Studying the effect of CDH۲-i۲ miR-۲ and CDH۲-i۲ miR-۳,on cell migration

Breast cancer is the most common cancer among Iranian women (۲۴.۴% of all cancers). According to studies conducted in ۲۰۰۵-۲۰۰۶, the incidence of breast cancer in Iranian women was ۱۳.۳%. According to the report of the World Health Organization, the incidence of breast cancer has increased by ۲% annually. On the other hand, miRNA plays a key role as one of the most important post-transcriptional regulators of genes.miRNAs usually prevent the expression of some genes by suppressing or destroying mRNAs and thus affect cellular processes such as growth, proliferation, differentiation and cell death. Any change in the function of these molecules can contribute to the development or progression of cancer.Cell Migration occurs through intrinsic mechanisms such as changes in the cytoskeleton, chemical signaling, and cell adhesion proteins. This inherent ability of cells to migrate in embryonic development plays an important role in the formation of tissues and organs. In cancer, this process goes out of control and cancer cells are separated from the primary tumor and metastasize to other parts of the body.CDH۱-i۲-miR-۳, CDH۱-i۲-miR-۲,a microRNA-tagged domain in intron ۲ and ۳ of the E-cadherin gene, are less known miRNAs that are assumed to play an effective role in cell migration and possibly cancer metastasis. In this way, with a series of bioinformatics studiesand scratch assay and real time pcr We tried to investigate the effect of increasing the expression of these microRNAs. with bioinformatics methods , we tried to reach the genes involved in cell migration, among which we can reach VEGF, TGFB, SNAIL, VIMENTIN, then by transfecting the mentioned micro RNAs into HEK ۲۹۳ cells, the expression effect We increased them and investigated the effect of this microRNA on the target genes with real time PCR. Since our microRNAs are tumor suppressors and the effect of increasing their expression in the target genes like VEGF, TGFB, SNAIL, VIMENTIN was measured with the sample of the control gene gapdh and it was found that the increase in the expression of these tumor suppressor microRNAs decreased the expression of genes involved in cell migration. One of the other methods to prove the reduction of migration is the scratch assay, in which HEK۲۹۳T cells were cultured in a ۲۴-well plate and then incubated for ۲۴ hours. After transfection, a direct wound line was made by scraping with a sterile ۲۰ μl pipette tip across the cell monolayer. The cells were washed with PBS and cultured in HDMEM . The movement of cells towards the wound at different time points is taken under the microscope (with ۴۰x magnification), this shows that increasing the expression of the mentioned microRNAs with this method also reduces migration.

کلیدواژه ها:

microRNA ، non-coding RNA ، gene Regulation ، cell migration