Fan Lu1, Dongsheng Yan, Xiangtian Zhou, Dan-Ning Hu, Jia Qu. 1. Eye Hospital, School of Ophthalmology and Optometry, Wenzhou Medical College and Key Laboratory of Vision Science, Ministry of Health PR China, Wenzhou China.
Abstract
PURPOSE: Controversy exists over melanogenesis of adult human RPE cells in vitro. This study investigated melanin content and production and expression of tyrosinase (TYR), tyrosinase-related-protein-1 (TRP1), tyrosinase-related-protein-2 (TRP2), and P gene in cultured human RPE cells and uveal melanoma cells. METHODS: RPE cells were isolated and cultured from three adult donor eyes. A continuous human uveal melanoma cell line was established from primary choroidal melanoma. Melanin content and production were measured, and the expression of TYR, TRP1, TRP2, and P gene at the mRNA and protein levels were detected by RT-PCR and western blot, respectively. RESULTS: Melanin content per cell of cultured RPE decreased rapidly and in proportion to cell division. No melanin production could be demonstrated in any passages. In cultured RPE cells, mRNA expression of TYR, TRP1, TRP2, and P-gene and protein expression of TYR, TRP1, and TRP2 could not be detected. In uveal melanoma cells, melanin content per cell remained stable, and melanin production could be detected in each passage. Expression of mRNA of TYR, TRP1, TRP2, and P-gene and protein of TYR, TRP1, and TRP2 could be detected in melanoma cells. CONCLUSIONS: Human RPE cells under standard culture circumstances do not produce melanin and do not express the four key genes required in melanin biosynthesis pathway. In contrast, human uveal melanoma cells produce melanin and express all of these melanogenic genes in vitro.
PURPOSE: Controversy exists over melanogenesis of adult human RPE cells in vitro. This study investigated melanin content and production and expression of tyrosinase (TYR), tyrosinase-related-protein-1 (TRP1), tyrosinase-related-protein-2 (TRP2), and P gene in cultured human RPE cells and uveal melanoma cells. METHODS: RPE cells were isolated and cultured from three adult donor eyes. A continuous humanuveal melanoma cell line was established from primary choroidal melanoma. Melanin content and production were measured, and the expression of TYR, TRP1, TRP2, and P gene at the mRNA and protein levels were detected by RT-PCR and western blot, respectively. RESULTS:Melanin content per cell of cultured RPE decreased rapidly and in proportion to cell division. No melanin production could be demonstrated in any passages. In cultured RPE cells, mRNA expression of TYR, TRP1, TRP2, and P-gene and protein expression of TYR, TRP1, and TRP2 could not be detected. In uveal melanoma cells, melanin content per cell remained stable, and melanin production could be detected in each passage. Expression of mRNA of TYR, TRP1, TRP2, and P-gene and protein of TYR, TRP1, and TRP2 could be detected in melanoma cells. CONCLUSIONS:Human RPE cells under standard culture circumstances do not produce melanin and do not express the four key genes required in melanin biosynthesis pathway. In contrast, humanuveal melanoma cells produce melanin and express all of these melanogenic genes in vitro.
Authors: J Jha; M K Singh; L Singh; N Pushker; N Lomi; R Meel; K Chosdol; S Sen; S Bakhshi; S Kashyap Journal: Clin Transl Oncol Date: 2021-04-03 Impact factor: 3.405
Authors: L Pasovic; T P Utheim; S Reppe; A Z Khan; C J Jackson; B Thiede; J P Berg; E B Messelt; J R Eidet Journal: Sci Rep Date: 2018-04-09 Impact factor: 4.379