Evaluation of genes expression changes in the mitochondrial pathway of apoptosis and cell cycle in AsPC-1 cells treated with Fluorouracil

Khorasani, Negar; Baharara, Javad; Nejad Shahrokhabadi, Khadijeh

Volume 10, Issue 2 (9-2023) nbr 2023, 10(2): 99-107 | Back to browse issues page

‎ 20.1001.1.24236330.1402.10.2.1.6

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Khorasani N, Baharara J, Nejad Shahrokhabadi K. Evaluation of genes expression changes in the mitochondrial pathway of apoptosis and cell cycle in AsPC-1 cells treated with Fluorouracil. nbr 2023; 10 (2) :99-107
URL: http://nbr.khu.ac.ir/article-1-3573-en.html

Evaluation of genes expression changes in the mitochondrial pathway of apoptosis and cell cycle in AsPC-1 cells treated with Fluorouracil

Negar Khorasani

, Javad Baharara ^*

, Khadijeh Nejad Shahrokhabadi

Department of Biology & Research Center for Animal Development Applied Biology, Mashhad Branch, Islamic Azad University, Mashhad, Iran , baharara@mshdiau.ac.ir

Abstract: (2733 Views)

Pancreatic cancer is one of the most deadly and aggressive cancers; Fluorouracil induces apoptosis and cell cycle arrest in cancer cells. In the present study; the effect of Fluorouracil on different stages of the cell cycle and the expression of genes involved in the internal pathway of apoptosis in the AsPC-1 cell line (human pancreatic cancer) were investigated. In order to do so, MTT assay was used to evaluate the cytotoxic effect of Fluorouracil on AsPC-1 cell proliferation; The type of induced cell death and cell cycle changes were investigated by flow cytometry; changes in the expression level of genes (BAX, Bcl-2, APAF-1, Caspase-3, Caspase-9, p53, p21) were examined by Real-time PCR. Quantitative data were analyzed at the significant level of (p<0.05). The MTT assay results showed that Fluorouracil decreased AsPC-1 cell proliferation in a concentration-dependent manner. The results of flow cytometry analysis showed that increased percentage of apoptotic cells in the treated cells; Fluorouracil induces S phase cell cycle arrest in AsPC-1 cells and reduced distribution in the G1 phase. The Real-time PCR results in treated cells showed an increase in the expression of genes in the mitochondrial apoptotic pathway as well as genes effective in regulating the cell cycle. Fluorouracil reduces cell proliferation and induces apoptosis by increasing the expression of genes involved in the Intrinsic apoptotic pathway in AsPC-1 cells; Fluorouracil also caused cell cycle arrest in these cells by regulating the (p53, p21) genes.

Keywords: cell proliferation, flow cytometry, pancreatic cancer, real-time PCR

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