Heat-killed probiotic bacteria induce apoptosis of HT-29 human colon adenocarcinoma cell line via the regulation of Bax/Bcl2 and caspases pathway.

Abstract

Apoptosis induction in cancer cells is one of the most efficient ways to treat cancer and find anticancer compounds. The aim of this study was to evaluate the cytotoxic effects of heat-killed indigenous probiotic bacteria and apoptosis induction in the HT-29 human colon adenocarcinoma cell line. The growth-inhibitory effects of probiotic heat-killed Lactobacillus brevis and Lactobacillus paracasei isolated from the traditional Iranian food "Terxine" on the HT-29 cell line were determined by 3-(4,5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide (MTT) assay. Flow cytometry by Annexin-FITC kit, DNA fragmentation assay, 4,6-diamidino-2-phenylindole staining, and the expression of Bax, Bcl2, caspase-3, and caspase-9 were used to analyze apoptosis. MTT results demonstrated that the heat-killed bacteria inhibited the proliferation of HT-29 cells and induced apoptosis in a time-, dose-, and strain-dependent manner. The results demonstrated that both bacteria could induce apoptosis in the HT-29 cell line. Heat-killed probiotic bacteria increased the expression of Bax, caspase-3, and caspase-9 mRNA levels in HT-29 cell lines. Also, heat-killed probiotic bacteria reduced the expression of Bcl2 in HT-29 cells. The heat-killed probiotic bacteria in this study exhibited potent growth inhibitory effects on cancer cells. The results demonstrated that L. brevis has a greater ability to inhibit the growth of HT-29 cells and induce apoptosis, compared with L. paracasei. It is proposed that these bacteria can be used as biological products for the treatment and prevention of cancer, pending further investigation.