Simin Sharifi1, Jaleh Barar2, Mohammad Saeid Hejazi3, Nasser Samadi4. 1. Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran. ; Students' Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran. 2. Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran. ; Research Center for Pharmaceutical Nanotechnology, Tabriz University of Medical Sciences, Tabriz, Iran. 3. Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran. ; Faculty of Advanced Biomedical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran. 4. Research Center for Pharmaceutical Nanotechnology, Tabriz University of Medical Sciences, Tabriz, Iran. ; Faculty of Advanced Biomedical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran. ; Department of Biochemistry and Medical Laboratories, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.
Abstract
PURPOSE: Doxorubicin is administrated as a single agent in first-line therapy of breast cancer to induce apoptosis in tumor cells. Bax, Bcl-xL, Caspase-8 and 9 proteins are involved in induction of apoptosis. The present study describes Bax, Bcl-xL gene expression and Caspase-8 and 9 protein levels in MCF-7 cells incubated with doxorubicin at different doses an incubation times. METHODS: The cytotoxic effects of doxorubicin were studied using MTT assay. MCF-7 cells were treated with three concentrations of doxorubicin (0.1, 0.5, 1 μM) and incubated for 24, 48 and 72 hours then expression levels of Bax and Bcl-xL genes were elucidated by Real-time RT-PCR technique and protein levels of caspase-8 and caspase-9 proteins were measured using ELISA method. Morphological modifications of the cells were also monitored via light microscopic images. RESULTS: Doxorubicin decreased the anti-apoptotic Bcl-xL and increased pro-apoptotic Bax mRNA levels. Doxorubicin induced a significant increase in Bax /Bcl-xL ratio in all doses and incubation times (p<0.05). Highest (more than 10 fold) increase in Bax /Bcl-xL ratio was revealed after 48 h incubation of the cells with in all doses of doxorubicin. Doxorubicin also increased caspase-9 level in a time and dose-dependent manner, while caspase-8 level didn't follow time and dose dependency pattern. CONCLUSION: Our results confirm that doxorubicin induces mitochondrial-dependent apoptosis by down-regulation of Bcl-xL and up- regulation of Bax and caspase-9 expressions.
PURPOSE:Doxorubicin is administrated as a single agent in first-line therapy of breast cancer to induce apoptosis in tumor cells. Bax, Bcl-xL, Caspase-8 and 9 proteins are involved in induction of apoptosis. The present study describes Bax, Bcl-xL gene expression and Caspase-8 and 9 protein levels in MCF-7 cells incubated with doxorubicin at different doses an incubation times. METHODS: The cytotoxic effects of doxorubicin were studied using MTT assay. MCF-7 cells were treated with three concentrations of doxorubicin (0.1, 0.5, 1 μM) and incubated for 24, 48 and 72 hours then expression levels of Bax and Bcl-xL genes were elucidated by Real-time RT-PCR technique and protein levels of caspase-8 and caspase-9 proteins were measured using ELISA method. Morphological modifications of the cells were also monitored via light microscopic images. RESULTS:Doxorubicin decreased the anti-apoptotic Bcl-xL and increased pro-apoptotic Bax mRNA levels. Doxorubicin induced a significant increase in Bax /Bcl-xL ratio in all doses and incubation times (p<0.05). Highest (more than 10 fold) increase in Bax /Bcl-xL ratio was revealed after 48 h incubation of the cells with in all doses of doxorubicin. Doxorubicin also increased caspase-9 level in a time and dose-dependent manner, while caspase-8 level didn't follow time and dose dependency pattern. CONCLUSION: Our results confirm that doxorubicin induces mitochondrial-dependent apoptosis by down-regulation of Bcl-xL and up- regulation of Bax and caspase-9 expressions.
Entities:
Keywords:
Apoptosis; Bax /Bcl-xL; Breast cancer; Caspase-9; Doxorubicin; MCF-7
Authors: J M Zapata; M Krajewska; S Krajewski; R P Huang; S Takayama; H G Wang; E Adamson; J C Reed Journal: Breast Cancer Res Treat Date: 1998-01 Impact factor: 4.872
Authors: H Kojima; K Endo; H Moriyama; Y Tanaka; E S Alnemri; C A Slapak; B Teicher; D Kufe; R Datta Journal: J Biol Chem Date: 1998-07-03 Impact factor: 5.157
Authors: Dolly Patricia Pardo-Mora; Oscar Julián Murillo; Mauricio Rey-Buitrago; Mónica Losada-Barragán; Jaime Fabian Cruz Uribe; Karina Basso Santiago; Bruno José Conti; Eliza de Oliveira Cardoso; Fernanda Lopes Conte; Rafael María Gutiérrez; Orlando Torres García; José Maurício Sforcin Journal: Vet World Date: 2021-04-22
Authors: Z Khodabandeh; I Jamhiri; N Dehghani; H Daneshpazhouh; B Namavar Jahromi; M Dianatpour; S Alaee Journal: Iran J Vet Res Date: 2021 Impact factor: 1.376
Authors: Valerie B Sampson; Nancy S Vetter; Davida F Kamara; Anderson B Collier; Renee C Gresh; E Anders Kolb Journal: PLoS One Date: 2015-11-16 Impact factor: 3.240