trans-Anethole Abrogates Cell Proliferation and Induces Apoptosis through the Mitochondrial-Mediated Pathway in Human Osteosarcoma Cells

Abstract

trans-Anethole, the major bioactive component of Illicium verum Hook. commonly known as star anise exhibits various pharmacological activities including anti-inflammatory, antimicrobial, insecticidal, and antitumor. Osteosarcoma is an extremely aggressive malignant bone tumor that affects children and young adults and accounts for around 60% of all sarcomas. The study was planned to evaluate the potential of trans-Anethole against Human osteosarcoma cell line MG-63. The antiproliferative activity of trans-Anethole was assessed by MTT assay. trans-Anethole exhibited apoptotic cell death as monitored by confocal/electron microscopy and flow cytometry studies. Modulation of gene expression was studied by Western blot and RT-PCR analysis. The present study revealed that trans-Anethole inhibited osteosarcoma proliferation in a dose-dependent manner with a GI₅₀ value of 60.25 µM and showed pro-apoptotic activity as analyzed by Annexin V-FITC/PI assay. Flow cytometric analysis revealed that trans-Anethole induced cell cycle arrest at the G₀/G₁ phase with the generation of reactive oxygen species and reduction in mitochondrial membrane potential (ΔΨm). Immunoblotting results showed the increased expression of caspase-9/-3, p53, and decreased expression of Bcl-xL suggesting the involvement of the p53 and mitochondrial intrinsic pathway. This work provides a rationale that trans-Anethole might be considered as a promising chemotherapeutic/nutraceutical agent for the management of osteosarcoma.

Highlights

• trans-Anethole inhibited cell growth and caused G₀/G₁ arrest in Human osteosarcoma MG-63 cell line.

• trans-Anethole led to the loss of mitochondrial membrane permeability along with ROS generation.

• trans-Anethole upregulates the expression of p53, Caspase-9/-3, and downregulate Bcl-xL expression.

Acknowledgments

The authors are thankful to Centre of Emerging Life Sciences and the Department of Botanical and Environmental Sciences, Guru Nanak Dev University, Amritsar, Punjab, India, for providing instrumentation and necessary laboratory facilities to carry out this work.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Additional information

Funding

The authors are thankful to UGC (BSR Program) for providing fellowship to KP, DST-FIST (Grant No. SR/FST/LSI-691/2016(C)) program Government of India, for providing financial assistance.