miR-499 promotes immature porcine Sertoli cell growth through the PI3K/AKT pathway by targeting the PTEN gene

in Reproduction
Authors:
Hu Gao College of Animal Science and Technology, Hunan Provincial Key Laboratory for Genetic Improvement of Domestic Animal, Hunan Agricultural University, Hunan, Changsha, China

Search for other papers by Hu Gao in
Current site
Google Scholar
PubMed
Close
,
Bin Chen College of Animal Science and Technology, Hunan Provincial Key Laboratory for Genetic Improvement of Domestic Animal, Hunan Agricultural University, Hunan, Changsha, China

Search for other papers by Bin Chen in
Current site
Google Scholar
PubMed
Close
,
Hui Luo College of Animal Science and Technology, Hunan Provincial Key Laboratory for Genetic Improvement of Domestic Animal, Hunan Agricultural University, Hunan, Changsha, China

Search for other papers by Hui Luo in
Current site
Google Scholar
PubMed
Close
,
Bo Weng College of Animal Science and Technology, Hunan Provincial Key Laboratory for Genetic Improvement of Domestic Animal, Hunan Agricultural University, Hunan, Changsha, China

Search for other papers by Bo Weng in
Current site
Google Scholar
PubMed
Close
,
Xiangwei Tang College of Animal Science and Technology, Hunan Provincial Key Laboratory for Genetic Improvement of Domestic Animal, Hunan Agricultural University, Hunan, Changsha, China

Search for other papers by Xiangwei Tang in
Current site
Google Scholar
PubMed
Close
,
Yao Chen College of Animal Science and Technology, Hunan Provincial Key Laboratory for Genetic Improvement of Domestic Animal, Hunan Agricultural University, Hunan, Changsha, China

Search for other papers by Yao Chen in
Current site
Google Scholar
PubMed
Close
,
Anqi Yang College of Animal Science and Technology, Hunan Provincial Key Laboratory for Genetic Improvement of Domestic Animal, Hunan Agricultural University, Hunan, Changsha, China

Search for other papers by Anqi Yang in
Current site
Google Scholar
PubMed
Close
, and
Maoliang Ran College of Animal Science and Technology, Hunan Provincial Key Laboratory for Genetic Improvement of Domestic Animal, Hunan Agricultural University, Hunan, Changsha, China

Search for other papers by Maoliang Ran in
Current site
Google Scholar
PubMed
Close

Correspondence should be addressed to M Ran or B Chen; Email: ranmaoliang0903@126.com or chenbin7586@126.com
Restricted access
Rent on DeepDyve

Sign up for journal news

Sertoli cells are indispensable for normal spermatogenesis, and increasing evidence has shown that miRNAs participate in the regulation of Sertoli cell growth. However, the functions and regulatory mechanisms of miRNAs in Sertoli cells of domestic animals have not been fully investigated. In the present study, we mainly investigated the regulatory roles of miR-499 in immature porcine Sertoli cells. The results showed that miR-499 was mainly located in the basement section of seminiferous tubules of prepubertal porcine testicular tissue. Overexpression of miR-499 promoted cell proliferation and inhibited apoptosis, whereas miR-499 inhibition resulted in the opposite effect. The PTEN gene was directly targeted by miR-499, and the expression of mRNA and protein was also negatively regulated by miR-499 in immature porcine Sertoli cells. siRNA-induced PTEN knockdown resulted in a similar effect as an overexpression of miR-499 and abolished the effects of miR-499 inhibition on immature porcine Sertoli cells. Moreover, both miR-499 overexpression and the PTEN knockdown activated the PI3K/AKT signaling pathway, whereas inhibition of the PI3K/AKT signaling pathway caused immature porcine Sertoli cell apoptosis and inhibited cell proliferation. Overall, miR-499 promotes proliferation and inhibits apoptosis in immature porcine Sertoli cells through the PI3K/AKT pathway by targeting the PTEN gene. This study provides novel insights into the effects of miR-499 in spermatogenesis through the regulation of immature Sertoli cell proliferation and apoptosis.

Supplementary Materials

    • Figure S1 The transfection efficiency of miR-499 and PTEN. (A) The transfection efficiency of immature porcine Sertoli cells was measured using a riboTRACERTM Fluorescent Oligo (green). The representative images are presented. Scale bar = 50 mm. (B) Immature porcine Sertoli cells were transfected with mimic NC, miR-499 mimic, inhibitor NC, or miR-499 inhibitor. The miR-499 expression was measured using the qPCR assay (n = 3). U6 was used as the internal control. (C) Cells were transfected with siRNA NC or PTEN siRNA. PTEN expression was measured using qPCR and western blot assays (n = 3). pig-TBP and β-actin were used as internal controls, respectively. Data are presented as the mean ± S.D. *P<0.05 and **P<0.01.
    • Table S1 Table S1 Synthetic oligo sequences
    • Table S2 The sequences of the primers used in this study.

 

  • Collapse
  • Expand