Identification of Synergistic Interaction Between Cannabis-Derived Compounds for Cytotoxic Activity in Colorectal Cancer Cell Lines and Colon Polyps That Induces Apoptosis-Related Cell Death and Distinct Gene Expression

Abstract

Introduction: Colorectal cancer remains the third most common cancer diagnosis and fourth leading cause of cancer-related mortality worldwide. Purified cannabinoids have been reported to prevent proliferation, metastasis, and induce apoptosis in a variety of cancer cell types. However, the active compounds from Cannabis sativa flowers and their interactions remain elusive.Research Aim: This study was aimed to specify the cytotoxic effect of C. sativa-derived extracts on colon cancer cells and adenomatous polyps by identification of active compound(s) and characterization of their interaction.Materials and Methods: Ethanol extracts of C. sativa were analyzed by high-performance liquid chromatography and gas chromatograph/mass spectrometry and their cytotoxic activity was determined using alamarBlue-based assay (Resazurin) and tetrazolium dye-based assay (XTT) on cancer and normal colon cell lines and on dysplastic adenomatous polyp cells. Annexin V Assay and fluorescence-activated cell sorting (FACS) were used to determine apoptosis and cell cycle, and RNA sequencing was used to determine gene expression.Results: The unheated cannabis extracts (C2F), fraction 7 (F7), and fraction 3 (F3) had cytotoxic activity on colon cancer cells, but reduced activity on normal colon cell lines. Moreover, synergistic interaction was found between F7 and F3 and the latter contains mainly cannabigerolic acid. The F7 and F7+F3 cytotoxic activity involved cell apoptosis and cell cycle arrest in S or G0/G1 phases, respectively. RNA profiling identified 2283 differentially expressed genes in F7+F3 treatment, among them genes related to the Wnt signaling pathway and apoptosis-related genes. Moreover, F7, F3, and F7+F3 treatments induced cell death of polyp cells.Conclusions: C. sativa compounds interact synergistically for cytotoxic activity against colon cancer cells and induce cell cycle arrest, apoptotic cell death, and distinct gene expression. F3, F7, and F7+F3 are also active on adenomatous polyps, suggesting possible future therapeutic value.

The “Entourage Effect”: Terpenes Coupled with Cannabinoids for the Treatment of Mood Disorders and Anxiety Disorders

“…Synergistic interactions may be found between different cannabinoids ( i.e .,“intra-entourage”) [ 105 ] and between cannabinoids and terpenes ( i.e .,“inter-entourage”) [ 106 ]. Thus, cannabis appliance should be optimized to contain mixtures of these C. sativa -derived combined components.…”

Synergistic cytotoxic activity of cannabinoids fromcannabis sativaagainst cutaneous T-cell lymphoma (CTCL)in-vitroandex-vivo

“…The SCBD extract was fractionated and 5 of the fractions that included HPLC-detected compounds were examined for cytotoxic activity ( Figure 1A). Following 48 h of treatment, fractions S4 and S5 were found to have high cytotoxic activity at 40 μg/mL (a relatively high concentration initially selected based on [19] to differentiate active from non-active fractions). Activity of S4 or S5 was ~12-and ~44-fold greater, respectively, than SCBD crude extract at 40 μg/mL and higher than doxorubicin at a concentration of 300 nM ( Figure 1B).…”

“…GC/MS analyses were carried out following [19], using a HP7890 gas chromatograph coupled to a HP6973 mass spectrometer with electron multiplier potential 2 KV, filament current 0.35 mA, electron energy 70 eV, and the spectra were recorded over 40 to 400 m/z. For concentration, 1 mg of each sample was dried under a gentle stream of nitrogen, and then dissolved in 6 mL of methanol prior to introduction to GC/MS.…”

“…Sixty bp single reads were sequenced on one lanes of an Illumina HiSeq. Transcriptome analysis was carried out as described in [19]. Briefly, the raw-reads were subjected to a filtering and cleaning procedure and FASTX Toolkit (http:// hannonlab.cshl.edu/fastx_toolkit/index.html, version 0.0.13.2) was used to trim read-end nucleotides with quality scores < 30, and to remove reads with less than 70% base pairs with a quality score ≤ 30, using the FASTQ Quality Filter.…”

Full-Spectrum Cannabis Extract Microdepots Support Controlled Release of Multiple Phytocannabinoids for Extended Therapeutic Effect

“…1 − 3 More than 500 natural compounds belonging to a broad range of chemical classes have been identified or isolated from Cannabis . 4 , 5 The most abundant are the phytocannabinoids (plant-derived cannabinoids), a family of terpenophenolic compounds uniquely produced by the plant as secondary metabolites. 6 , 7 Gill et al, 8 showed that the activity of Δ 9 - trans -tetrahydrocannabinol (Δ 9 -THC), the primary active ingredient of Cannabis , is influenced by other compounds present in the plant, promoting the notion of phytochemical synergy in Cannabis pharmacology.…”