expression Level Of Hsa-circ-0068669 On Proliferation And Metastasis Of Breast Cancer Cells And Its Target Gene Prediction

Objective Breast cancer ranks the first in the incidence of female cancer,and clinical diagnosis mostly relies on imaging and invasive examination,lacking of gene-related detection and treatment methods in clinical application.Currently,studies on non-coding rnas including circular RNA microRNA and miRNA have shown that they play an important regulatory role in the process of tumor development.The purpose of this study was to investigate the role of circrna hsa-circ-0068669 expression level in the proliferation and metastasis of breast cancer cells,and to predict the possibility of finding target genes with targeted regulatory effects.Methods PCR amplification of cDNA and gDNA with divergent primers and convergent primers were used to verify the circular structure of hsa-circ-0068669.QPCR method was used to detect the relative expression of hsa-circ-0068669 in human breast cancer cell lines,as well as the expression difference of hsa-circ-0068669 in breast cancer tissues and adjacent tissues.Using liposome transfection mediated points method from the circ-0068669 expression plasmid,raised from the expression of the circ-0068669,using qPCR method after testing the expression from the circ-0068669 relative to express,through the experiment of determined by MTT method to detect from the circ-0068669 expression in human breast cancer cells MDA-MB-231 and the influence of MCF-7 cell proliferation activity,by using the method of cell scratch test effect on breast cancer cell migration ability,Transwell assay detected that the invasion ability of MDA-MB-231 cells and MCF-7 cells was affected by the overexpression of hsa-circ-0068669.The possible miRNA binding with hsa-circ-0068669 was predicted by bioinformatics method and detected by qPCR,so as to further study the mechanism of hsa-circ-0068669 regulating the biological behavior of cells.Results The convergent primers in both cDNA group and gDNA group had amplification products,while the divergent primers in gDNA group had no amplification products,which proved the circular structure of hsa-circ-0068669.In human breast cancer cell lines MDA-MB-231(0.0041±0.0030,P < 0.05)and MCF-7(0.2262±0.0289,P < 0.05),the expression of hsa-circ-0068669 was lower than that of McF-10 a in non-tumorigenicity cell lines.The expression level in P < 0.001 was significantly lower than that in para-carcinoma tissues(0.8608±0.6698),and the expression level of hsa-circ-0068669 in MDA-MB-231 and MCF-7 cells was up-regulated.The MTT assay showed that the proliferation activity of MDA-MB-231 cells(F=831.604,P < 0.001)and MCF-7 cells(F=236.494,P < 0.001)was decreased,and the migration of MDA-MB-231 cells(0.4707±0.0336)was decreased.P < 0.05)and invasion ability(P < 0.05)were reduced.The targeted miRNA bound to hsa-circ-0068669 was mir-4639-3p predicted by ReRNA,and the expression levels of mir-4639-3p(0.3800±0.0984,P < 0.05)in MDA-MB-231 and MCF-7 cells that overexpressed hsa-circ-0068669 were significantly down-regulated by qPCR.Conclusions The expression of hsa-circ-0068669 is low in human breast cancer cells MDA-MB-231,MCF-7 and breast cancer tissues.Up-regulating the expression level of hsa-circ-0068669 can inhibit the proliferation,cell migration and invasion levels of MDA-MB-231 cells and inhibit the proliferation of MCF-7 cells.Hsa-circ-0068669 maybe regulate the expression level of mir-4639-3p to participate in the development and metastasis of human breast cancer,which may be related to the survival rate and treatment prognosis of breast cancer patients.