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Updated: Jan 9, 2026

Evaluating Cell Death Using Cell-Free Supernatant of Probiotics in Three-Dimensional Spheroid Cultures of Colorectal Cancer Cells
Published on: June 13, 2020
Evaluation of Lactobacillus delbrueckii Supernatant Effects on Cell Cycle Regulation, Apoptosis Induction, and Gene
Mohammad Reza Majid Khoshkholgh Pahlaviani1, Amir Emami2, Seyed Mohammad Mehdi Mahmoodi1
1Department of Microbiology, Kaz.C, Islamic Azad University, Kazerun, Iran.
Abstract:
This study investigated the anti-tumor properties of Lactobacillus delbrueckii, a probiotic bacterium, against HT-29 human colorectal cancer cells. The primary objective was to evaluate the effects of Lactobacillus delbrueckii supernatant on gene expression, cell viability, apoptosis, and cell cycle progression in HT-29 cells. In Gilan Province, Iran, a native strain of Lactobacillus delbrueckii was identified from the supernatant of local yogurt. After treating HT-29 cells with different doses of the supernatant, the MTT test was used to calculate the IC50 value 24 hours later. Annexin V/PI staining and flow cytometry were used to evaluate the induction of apoptosis, and real-time PCR was used to measure changes in gene expression. The results showed that the supernatant of Lactobacillus delbrueckii strongly and dose-dependently reduced the growth of HT-29 cells. In addition to causing apoptosis and stopping the cell cycle in the G0/G1 phase, the therapy also increased the number of Sub-G1 cells, which is a sign of cell death. According to gene expression study, anti-apoptotic genes (AKT, Bcl-2) were downregulated while pro-apoptotic genes (PTEN, p53, and Bax) were upregulated. Interestingly, the natural isolate outperformed a conventional strain in terms of anticancer efficacy. These results demonstrate Lactobacillus delbrueckii's potential as a colorectal cancer treatment.

