The Anti-inflammatory and apoptotic effects of L. Officinal extracts on HT 29 and Caco-2 human colorectal carcinoma cell lines

Authors

  • Marzieh Lotfian Sargazi Physiology Research Center, Institute of Neuropharmacology, Kerman University of Medical Sciences, Kerman, Iran
  • Zahra Miri Karam Physiology Research Center, Institute of Basic and Clinical Physiology Sciences, Kerman University of Medical Sciences, Kerman, Iran
  • Ali Shahraki Department of Biology, Faculty of science, University of Sistan and Baluchestan, Zahedan, Iran
  • Mahboobeh Raeiszadeh Herbal and Traditional Medicines Research Center, Kerman University of Medical Sciences, kerman, Iran
  • Mohammad Javad Rezazadeh Khabaz Department of Medical Genetics, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran
  • Abolfazl Yari Cellular and Molecular Research Center, Birjand University of Medical Sciences, Birjand, Iran

Abstract

Background: Colorectal cancer is one of the deadliest malignancies worldwide. Due to the occurrence of side effects related to current standard therapy, researchers are seeking better alternative treatments. For many years, herbs have been a promising source for discovering therapeutic compounds. Therefore, the aim of this study was to investigate the apoptotic and anti-inflammatory effects of Levisticum officinale Koch on HT-29 and Caco-2 colon cancer cell lines.

 Methods and Materials: The ethanol extract of this plant and other different extracts (dichloromethane, petroleum, and residues) were prepared using the maceration method on HT-29 and Caco-2 colon cancer cell lines. The cytotoxic effects were evaluated using the MTT assay, and the half-maximal inhibitory concentration (IC50) values were measured. Additionally, this study analyzed the expression levels of several inflammatory genes (IKKb, IKKa, and REIB) using reverse transcription polymerase chain reaction and western blot. The western blot also assessed the proteins involved in apoptosis, including Caspase, BAX, and Bcl-2 proteins.

Results: Among the extracts, the dichloromethane extract of Levisticum officinale (DELO) had the highest cytotoxic effect, with IC50 values of 106.0±2 µg/mL and 175.3±4 µg/mL in Caco-2 and HT-29 cells after 72 hours, respectively. None of the L. officinale extracts showed a significant cytotoxic effect on non-cancerous cells (3T3 cell line). Furthermore, compared to the control group, the group treated with DELO showed a lower level of expression of inflammatory genes as well as COX-2 protein.

Conclusion: DELO was the most hazardous to both HT-29 and Caco-2 cells among other different extracts (EELO, RELO, PELO). Our expression analysis indicated that the DELO decreased IKKb, IKKa, and REIB gene expression in both HT-29 and Caco-2 cell lines. Additionally, COX-2 protein in the HT29 and Caco-2 cell lines has dramatically lowered, according to Western blot results.

References


Published

2024-07-02

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Section

Original Article