Study On Effects Of Curcumin On Proliferation,Migration,Apoptosis And MiRNA Expression Of Human Cervical Cancer Hela Cells

Curcumin is a kind of polyphenol. The results show that curcumin has many beneficial effects on the human body, including reducing blood fat, anti tumor, anti atherosclerosis and anti-virus. Curcumin has been listed by the National Cancer Institute (NCI) as the third generation of anti-cancer drugs in clinical research. In this study, we used MTT, cell scratch test, Annexin V-FITC/PI, miRNA chip technology, miRNA transfection, qRT-PCR, RT-PCR and Westren blot respectively to investigate the effects of curcumin on the proliferation, migration, apoptosis and miRNA expression of HeLa cells, and to screen the key miRNA, target gene and target protein, in order to reveal the anti-tumor effects of curcumin to provide valuable experimental data and scientific results. The results are as follows:1 The effects of curcumin on the proliferation, migration and apoptosis of HeLa cells Human cervical cancer HeLa cells (Hela) as the main research object, setting 8 concentrations of curcumin (0.0,2.5,5.0,10.0,20.0,40.0,60.0,80.0μmol/L), cells were treated for 24,48,72 h respectively, than, the effects of curcumin on the proliferation, migration and apoptosis of HeLa cells are detected by MTT, wound scratch assay and Annexin V-FITC/PI respectively, and screening the best concentration and time for the next test. The results showed that group 48 h,10.0 μmol/L has obvious promoting effect on the apoptosis of HeLa cells; early and late apoptotic cells also occupy a certain proportion in this group. With the increase of the concentration of curcumin, the wound healing rate gradually decreased. So we chose 10.0μmol/L,48 h for the subsequent treatment of curcumin conditions.2 Study on the expression level of miRNA in HeLa cells by curcumingroup 10.0 μmol/L,48 h as the miRNA chip processing condition, screening out the abnormal expression of miRNA, a total of 12 miRNA were found to be up-regulated,10 miRNA down-regulated. Among them, by bioinformatics analysis and literature search selected target genes of hsa-miR-486-3p:protein kinase B beta (protein kinase B beta, AKT2) and epidermal growth factor receptor (epidermal growth factor receptor, EGFR); RT-PCR and Western blot results showed that under the effects of curcumin, AKT2 gene expression had a downward trend, and protein expression was up-regulated, and mRNA and protein expression of EGFR were all up-regulated, they all showed a concentration dependent relationship.3 Transfection and verification of hsa-miR-486-3p in HeLa cellsTransfection of hsa-miR-486-3p inhibitor and mimic in HeLa cells, the optimal dose was screened out by MTT colorimetric assay for cell scratch test, Annexin V-FITC/PI, Western Blot and RT-PCR. The results showed that the high expression of hsa-miR-486-3p could promote the proliferation of HeLa cells, but had no significant effect on cell migration and apoptosis; After transfection of hsa-miR-486-3p inhibitor, compared with the control group, the expression of AKT2 gene was decreased, the expression of protein was increased by 15.4%, the expression of EGFR gene increased, and the protein expression increased by 27.4%, After transfection of hsa-miR-486-3p, AKT2 gene expression increased, the protein expression level decreased by 35.1%, EGFR gene expression decreased, the protein expression of the relative control group decreased by 12.0%. The results suggested that curcumin could down regulate the expression of hsa-miR-486-3p in HeLa cells and affect the expression of the target gene and target protein, which could inhibit the proliferation of HeLa cells.