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Cytotoxicity Effect of Lactobacillus casei Cell Extract as Indigenous Probiotic Bacterium on Colon Cancer Cell Line (HT29) and Analysis of Bax and Bcl2 Apoptosis Gene Expression
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E Ali Asgari1   , A Mirzaie 2, H Noorbazargan3 , A Bagheri Kashtali4 |
1- Department of Biology, East Tehran Branch, Islamic Azad University, Tehran, Iran
2- Young Researchers Club and the Elite, East Tehran Branch, Islamic Azad University, Tehran, Iran , A.mirzaie@riau.ac.ir
3- Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran,
4- Department of Biology, Roudehen Branch, Islamic Azad University, Roudehen, Iran |
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Abstract: (9824 Views) |
Abstract
Background and aim: Nowadays, the probiotic bacteria such as lactobacilli are known as preventing factors the development of many diseases including cancer. The aim of this study was to investigate the cytotoxic effect of Lactobacillus casei PTCC 1608 cell extract as probiotic bacteria on colon cancer cell line (HT29) and analysis of Bax and Bcl2 apoptosis gene expression.
Methods: In this experimental study, the cell extract of heat killed L. casei was prepared at 0.01, 0.1, 1, 10, 100 and 1000 µg/ml concentration. The cytotoxicity of various cell extracts on HT29 and HEC293 cell lines were evaluated in 24 hours using MTT assay. Moreover, the Bax and Bcl2 apoptosis gene expression level in HT29 cell line was analyzed using Real Time PCR. The apoptotic effects of cell extract was determined using Flow-cytometry technique. Finally, the collected data were statistically analyzed using one-way analysis of variance with the SPSS/18 software.
Results: The results of MTT assay showed that cell extracts of L. casei was able to reduce the survival rate of HT29 cell line to 0.95±0.44, 73.45±0.21, 51.49±0.87, 39.5±0.45 and 19.7±0.55. Also at the same concentrations in HEC293 cell lines resulted in survival decreased rate of 98.67±056, 94.31±0.87, 26/0 ± 1/92.1±0.26, 91/89.91±0.42, 34/8434±0.55, 29/79.29±0.49. In addition to the Real Time PCR results the expression level of Bax and Bcl2 was increased and decreased in HT29 cell line respectively (2.76 ± 0.54 (P<0.05), 0.21 ± 0.43 (P< 0.05)) in 24 hrs. Moreover, the flow cytometry results indicated the 35.62 % apoptosis in HT29 cell line treated with IC50 value.
Conclusion: The results show that the cell extract of L. casei PTCC 1608 could induced the apoptosis in HT29 cell line and it had low toxicity on HEC293 cell line. Therefore, it seems that L. casei has potential uses as probiotic for pharmaceutical applications including prevention and treatment of colon cancer.
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| Keywords: Lactobacillus casei, Probiotic, HT29 cell line, Cytotoxicity, Apoptosis |
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Full-Text [PDF 815 kb]
(5188 Downloads)
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Type of Study: Research |
Subject:
Microbiology
Received: 2017/01/18 | Accepted: 2017/02/25 | Published: 2017/03/15
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