Cytotoxicity of extracts of Crescentia cujete L. fruit on human cancer cell lines and their effect on the expression of early apoptotic genes and proinflammatory cytokine genes

• Main Author: Jose, Judith Clarisse G.
• Format: text
• Language: English
• Published: Animo Repository 2019
• Subjects: Calabash tree—Analysis; Anti-inflammatory agents; Biochemistry
• Online Access: https://animorepository.dlsu.edu.ph/etd_masteral/6307; https://animorepository.dlsu.edu.ph/cgi/viewcontent.cgi?article=13367&context=etd_masteral
• Institution: De La Salle University

Crescentia cujete L., commonly known as Calabash or Miracle Tree, is widely used in traditional medicine. It has been reported that it possesses several biological activities including anti-cancer and anti-inflammatory activities. This study aimed to investigate the phytochemical components of the crude MeOH extract and to isolate and characterize the trans-cinnamic acid (TCA) from the fruit pulp using GC-MS analysis. It also determined the cytotoxic, genotoxic and anti-inflammatory activities of the plant extracts on human normal and cancer cell lines and the mechanisms of these activities. Results of the cell viability assay using PrestoBlue® resazurin showed that both crude MeOH extract and TCA were cytotoxic against cancer cell lines HCT-116, H69PR and MCF-7 but not to THP-1 and normal HDFn (IC50 ≥100 μg/mL). The crude MeOH extract showed IC50 values of 26.60, 6.662, 3.945 μg/mL for MCF-7, H69PR and HCT-116 cell lines, respectively. The IC50 values generated after treatment of TCA on MCF-7, H69PR and HCT-116 cell lines were 5.108, 5.119 and 4.555 μg/mL, respectively. Treatment exposure to crude MeOH extract and TCA of HCT-116 cells induced upregulation of early apoptotic genes cfos and cjun transcripts. Moreover, genotoxic analysis of HCT-116 cells by COMET assay evidently showed DNA strand breaks or nuclear damage as evaluated by three comet parameters, tail length, %DNA in tail and tail moment. Downregulation of proinflammatory cytokines IL-1β and TNF-α transcripts was observed in lipopolysaccharide (LPS)-treated phorbol myristate acetate (PMA)-induced macrophage after treatment with TCA. However, crude MeOH extract upregulated the expression of both pro-inflammatory genes in LPS-induced macrophages. This study provides evidences that the crude MeOH extract and TCA from C. cujete fruit were cytotoxic to the cancer cell lines but not to the normal cell line tested. This study also provides strong evidence that TCA has an anti-inflammatory effect against LPS-induced macrophages.