HOXA9 gene inhibits proliferation and differentiation and promotes apoptosis of bovine preadipocytes

Abstract Background Hox gene family is an important transcription factor that regulates cell process, and plays a role in the process of adipocytes differentiation and fat deposition. Previous transcriptome sequencing studies have indicated that the Homeobox A9 gene (HOXA9) is a candidate gene for r...

Full description

Bibliographic Details
Main Authors: Lixia He, Xue Feng, Chunli Hu, Shuang Liu, Hui Sheng, Bei Cai, Yun Ma
Format: Article
Language:English
Published: BMC 2024-04-01
Series:BMC Genomics
Subjects:
Online Access:https://doi.org/10.1186/s12864-024-10231-3
_version_ 1827284465237360640
author Lixia He
Xue Feng
Chunli Hu
Shuang Liu
Hui Sheng
Bei Cai
Yun Ma
author_facet Lixia He
Xue Feng
Chunli Hu
Shuang Liu
Hui Sheng
Bei Cai
Yun Ma
author_sort Lixia He
collection DOAJ
description Abstract Background Hox gene family is an important transcription factor that regulates cell process, and plays a role in the process of adipocytes differentiation and fat deposition. Previous transcriptome sequencing studies have indicated that the Homeobox A9 gene (HOXA9) is a candidate gene for regulating the process of bovine lipid metabolism, but the function and specific mechanism of action remain unclear. Therefore, this study aims to explore the role of HOXA9 in the proliferation, differentiation and apoptosis of bovine preadipocytes through gain-of-function and lose-of-function. Result It found HOXA9 highly expressed in bovine adipose tissue, and its expression level changed significantly during adipocytes differentiation process. It gave a hint that HOXA9 may be involved in the process of bovine lipid metabolism. The results of HOXA9 gain-of-function experiments indicated that HOXA9 appeared to act as a negative regulator not only in the differentiation but also in the proliferation of bovine preadipocytes, which is mainly reflected that overexpression of HOXA9 down-regulate the mRNA and protein expression level of PPARγ, CEBPα and FABP4 (P < 0.05). The mRNA expression level of CDK1, CDK2, PCNA, CCNA2, CCNB1, CCND1 and CCNE2, as well as the protein expression of CDK2 also significantly decreased. The decrease of lipid droplets content was the main characteristic of the phenotype (P < 0.01), which further supported the evidence that HOXA9 was a negative regulator of preadipocytes differentiation. The decrease of cell proliferation rate and EdU positive rate, as well as the limitation of transition of preadipocytes from G0/G1 phase to S phase also provided evidence for the inhibition of proliferation. Apart from this above, we noted an interesting phenomenon that overexpression of HOXA9 showed in a significant upregulation of both mRNA and protein level of apoptosis markers, accompanied by a significant increase in cell apoptosis rate. These data led us not to refute the fact that HOXA9 played an active regulatory role in apoptosis. HOXA9 loss-of-function experiments, however, yielded the opposite results. Considering that HOXA9 acts as a transcription factor, we predicted its target genes. Dual luciferase reporter assay system indicated that overexpression of HOXA9 inhibits activity of PCNA promoter. Conclusion Taken together, we demonstrated for the first time that HOXA9 played a role as a negative regulatory factor in the differentiation and proliferation of preadipocytes, but played a positive regulatory role in apoptosis, and it may play a regulatory role by targeting PCNA. This study provides basic data for further exploring the regulatory network of intramuscular fat deposition in bovine.
first_indexed 2024-04-24T09:56:58Z
format Article
id doaj.art-bcf710186ba64a5bb5ff9d6d70983da7
institution Directory Open Access Journal
issn 1471-2164
language English
last_indexed 2024-04-24T09:56:58Z
publishDate 2024-04-01
publisher BMC
record_format Article
series BMC Genomics
spelling doaj.art-bcf710186ba64a5bb5ff9d6d70983da72024-04-14T11:08:44ZengBMCBMC Genomics1471-21642024-04-0125111310.1186/s12864-024-10231-3HOXA9 gene inhibits proliferation and differentiation and promotes apoptosis of bovine preadipocytesLixia He0Xue Feng1Chunli Hu2Shuang Liu3Hui Sheng4Bei Cai5Yun Ma6College of Animal Science and Technology, Key Laboratory of Ruminant Molecular and Cellular Breeding of Ningxia Hui Autonomous Region, Ningxia UniversityCollege of Animal Science and Technology, Key Laboratory of Ruminant Molecular and Cellular Breeding of Ningxia Hui Autonomous Region, Ningxia UniversityCollege of Animal Science and Technology, Key Laboratory of Ruminant Molecular and Cellular Breeding of Ningxia Hui Autonomous Region, Ningxia UniversityCollege of Animal Science and Technology, Key Laboratory of Ruminant Molecular and Cellular Breeding of Ningxia Hui Autonomous Region, Ningxia UniversityCollege of Animal Science and Technology, Key Laboratory of Ruminant Molecular and Cellular Breeding of Ningxia Hui Autonomous Region, Ningxia UniversityCollege of Animal Science and Technology, Key Laboratory of Ruminant Molecular and Cellular Breeding of Ningxia Hui Autonomous Region, Ningxia UniversityCollege of Animal Science and Technology, Key Laboratory of Ruminant Molecular and Cellular Breeding of Ningxia Hui Autonomous Region, Ningxia UniversityAbstract Background Hox gene family is an important transcription factor that regulates cell process, and plays a role in the process of adipocytes differentiation and fat deposition. Previous transcriptome sequencing studies have indicated that the Homeobox A9 gene (HOXA9) is a candidate gene for regulating the process of bovine lipid metabolism, but the function and specific mechanism of action remain unclear. Therefore, this study aims to explore the role of HOXA9 in the proliferation, differentiation and apoptosis of bovine preadipocytes through gain-of-function and lose-of-function. Result It found HOXA9 highly expressed in bovine adipose tissue, and its expression level changed significantly during adipocytes differentiation process. It gave a hint that HOXA9 may be involved in the process of bovine lipid metabolism. The results of HOXA9 gain-of-function experiments indicated that HOXA9 appeared to act as a negative regulator not only in the differentiation but also in the proliferation of bovine preadipocytes, which is mainly reflected that overexpression of HOXA9 down-regulate the mRNA and protein expression level of PPARγ, CEBPα and FABP4 (P < 0.05). The mRNA expression level of CDK1, CDK2, PCNA, CCNA2, CCNB1, CCND1 and CCNE2, as well as the protein expression of CDK2 also significantly decreased. The decrease of lipid droplets content was the main characteristic of the phenotype (P < 0.01), which further supported the evidence that HOXA9 was a negative regulator of preadipocytes differentiation. The decrease of cell proliferation rate and EdU positive rate, as well as the limitation of transition of preadipocytes from G0/G1 phase to S phase also provided evidence for the inhibition of proliferation. Apart from this above, we noted an interesting phenomenon that overexpression of HOXA9 showed in a significant upregulation of both mRNA and protein level of apoptosis markers, accompanied by a significant increase in cell apoptosis rate. These data led us not to refute the fact that HOXA9 played an active regulatory role in apoptosis. HOXA9 loss-of-function experiments, however, yielded the opposite results. Considering that HOXA9 acts as a transcription factor, we predicted its target genes. Dual luciferase reporter assay system indicated that overexpression of HOXA9 inhibits activity of PCNA promoter. Conclusion Taken together, we demonstrated for the first time that HOXA9 played a role as a negative regulatory factor in the differentiation and proliferation of preadipocytes, but played a positive regulatory role in apoptosis, and it may play a regulatory role by targeting PCNA. This study provides basic data for further exploring the regulatory network of intramuscular fat deposition in bovine.https://doi.org/10.1186/s12864-024-10231-3HOXA9BovineFat depositionDifferentiationProliferation
spellingShingle Lixia He
Xue Feng
Chunli Hu
Shuang Liu
Hui Sheng
Bei Cai
Yun Ma
HOXA9 gene inhibits proliferation and differentiation and promotes apoptosis of bovine preadipocytes
BMC Genomics
HOXA9
Bovine
Fat deposition
Differentiation
Proliferation
title HOXA9 gene inhibits proliferation and differentiation and promotes apoptosis of bovine preadipocytes
title_full HOXA9 gene inhibits proliferation and differentiation and promotes apoptosis of bovine preadipocytes
title_fullStr HOXA9 gene inhibits proliferation and differentiation and promotes apoptosis of bovine preadipocytes
title_full_unstemmed HOXA9 gene inhibits proliferation and differentiation and promotes apoptosis of bovine preadipocytes
title_short HOXA9 gene inhibits proliferation and differentiation and promotes apoptosis of bovine preadipocytes
title_sort hoxa9 gene inhibits proliferation and differentiation and promotes apoptosis of bovine preadipocytes
topic HOXA9
Bovine
Fat deposition
Differentiation
Proliferation
url https://doi.org/10.1186/s12864-024-10231-3
work_keys_str_mv AT lixiahe hoxa9geneinhibitsproliferationanddifferentiationandpromotesapoptosisofbovinepreadipocytes
AT xuefeng hoxa9geneinhibitsproliferationanddifferentiationandpromotesapoptosisofbovinepreadipocytes
AT chunlihu hoxa9geneinhibitsproliferationanddifferentiationandpromotesapoptosisofbovinepreadipocytes
AT shuangliu hoxa9geneinhibitsproliferationanddifferentiationandpromotesapoptosisofbovinepreadipocytes
AT huisheng hoxa9geneinhibitsproliferationanddifferentiationandpromotesapoptosisofbovinepreadipocytes
AT beicai hoxa9geneinhibitsproliferationanddifferentiationandpromotesapoptosisofbovinepreadipocytes
AT yunma hoxa9geneinhibitsproliferationanddifferentiationandpromotesapoptosisofbovinepreadipocytes