Mutant P53 modulation by cryptolepine through cell cycle arrest and apoptosis in triple negative breast cancer

• Published in: Biomedicine & pharmacotherapy Vol. 179; p. 117351
• Main Authors: Qayoom, Hina, Mir, Manzoor A.
• Format: Journal Article
• Language: English
• Published: France Elsevier Masson SAS 01.10.2024; Elsevier
• Subjects: Apoptosis; Apoptosis - drug effects; Cell cycle arrest; Cell Cycle Checkpoints - drug effects; Cell Line, Tumor; Cell Movement - drug effects; Cell Proliferation - drug effects; Cell Survival - drug effects; Cryptolepine; Female; Humans; Indole Alkaloids - pharmacology; MCF-7 Cells; Migration; Molecular Docking Simulation; Mutant P53; Mutation; Quinolines - pharmacology; TNBC; Triple Negative Breast Neoplasms - drug therapy; Triple Negative Breast Neoplasms - genetics; Triple Negative Breast Neoplasms - pathology; Tumor Suppressor Protein p53 - genetics; Tumor Suppressor Protein p53 - metabolism; Cell cycle arrest; Cryptolepine; Mutant P53; TNBC; Migration; Apoptosis
• ISSN: 0753-3322; 1950-6007; 1950-6007
• DOI: 10.1016/j.biopha.2024.117351

Triple Negative Breast cancer is an aggressive breast cancer subtype. It has a more aggressive clinical course, an earlier age of onset, a larger propensity for metastasis, and worse clinical outcomes as evidenced by a higher risk of recurrence and a shorter survival rate. Currently, the primary options for TNBC treatment are surgery, radiation, and chemotherapy. These treatments however remain ineffective due to recurrence. However, given that p53 mutations have been identified in more than 60–88 % of TNBC, translating p53 into the clinical situation is particularly important in TNBC. In this study, we screened and evaluated the therapeutic potential of cryptolepine (CRP) in TNBC in-vitro models being an anti-malarial drug it could be repurposed as an anti-cancer therapeutic targeting TNBC. Moreover, the cytotoxicity activity of cryptolepine to TNBC cells and a detailed anti-tumor mechanism in mutant P53 has not been reported before. MTT assays were used to examine the cytotoxicity and cell viability activity of Cryptolepine in TNBC, non-TNBC T47D and MCF-7 and non-malignant MCF10A cells. Scratch wound and clonogenic assay was used to evaluate the cryptolepine’s effect on migration and colony forming ability of TNBC cells. Flow cytometry, MMP and DAPI was used to assess cell cycle arrest and cell apoptosis mechanism. The expression of proteins was detected by western blots. The differential expression of RNAs was evaluated by RT-PCR and the interaction between P53 and drug was evaluated computationally using in-silico approach and in-vitro using ChIP assay. In this study, we found that cryptolepine has more preferential cytotoxicity in TNBC than non-TNBC cells. Notably, our studies revealed the mechanism by which cryptolepine induces intrinsic apoptosis and inhibit migration, colony formation ability, induce cell cycle arrest by inducing conformational change in the mutant P53 thereby increasing its DNA binding ability, hence activating its tumor suppressing potential significantly. Our study revealed that CRP significantly reduced the proliferation, migration and colony forming ability of TNBC cells lines. Moreover, it was revealed that CRP induces cell cycle arrest and apoptosis by activating mutant P53 and enhancing its DNA binding ability to induce its tumor suppressing ability. •TNBC is an aggressive breast cancer subtype that lacks the three hormone receptors.•Chemotherapy remains ineffective in TNBC and there is a need to make it targetable.•TP53 “Guardian of the cancer genome” is found mutated in nearly 80 % of the TNBCs.•CRP activated mutant P53 thereby enhancing its tumor suppressing ability in TNBC.•CRP induced its effect in TNBC via; the P53-dependent intrinsic apoptotic pathway.