ACTIVATION AND PHARMACOLOGICAL MODULATION OF WNT SIGNAL PATH IN HUMAN HCC CELL LINE (HEPG2)

Project no. 23534 · Status: Kapanmış (Yayın bekleniyor) · 2022

Summary

Cancer is a disease which infects cells, who are the basic elements of living organisms. There are dozens of different types of cancer. Hepatocellular carcinoma (HCC) is the most common and most frequent type of liver cancer worldwide. In this study; the activation of the Wnt pathway in the human HCC (HEPG2) cell line and the gene expression stimulated; the expression of the genes encoding the Dishevelled1 (Dvl1) and Frizzled7 (Fzd7) proteins, which are key proteins in the signal transduction pathway after Wnt ligand stimulation, were evaluated. In addition, the pharmacological modulation of salicylic acid and vitamin D molecules, which are cheap and easily accessible chemicals for the expression levels of the Wnt pathway, were also investigated in this cell model.The HCC (HEPG2) cell line has commercially obscured. In line with the work to be done, HCC cells were incubated at 370C in special media until they reached sufficient numbers. Viability analyzes were determined using light microscopy and cell count.Salicylic acid IC50 dose (24. and 48. hours) for HCC (HEPG2) cells was calculated as 45 μM, Dvit-3 IC50 dose (24. and 48. hours) was calculated as 30 μM.In this study, the expression levels of Dvl1 and Fzd7 genes, which directly or indirectly trigger carcinogenesis, were investigated. In this context, RNA expression was performed with the RNA mini kit from the HEPG2 hepatocellular carcinoma cell line. cDNA was obtained from the RNA samples.Taqman ready primers were ordered for the Dvl1 and Fzd7 genes. Mixtures obtained for each gene were loaded into the ABI7000 RT PCR device using Taqman 2x RT-PCR. Housekeeping gene ACTB was used. CT values were taken for comparison of expression values.According to the results obtained, a decrease in the expression of Dvl1 gene was observed in 24-hour Dvit-3 in-vitro treatment. At the same time, it was revealed that V DVL1 gene expression had similar values with the control groups in salicylic acid and salicylic acid + Dvit-3 in-vitro treatment.Although Fzd7 gene expression had similar values to salicylic acid and Dvit-3 in-vitro treatment (p=0.999) at the end of 24 hours, it was significantly lower than the control group. And also, the change in salicylic acid + Dvit-3 in-vitro treatment showed a similar return to other groups.At the end of 48 hours of application, Dvl1 and Fzd7 gene expressions were decreased compared to control group cells.Dvl1 expression values in Dvit-3, salicylic acid and salicylic acid+ Dvit-3 in-vitro treatment did not differ statistically in the experimental groups, but it was observed that they were significantly lower than the control groups.Similarly, after 48 hours of treatment, it was determined that Fzd7 gene expression had a lower mean in the experimental groups as a result of in-vitro treatment.According to the examinations and the results obtained, it was observed that the Dvl1 and also Fzd7 gene expression values decreased in the experimental groups.

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