PMID- 30621560 OWN - NLM STAT- MEDLINE DCOM- 20191003 LR - 20191007 IS - 1875-5666 (Electronic) IS - 1566-5240 (Linking) VI - 18 IP - 9 DP - 2018 TI - Determination of Expression Patterns of Seven De-sumoylation Enzymes in Major Ocular Cell Lines. PG - 584-593 LID - 10.2174/1566524019666190107153440 [doi] AB - PURPOSE: Accumulated evidence have well established that protein sumoylation plays multiple roles in various cellular processes. In the vertebrate eye, we and others have demonstrated that sumoylation displays indispensable roles in regulating eye development. Various ocular cell lines including human embryonic cell line (FHL124), the SV40-large T-transformed human lens epithelial cell line (HLE), the SV40-large T-transformed mouse lens epithelial cell line (alphaTN4-1), the rabbit lens epithelial cell line (N/N1003A) and the human retina pigment epithelial cell line (ARPE-19) have been extensively used for studying various cellular functions and disease processes including sumoylation functions, and mechanisms for cataract and age-related macular degeneration (AMD). However, the sumoylation enzyme systems have not been well established. METHODS: FHL124, HLE, alphaTN4-1, N/N1003A and ARPE-19 were cultured in Dulbecco's modified eagle medium (DMEM) containing 10% FBS and 1% penicillin & streptomycin. The expression levels of seven SENP mRNAs were analyzed with qRT-PCR, and the expression levels of seven SENP proteins were detected with Western blot analysis. RESULTS: Using both qRT-PCR and Western blot analysis, we have obtained the followings: 1). The 3 human ocular cell lines, FHL124, HLE and ARPE-19 express all types of SENP mRNA and proteins. 2). In mouse lens epithelial cell line alphaTN4-1, and rabbit lens epithelial cells line N/N1003A, however, only the mRNAs for SENP1, 2, 3, 6 and 7 are expressed. At the protein level, SENP8 was absent in both alphaTN4-1 and N/N1003A cells; 3). Each cell line has different dominant SENP enzymes. For FHL124, SENP3, 5, 7 and 8 proteins are relatively dominant. SENP3, 5 and 6 are the major de-sumoylation enzymes in HLE cells. Different from human lens epithelial cells, FHL124 and HLE, human retina pigment epithelial cells (ARPE-19) have SENP3, 7, and 8 as the dominant forms of de-sumoylation enzymes. For mouse lens epithelial cells, SENP1, 3 and 7 are the major de-sumoylation enzymes. On the other hand, the rabbit lens epithelial cells have SENP1, 2 and 7 as the major isoforms. CONCLUSION: Our results for the first time defined the differential expression patterns of the seven types of de-sumoylation enzymes (SENPs) in 5 major ocular cell lines. These results help to establish the basis for the future study of sumoylation functions and the related mechanisms in vertebrate eye. CI - Copyright(c) Bentham Science Publishers; For any queries, please email at epub@benthamscience.net. FAU - Liu, Yunfei AU - Liu Y AD - The State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, #7 Jinsui Road, Guangzhou, Guangdong 510230, China. FAU - Zhang, Lan AU - Zhang L AD - The State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, #7 Jinsui Road, Guangzhou, Guangdong 510230, China. FAU - Tang, Xiangcheng AU - Tang X AD - The State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, #7 Jinsui Road, Guangzhou, Guangdong 510230, China. FAU - Liu, Fangyuan AU - Liu F AD - The State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, #7 Jinsui Road, Guangzhou, Guangdong 510230, China. FAU - Fu, Jia-Ling AU - Fu JL AD - The State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, #7 Jinsui Road, Guangzhou, Guangdong 510230, China. FAU - Gong, Xiao-Dong AU - Gong XD AD - The State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, #7 Jinsui Road, Guangzhou, Guangdong 510230, China. FAU - Wang, Ling AU - Wang L AD - The State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, #7 Jinsui Road, Guangzhou, Guangdong 510230, China. FAU - Nie, Qian AU - Nie Q AD - The State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, #7 Jinsui Road, Guangzhou, Guangdong 510230, China. FAU - Xiang, Jia-Wen AU - Xiang JW AD - The State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, #7 Jinsui Road, Guangzhou, Guangdong 510230, China. FAU - Xiao, Yuan AU - Xiao Y AD - The State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, #7 Jinsui Road, Guangzhou, Guangdong 510230, China. FAU - Liu, Yizhi AU - Liu Y AD - The State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, #7 Jinsui Road, Guangzhou, Guangdong 510230, China. FAU - Li, David Wan-Cheng AU - Li DW AD - The State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, #7 Jinsui Road, Guangzhou, Guangdong 510230, China. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't PL - Netherlands TA - Curr Mol Med JT - Current molecular medicine JID - 101093076 RN - 0 (Eye Proteins) RN - EC 3.4.22.- (Cysteine Endopeptidases) SB - IM MH - Animals MH - Cell Line MH - Cysteine Endopeptidases/*biosynthesis MH - Eye Proteins/*biosynthesis MH - *Gene Expression Regulation, Enzymologic MH - Humans MH - Lens, Crystalline/*enzymology MH - Mice MH - Rabbits OTO - NOTNLM OT - ARPE-19 OT - De-sumoylation OT - FHL124 OT - HLE OT - N/N1003A OT - SENP. OT - alphaTN4-1 EDAT- 2019/01/10 06:00 MHDA- 2019/10/08 06:00 CRDT- 2019/01/10 06:00 PHST- 2018/09/11 00:00 [received] PHST- 2018/11/21 00:00 [revised] PHST- 2018/12/11 00:00 [accepted] PHST- 2019/01/10 06:00 [pubmed] PHST- 2019/10/08 06:00 [medline] PHST- 2019/01/10 06:00 [entrez] AID - CMM-EPUB-95620 [pii] AID - 10.2174/1566524019666190107153440 [doi] PST - ppublish SO - Curr Mol Med. 2018;18(9):584-593. doi: 10.2174/1566524019666190107153440.