Human laryngeal squamous cell carcinoma cell line release of endogenous anandamide and 2-arachidonoylglycerol, and their antiproliferative effect via exogenous supplementation: an in vitro study
Authors: Petek Korkusuz
Şefik Hoşal
Nilda Süslü
Canset Aydın
Övsen Önay
Sevil Köse
Emirhan Nemutlu
Journal: Cell and Tissue Banking (2021)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: 2-Arachidonoylglycerol; Anandamide; Endocannabinoid; Laryngeal carcinoma.
Affiliations: Department of ENT, Faculty of Medicine, Atilim University, 06830, Golbasi, Ankara, Turkey. ., Department of Medical Biology and Genetics, Faculty of Medicine, Atilim University, Ankara, Turkey., Department of ENT, Faculty of Medicine, Hacettepe University, Ankara, Turkey., Department of Histology and Embryology, Faculty of Medicine, Hacettepe University, Ankara, Turkey., Department of Analytical Chemistry, Faculty of Pharmacy, Hacettepe University, Ankara, Turkey., ENT Clinic, Medicana International Hospital, Ankara, Turkey., Department of ENT, Faculty of Medicine, Atilim University, 06830, Golbasi, Ankara, Turkey.

Abstract:  The level of the major endocannabinoids anandamide (AEA) and 2-arachidonoylglycerol (2-AG) are altered in several types of carcinomas, and are known to regulate tumor growth. Thusly, this study hypothesized that the HEp-2 human laryngeal squamous cell carcinoma (LSCC) cell line releases AEA and 2-AG, and aimed to determine if their exogenous supplementation has an anti-proliferative effect in vitro. In this in vitro observational study a commercial human LSCC cell line (HEp-2) was used to test for endogenous AEA and 2-AG release via liquid chromatography-tandem mass spectrometry (LC-MS/MS). The anti-proliferative effect of AEA and 2-AG supplementation was evaluated via WST-1 proliferation assay. It was observed that the HEp-2 LSCC cell line released AEA and 2-AG; the median quantity of AEA released was 15.69 ng mL-1 (range: 14.55-15.95 ng mL-1) and the median quantity of 2-AG released was 2.72 ng -1 (range: 2.67-2.74 ng mL-1). Additionally, both AEA and 2-AG exhibited an anti-proliferative effect. The anti-proliferative effect of 2-AG was stronger than that of AEA. These findings suggest that AEA might function via a CB1 receptor-independent pathway and that 2-AG might function via a CB2-dependent pathway. The present findings show that the HEp-2 LSCC cell line releases the major endocannabinoids AEA and 2-AG, and that their supplementation inhibits tumor cell proliferation in vitro. Thus, cannabinoid ligands might represent novel drug candidates for laryngeal cancers, although functional in vivo studies are required in order to validate their potency. 

Retracted: No

PubPeer comments:
 The cell line used in this paper is HEp-2. The authors use it as a model of laryngeal squamous cell carcinoma (LSCC). They indicate:  The human HEp-2 LSCC cell line was purchased from the ATCC Company (HEp-2, CCl23, USA) . HEp-2 is known since 1968!!!! to be a contaminated HeLa cell line. Furthermore if the the authors did purchase it from ATCC, they can not ignore this as it is clearly stated on the ATCC site on the CCL-23 product page  Tissue: HeLa contaminant  . It is also stated in many other places in the description of this cell line at ATCC. See: https: www.atcc.org Products All CCL-23.aspx Therefore all the conclusions described in this paper are irrelevant to LSCC. It is a shame that Springer can publish such type of studies without checking the origin of this cell line 53 years after it has been shown to be contaminated. See the relevant [Cellosaurus entry] ,