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Bryophyllum pinnatum Induces p53-Dependent Apoptosis of Colorectal Cancer Cells via Increased Intracellular ROS and
Sumoyee Mukherjee1, Sheuli Kangsa Banik2, Sourio Chakraborty3
1Department of Biochemistry and Medical Biotechnology, Calcutta School of Tropical, Medicine, Kolkata, India.
Abstract:
Chemotherapy, radiotherapy and surgical treatments of cancer having several limitations and toxic side-effects, have led researchers to focus towards development of alternative natural plant-based therapeutics that can reduce disease severity. The present research work is mainly focussed towards identifying molecular mechanisms of apoptosis of colorectal cancer cells (HCT116) by perennial herb Bryophyllum pinnatum leaf-extract via both in vitro experimentations and in silico analysis. B. pinnatum leaf extract induced highest cytotoxicity at lowest dose (IC50:0.01 mg/mL) against HCT116 cells with 49.5% (p < 0.0001) cellular death, in comparison to other cancer cell lines. It has arrested HCT116 cell populations at G2/M cell-cycle phase and led to 10 folds (p < 0.0001) and 5.5 folds (p < 0.0001) increased intracellular ROS production in treated groups. ROS production might have led to significant 34.23% and 21.03% (p < 0.0001) apoptosis in treated cells, proved in vitro and in silico, with significant upregulation of p53 (p < 0.0001), BAX (p = 0.0252), CASPASE3 (p < 0.0001) and downregulation of BCL2 (p = 0.0058), leading to increased nuclear p53 (p = 0.0002) accumulation in treated cells, suggesting that the leaf-extract might have induced p53-dependent apoptosis of colorectal cancer cells. The phyto-extract also possess significant gene-modulatory potential as evident from qRT-PCR analysis of oncogenes and tumor suppressor genes. Leaf's bioactive phyto-constituents were elucidated by GC-MS and HPLC-ESI/MS analysis. In silico STITCH analysis provided significant network interactions between these bioactive phyto-compounds and studied proteins. Further Molecular Docking studies revealed strong binding between such docked complexes. Also, predicted major bioactive phyto-constituents of B. pinnatum leaf-extract such as Quercetin, Morin and β-Sitosterol have induced significant (p < 0.0001) apoptosis and increased intracellular ROS, validating their in silico interactions with studied proteins of HCT116 cells. All these studies together demonstrated ability of B. pinnatum to be used as a suitable natural phyto-therapeutic agent for development of chemo-preventive medications against colorectal cancer.
Insights
Bryophyllum pinnatum leaf extract effectively induces apoptosis in colorectal cancer cells by increasing reactive oxygen species and upregulating key apoptosis-related genes. This natural compound shows potential as a chemo-preventive agent against colorectal cancer.
Area of Science:
- Phytomedicine
- Cancer Biology
- Molecular Biology
Background:
- Conventional cancer treatments like chemotherapy and radiotherapy have limitations and toxic side effects.
- There is a growing need for alternative, natural therapeutics, particularly plant-based options, to manage cancer severity.
- Bryophyllum pinnatum is a perennial herb with potential therapeutic properties.
Purpose of the Study:
- To investigate the molecular mechanisms of apoptosis induction in colorectal cancer cells (HCT116) using Bryophyllum pinnatum leaf extract.
- To evaluate the efficacy of B. pinnatum leaf extract through in vitro experiments and in silico analysis.
- To identify potential bioactive compounds and their interactions within colorectal cancer cells.
Main Methods:
- In vitro cytotoxicity assays and cell cycle analysis on HCT116 cells.
- Measurement of intracellular reactive oxygen species (ROS) production.
- Gene expression analysis (qRT-PCR) of apoptosis-related genes (p53, BAX, CASPASE3, BCL2) and oncogenes/tumor suppressor genes.
- Phytochemical analysis using GC-MS and HPLC-ESI/MS.
- In silico analysis including STITCH and molecular docking.
Main Results:
- B. pinnatum leaf extract exhibited significant cytotoxicity against HCT116 cells (IC50: 0.01 mg/mL), inducing 49.5% cell death.
- The extract arrested HCT116 cells at the G2/M phase and significantly increased intracellular ROS production.
- Significant apoptosis (34.23% in vitro, 21.03% in silico) was observed, mediated by the p53-dependent pathway with altered expression of p53, BAX, CASPASE3, and BCL2.
- Bioactive compounds like Quercetin, Morin, and β-Sitosterol were identified and shown to induce apoptosis and ROS production, with validated in silico interactions.
Conclusions:
- Bryophyllum pinnatum leaf extract effectively induces p53-dependent apoptosis in colorectal cancer cells.
- The extract's mechanism involves increased ROS production and modulation of key apoptosis-related genes.
- Identified bioactive compounds like Quercetin, Morin, and β-Sitosterol contribute to the observed anti-cancer effects.
- B. pinnatum demonstrates potential as a natural therapeutic agent for colorectal cancer prevention and treatment.
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