Apoptosis Induced by Ziziphora tenuior Essential Oil in Human Colorectal Cancer Cells

Abstract

Ziziphora (Cacotti in Persian) belongs to the Lamiaceae family (mint group) and is vastly found in Iran and Asia. This traditional medicinal plant is normally used as analgesic and for treatment of particular gastrointestinal diseases. Since colorectal cancer is one of the most common causes of death in the world and the second leading cause of cancer death among adults, there is a pressing need to inhibit this malignancy by using methods with minimal side effects. One of these methods is the use of natural resources such as medical plants. This study is aimed at investigating the expression of apoptosis-related genes in the adjacent culture of colorectal cancer epithelial cells (HT-29) with Ziziphora essential oil (ZEO). The essential oil was extracted from Ziziphora leaves, and its compounds were determined and then added to the HT-29 culture medium at different concentrations. After 24 hours, the HT-29 cells were harvested from the medium and cytotoxicity was analyzed by MTT assay. After MTT assay and determination of the percentage of apoptosis by flow cytometry, RNA extraction was performed and the expression levels of Bax, Bcl-2, caspase 3 (C3), and caspase 9 (C9) were analyzed using newly designed primers by reverse transcription (RT) qPCR method and GeniX6 software. Also, specific antibodies were used for western blot analyses of those molecules. GC analysis revealed 42 different compounds in the ZEO, including pulegone (26.65%), menthone (5.74%), thymol (5.51%), and menthol (1.02%). MTT assay showed that the concentration of 200 μg/ml of ZEO had the highest HT-29 cell death during 24 hours. After incubation with the concentration of 50 μg/ml of ZEO for 24 and 48 hours, caspase 3 and 9 gene expressions in the treated group increased compared to those in the control group ( P < 0.001 ), while the Bcl-2 expression decreased. The results showed that having anticancer compounds, ZEO can increase C3 and C9 and decrease Bcl-2 expressions, causing apoptosis in HT-29 cells in vitro. This can lead to the use of ZEO as a factor for colorectal cancer treatment.

The Effect of Thymus vulgaris on Hepatic Enzymes Activity and Apoptosis-Related Gene Expression in Streptozotocin-Induced Diabetic Rats

“…After that, the membrane was washed three times with TBST and the secondary antibody (1 : 1000) was added to be incubated for 1 h and washed with TBST. Then, band intensities were detected using the chemiluminescent substrate Supersignal Femto kit and band densities were analyzed using the ImageJ 1.52a program (Bethesda, Maryland, USA) [ 27 ].…”

Essential Oils from Côa Valley Lamiaceae Species: Cytotoxicity and Antiproliferative Effect on Glioblastoma Cells

“…As an example, Duymus et al [ 69 ] reported the EO from Vitex agnus-castus L. fruit, enriched in 1,8-cineole, showed high apoptotic activity against both A549 (human lung adenocarcinoma cells) and MCF-7 cells [ 69 ]. Another work stated that treatment with Ziziphora tenuior L. EO (with a high content in pulegone) on colorectal cancer epithelial cells (HT-29) revealed a time-dependent apoptosis mechanism [ 70 ]. It is worth mentioning that the reason for these apoptotic events in these studies seems to be due to the rich content in camphor and pulegone from L. pedunculata and M. cervina , respectively.…”

Species diversity and phylogenetic relationships within the tribe Mentheae (Lamiaceae) in Uzbekistan using ITS sequence data

“…According to Harley et al, [14] Pamir Most of them are source of biologically active compounds and used as antioxidant, antiseptic, immunomodulatory, antimicrobial in traditional medicine and pharmaceutics. Thus, Ziziphora [15][16][17], Salvia [18], Hymenocrater [19], Dracocephalum [20] and other represents of Lamiaceae characterized by a high content of polysaccharides, polyphenols, alkaloids and other carbohydrates [21][22][23].…”

“…8 taxa of Ziziphora are named for the territory of Uzbekistan. Its species are of prime importance in different fields of pharmaceutical, chemical, medicinal, traditional and folk medicines [15][16][17].…”