Did you know that there is a “Hash, Marihuana & Hemp Museum” located in Amsterdam, Netherlands?
Dedicated to education and scientific studies, this world-famous museum has received more than 2 million visitors since 1985. Despite its controversial uses and long banning histories, cannabis (commonly referred to as marijuana) and its chemical derivatives have been found to have significant medical benefits and may help treat a wide scope of diseases. Today we will briefly talk about the important compound derived from the cannabis and highlight one recent study on its epigenetic function in skin genes.
Cannabinoids and endocannabinoids
Cannabinoids, first discovered in cannabis, refer to the class of compounds derived from cannabis that act on cannabinoid receptors. Cannabinoids are currently used in cancer patients to palliate wasting, emesis and pain that often accompany cancer.
In 1975, Munson, Harris, Friedman, Dewey and Carchman discovered that some cannabinoids (e.g., Δ9-THC, Δ8-THC and cannabinol) inhibited the growth of Lewis lung adenocarcinoma cells in vitro as well as in vivo (1). This finding led to significant advancement in the use of cannabinoids for cancer treatment.
Cannabinoid receptors are a class of G protein-coupled receptors that are spread throughout the body. Together with their endogenous ligands, endocannabinoids and enzymes that regulate their biosynthesis and degradation, they form the endocannabinoid system (ECS).
The endocannabinoid system is involved in a broad range of functions in a growing number of physiopathological conditions. A thorough review on its therapeutic exploitation was published in 2004 (2). Recently, ECS has also been reported to play a role in controlling skin differentiation, growth and apoptosis (3).
Cannabidiol and keratin 10 (KRT10)
Up to date, more than 113 active cannabinoids have been identified in cannabis. Cannabidiol (CBD) is one of the most abundant cannabinoids and is therefore the most recognized and studied. CBD appears to have a wide-spectrum of potential medical applications in epilepsy, breast cancer and schizophrenia, among other conditions.
Pucci et al. recently investigated CBD’s effects on the expression of keratin 10 – or KRT10, a skin differentiation gene – as well as CBD’s impact on the DNA methylation levels of keratin 10 (4). The researchers induced cell differentiation of an adult human skin cell line (HaCaT) and treated them with various cannabinoids, including CBD.
Pucci et al. first performed RT-PCR with the QuantiTect Reverse Transcription Kit. Subsequently, they analyzed the relative abundance of mRNA derived from the cells using the QuantiFast Multiplex PCR Kit. They then tested DNA methylation levels using a DNeasy kit and bisulfite reactions.
Pucci et al. found that CBD can reduce the expression of all selected genes in differentiated adult human skin cell lines by increasing DNA methylation of the KRT10 gene. The study’s discussion further supported that CBD and CBG are potential transcriptional suppressors controlling cell proliferation and differentiation. These findings highlight CBD’s potential for treating skin diseases.
Read more about the researchers’ experiment setup here.
To complete your own quantitative analysis of gene expression, check out the QuantiNova Reverse Transcription Kit that can be used in combination with the kits mentioned above.
Quantification of multiple targets
QIAGEN has recently released a new multiplex PCR kit. Inheriting the strengths of our QuantiNova series, the new QuantiNova Multiplex PCR kit allows sensitive detection of up to 5 targets in 1 tube.
The hot start and PCR buffer system ensures highly sensitive qPCR performance on any real-time cycler without the need for optimization; it also provides automated reaction setup options at room temperature. The built-in tracking system for visual identification of correct pipetting minimizes human errors, and inclusion of the Internal Control RNA helps monitor successful reverse transcription and qPCR.
- 1. Munson, A.E., Harris, L.S., Friedman, M.A., Dewey, W.L. and Carchman, R.A. (1975) Antineoplastic activity of cannabinoids. J Natl Cancer Inst. 55, 597-602. (Link)
- 2. Di Marzo, V., Bifulco, M. and De Petrocellis, L. (2004) The endocannabinoid system and its therapeutic exploitation. Nat Rev Drug Discov. 3, 771–784. (Link)
- 3. Bíró, T., Tóth, B.I., Haskó, G., Paus, R. and Pacher, P. (2009) The endocannabinoid system of the skin in health and disease: novel perspectives and therapeutic opportunities. Trends Pharmacol Sci. 30, 411–420. (Link)
- 4. Pucci, M., Rapino, C., Di Francesco, A., Dainese, E., D’Addario, C. and Maccarrone, M. (2013) Epigenetic control of skin differentiation genes by phytocannabinoids. Br J Pharmacol. 170, 581–91. (Link)