Chebulinic acid inhibits MDA‐MB‐231 breast cancer metastasis and promotes cell death through down regulation of SOD1 and induction of autophagy

• Published in: Cell biology international 2020-12, Vol.44 (12), p.2553-2569
• Main Authors: Sharma, Akshay, Mishra, Tripti, Thacker, Gatha, Mishra, Mukul, Narender, Tadigoppula, Trivedi, Arun Kumar
• Format: Article
• Language: English
• Subjects: Actin; Annexin V; Antitumor activity; Apoptosis; Autophagy; Breast cancer; Cadherins; Cancer; Caspase; Cell cycle; Cell death; Cell proliferation; chebulinic acid; Confocal microscopy; Cytotoxicity; Drug resistance; epithelial–mesenchymal transition; Gel electrophoresis; Gelatin; Gelatinase B; Medicinal plants; Mesenchyme; Metastases; Oxidative stress; Phagocytosis; Propidium iodide; SOD1; Superoxide dismutase; Wound healing
• ISSN: 1065-6995; 1095-8355
• DOI: 10.1002/cbin.11463

Triple‐negative breast cancers (TNBC) are highly aggressive and drug resistant accounting for majority of cases with poor outcome. Purified natural compounds display substantial anticancer activity with reduced cytotoxicity providing a new avenue to combat TNBC. Chebulinic acid (CA), a polyphenol derived from the fruits of various medicinal plants has potent anticancer activity. Here, we demonstrate that CA shows significant cytotoxicity against triple negative MDA‐MB‐231 cells. CA exhibited cytotoxicity to MDA‐MB‐231 cells in 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide assay. Further, CA mitigated MDA‐MB‐231 cells viability and proliferation as shown by reduced live cell count, crystal violet staining, colony formation assay, soft agar assay and cell cycle analysis. Wound healing assay and trans‐well migration assay demonstrated that CA significantly inhibited migration of MDA‐MB‐231 cells. Also reduced MMP9 expression was observed in CA‐treated cells by gelatin zymography. CA negatively regulated mesenchymal characteristics of MDA‐MB‐231 cells demonstrated by F‐actin staining and reduced expression of N‐cadherin by confocal microscopy and western blot analysis. Annexin V/propidium iodide (PI) and active caspase‐3 staining showed that CA was able to induce apoptosis in MDA‐MB‐231 cells but did not activate caspase‐3. Two‐dimensional gel electrophoresis based proteomic analysis demonstrated that CA regulated proteins belonging to the oxidative stress pathway, apoptotic pathway and proteins with antiproliferative activity. Western blot analysis analysis revealed that CA negatively regulated superoxide dismutase 1 (SOD1) and enhanced oxidative stress in MDA‐MB‐231 cells. SOD1 in‐gel activity assay also showed reduced SOD1 activity upon CA treatment. Overexpression studies with GFP‐LC3 and tandem tagged RFP‐GFP‐LC‐3 also demonstrated enhanced autophagy upon CA treatment. Chebulinic acid inhibits epithelial–mesenchymal transition, breast cancer metastasis and promotes cell death in triple negative breast cancer cells MDA‐MB231 having mesenchyme cell‐like morphology.