Unraveling the Catha edulis Extract Effects on the Cellular and Molecular Signaling in SKOV3 Cells

Abstract

Khat (Catha edulis (Vahl) Endl.) is an evergreen flowering shrub used as a stimulant in many regions worldwide including East Africa, the Arabian Peninsula, Europe, and the United States. Chewing leaves of khat induces excitement and euphoria, which are primarily attributed to two major constituents, cathinone and cathine. Khat also contains other important constituents such as cathedulins. A considerable number of studies reported side effects induced by the khat extracts to both embryos and adults. These include teratogenicity and developmental retardation, oral cancer and ulcers, high blood pressure, and myocardial infarction. So far, little attention has been paid to the effects of khat extracts on the molecular signaling interactions. We aimed in this study to investigate this through evaluating the effects of khat extracts on SKOV3, a human ovarian adenocarcinoma cell line. We show, by in vitro assays, that khat induces several cellular defects including reduced cell size, cell membrane damage, and apoptosis. At high khat extract concentrations, the cell metabolic activity, cell cycle, and cellular proliferation were affected. RT-qPCR analysis showed an increase in the gene expression of the apoptotic marker BAX, the tumor suppressor p53, and the inflammatory cytokine IL-6. Protein expression analysis by immunostaining showed downregulation of β-catenin, E-cadherin, and Ki-67 and upregulation of FZD8 and SPRY2, suggesting that Wnt and FGF signaling were implicated. SwissTargetPrediction in silico analysis showed that khat constituents cathine, cathinone, catheduline K2, and catheduline E5 bind to family A G-protein-coupled receptor, cause many neurological diseases and disorders such as Alzheimer's, schizophrenia, depression, and anxiety, and induce many ovarian cancer-related diseases. The analysis also showed that important signaling pathways such as CREB, Wnt, FGF, IL-6, and ERK/MAPK, and that of the endometrial cancer, and cell cycle were implicated. Upstream regulators of cathine and cathinone were found to potentially target several molecules including interleukin-8, MMP2, PLAU, and micro-RNAs. In conclusion, khat induces significant cellular and molecular changes that could potentially cause a wide range of serious diseases and syndromes. Such an impact could have a heavy burden on the health care system in the countries where khat is consumed.

The Prognostic Value of the Developmental Gene FZD6 in Young Saudi Breast Cancer Patients: A Biomarkers Discovery and Cancer Inducers OncoScreen Approach

“…Ingenuity pathway analysis (IPA) software (Qiagen, United States) ( http://www.ingenuity.com ) has a backend next-generation knowledge base with clarified up-to-date scientific findings from publications, various databases, and related resources ( Abu-Elmagd et al, 2017 ; Jafri et al, 2019 ; Bahlas et al, 2020 ; Abou-Elhamd et al, 2021 ). Here, we used the IPA to perform the core analysis to functionally annotate the genes regulated by FZD6 in BC to identify specific canonical pathways, unique non-directional gene networks, novel molecular signatures, and regulation of cellular, molecular, and bio-functions using the right-tailed Fisher Exact Test and Benjamini Hochberg Correction (BHC) for multiple testing ( p < 0.05) ( Fisher, 1925 ; Benjamini and Hochberg, 1995 ; Benjamini and Yekutieli, 2001 ).…”

Impact of Khat (Catha edulis) Chewing on Inflammatory Markers and Advanced Glycation End Products among Yemeni Women

“…Analysis of the covariants showed that khat chewing increased hs-CRP and interleukin-6 levels. Previous study has reported that khat extracts increase IL-6 gene expression in human ovarian adenocarcinoma cell lines [9]. Interleukin-6 and TNFα expression significantly increased in the cathinone and cathinone-treated mice group [9].…”

Khat, a Cultural Chewing Drug: A Toxicokinetic and Toxicodynamic Summary

“…significantly stimulated testosterone production and had no effect on interstitial viability HL-60, Jurkat, NB4 cell lines, and primary peripheral leukocytes [44] Organic khat extract induced apoptotic cell death, regulated by the activation of cellular caspase −1, −3, and −8 MOLM-13, MOLM-14, NB4 and MV-4-11 cell lines [45] 200 µg/mL Organic khat extract activated a distinct cell death involving mitochondrial damage and morphological features of autophagy SKOV3 [46] 0.01, 0.03, 0.1, 0.3, 1, 3, and 10 mg/mL Khat induces reduced cell size, cell membrane damage, and apoptosis The highest concentrations (1, 3, and 10 mg/mL) affected cell metabolic activity, cell cycle, and cellular proliferation…”

“…In silico [46] Khat constituents (cathine, cathinone, and catheduline): bound to family A of G-protein-coupled receptors and altered several signalling pathways (CREB, Wnt, FGF, IL-6, and ERK/MAPK)…”

“…In silico study suggested that khat constituents (cathine, cathinone, and catheduline) bind to family A of G-protein-coupled receptors. Furthermore, significant players (cAMP-response element binding protein (CREB), fibroblast growth factor (FGF), Interleukin 6 (IL-6), extracellular signal-regulated kinase 1/2 (ERK) and mitogen-activated protein kinase (MAPK)) implicated in endometrial cancer and the cell cycle were altered [46]. Table 2 recaps all the studies cited above.…”