Effect of lncRNA2099 on Proliferation and Differentiation of Porcine Preadipocytes

doi:10.7668/hbnxb.20194328

Abstract

Abstract:

To investigate the effect of lncRNA2099 on the proliferation and differentiation of porcine preadipocytes,the qRT-PCR was used to detect the expression level of lncRNA2099 in heart,liver,spleen,lung,kidney,back muscle,back fat and proliferation and differentiation stages of porcine preadipocyte.The nucleoplasmic separation technique was used to determine the expression location of lncRNA2099 in adipocyte.The eukaryotic expression vector pcDNA3.1-lncRNA2099 was constructed and qRT-PCR was used to detect its impact on the expression of marker genes related to adipocyte proliferation and differentiation after over expression in adipocyte.The results showed that lncRNA2099 was highly expressed in kidney and back fat,but not in back muscle.The nucleoplasmic localization results showed that lncRNA2099 was expressed in both nucleus and cytoplasm;the expression levels of proliferation marker gene P21 and lipogenic differentiation marker gene LPL significantly increased after overexpression of lncRNA2099 in adipocytes.lncRNA2099 may act as a transcriptional regulator to inhibit proliferation and promote lipogenic differentiation in porcine preadipocytes.

Key words: Pig, lncRNA2099, Preadipocytes, Proliferation, Lipogenic differentiation

ZHANG Dongjie, MA Shouzheng, WANG Liang, LIU Di. Effect of lncRNA2099 on Proliferation and Differentiation of Porcine Preadipocytes[J]. Acta Agriculturae Boreali-Sinica, 2024, 39(1): 184-190. doi: 10.7668/hbnxb.20194328.

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http://www.hbnxb.net/EN/Y2024/V39/I1/184

Figures/Tables 7

Tab.1 Primer sequences for qRT-PCR assays

基因 Genes	引物序列(5'—3') Primer sequence(5'—3')		GenBank登录号 GenBank accession number
GAPDH	F:GGTGAAGGTCGGAGTGAACG	R:CTCGCTCCTGGAAGATGGTG	NC_010447.5
lncRNA2099	F:ATGGATGTGAGTCAGATTCGTT	R:AAAAGATGTCTGCATTGTAATC	XR_002342099
Neat1	F:AGAGTGATGCCTGGTAGATGG	R:GTGGCTTCCTGTGAGTATTGG	NC_000011.10
CDK4	F:GCCCTCAAGAGCGTAAGA	R:GCAGTGGCACAGACATCC	NC_010447.5
CyclinB	F:AATCCCTTCTTGTGGTTA	R:CTTAGATGTGGCATACTTG	NC_010458.4
CyclinD	F:TACACCGACAACTCCATCCG	R:GAGGGCGGGTTGGAAATGAA	NC_010447.5
CyclinE	F:CAGAGCAGCGAGCAGGAGC	R:GCAAGCTGCTTCCACACCACAT	NC_010446.5
P21	F:ACGTCTCAGGAGGACCATGT	R:AGAAGATCAGCCGGCGTTTG	NC_010461.5
LPL	F:AACTTGTGGCTGCCCTATAC	R:GTGACCCTCTGGTGAATGTG	NC_010456.5
C/EBPα	F:TAGACAAGAACAGCAACGAG	R:ACCTTCTGTTGAGTCTCCACG	NC_010448.4
PPARγ	F:CATTCGCATCTTTCAGGG	R:GGACGCCATACTTTAGG	NC_010455.5
FABP4	F:GAAAGTCAAGAGCACCAT	R:ATACATTCCACCACCAAC	NM_001002817

Fig.1 PCR amplification results of pig lncRNA2099 M.2 000 bp Marker; 1.lncRNA2099 fragment.

Fig.2 Tissue expression profile of lncRNA2099 Different lowercase letters indicate significant differences in different tissue (P<0.05).The same as Fig.6.

Fig.3 Results of nucleoplasmic RNA isolation and detection of lncRNA2099

Fig.4 Temporal expression of lncRNA2099 in proliferating and differentiating preadipocytes of pigs

Fig.5 Proliferation and differentiation characteristics of porcine preadipocytes

Fig.6 Detection of the effect of overexpression of lncRNA2099 A.lncRNA2099 overexpression plasmid double digestion:M1.10 000 bp Marker,P1.Control plasmid,P2.Double digestion 2099 overexpression vector,M2.2000 bp Marker;B.lncRNA2099 overexpression efficiency;C.Changes in cell proliferation and differentiation marker gene expression after overexpression of lncRNA2099.

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