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PU.1 antisense lnc RNA against its m RNA translation promotes adipogenesis in porcine preadipocytes
Author(s) -
Wei N.,
Wang Y.,
Xu R.X.,
Wang G.Q.,
Xiong Y.,
Yu T.Y.,
Yang G.S.,
Pang W.J.
Publication year - 2015
Publication title -
animal genetics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.756
H-Index - 81
eISSN - 1365-2052
pISSN - 0268-9146
DOI - 10.1111/age.12275
Subject(s) - adipogenesis , gene knockdown , adipose tissue , biology , adipose triglyceride lipase , peroxisome proliferator activated receptor gamma , messenger rna , fatty acid synthase , endocrinology , medicine , peroxisome proliferator activated receptor , receptor , gene , biochemistry , lipid metabolism , lipolysis
Summary Antisense long non‐coding RNA s ( AS lnc RNA s) play important roles in refined regulation of animal gene expression. However, their functions and molecular mechanisms for domestic animal adipogenesis are largely unknown. Here, we found a novel AS lnc RNA transcribed from the porcine PU.1 gene (also known as SPI 1 ) by strand‐specific RT ‐ PCR . Results showed that PU.1 AS lnc RNA was expressed and generally lower than the level of PU.1 m RNA in porcine subcutaneous adipose, heart, liver, spleen, lympha, skeletal muscle and kidney tissues. We further found that the levels of PU.1 m RNA and PU.1 protein were significantly lower in subcutaneous and intermuscular adipose than in mesenteric and greater omentum adipose, whereas the levels of PU.1 AS lnc RNA showed no difference in porcine adipose tissues from four different parts of the body. During porcine adipogenesis, levels of PU.1 m RNA increased at day 2 and then gradually decreased. Meanwhile, PU.1 AS lnc RNA exhibited an expression trend similar to PU.1 m RNA but sharply decreased after day 2. Interestingly, PU.1 protein level rose during differentiation. In addition, at day 6 after differentiation, knockdown of endogenous PU.1 promoted adipogenesis, whereas knockdown of endogenous PU.1 AS lnc RNA had the opposite effect. Moreover, peroxisome proliferator‐activated receptor gamma ( PPARG ) and fatty acid synthase ( FASN ) were significantly upregulated in the PU.1 sh RNA treatment group ( P < 0.05), whereas they were downregulated in the PU.1 AS sh RNA treatment group ( P < 0.05). Adipose triglyceride lipase [ ATGL ; also known as patatin‐like phospholipase domain containing 2 ( PNPLA 2 )] and hormone‐sensitive lipase [ HSL; also known as lipase, hormone‐sensitive ( LIPE )] contrasted with PPARG and FASN . Finally, the PU.1 m RNA / PU.1 AS lnc RNA duplex was detected by an endogenous ribonuclease protection assay combined with RT ‐ PCR . Based on the above results, we suggest that PU.1 AS lnc RNA (vs. its m RNA translation) promotes adipogenesis through the formation of a sense–antisense RNA duplex with PU.1 m RNA .