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miR-541 Contributes to Microcystin-LR-Induced Reproductive Toxicity through Regulating the Expression of p15 in Mice

View Article: PubMed Central - PubMed

ABSTRACT

Microcystin-leucine arginine (MC-LR) is a harmful cyanotoxin produced by cyanobacteria. MC-LR can exert endocrine-disrupting activities in many organisms. We have previously demonstrated that MC-LR exerts both acute and chronic reproductive toxicity in male mice, resulting in a decline in sperm quality and damage to testicular structure. Moreover, we also observed extensive alterations in a panel of microRNAs in spermatogonial cells after exposure to MC-LR. In this study, we have confirmed that miR-541 was significantly increased both in GC-1 cells (in vitro) and in mouse testes (in vivo) after exposure to MC-LR. Our data support that p15 was the target gene of miR-541. Increase in miR-541 led to a reduction of p15 and murine double minute2 (MDM2), promoting the activation of p53 signaling and MC-LR-mediated cell apoptosis. Moreover, cells responded to MC-LR with reduced viability and increased apoptosis. Consistently, inhibiting miR-541 could upregulate the expression of p15 and MDM2, resulting in the downregulation of phospho-p53. Downregulation of miR-541 promoted cell viability by reducing MC-LR-induced cell apoptosis. In conclusion, we demonstrate here a crucial role for miR-541 in MC-LR-induced toxic effects on the reproductive system, in an attempt to provide a rational strategy for the diagnosis and treatment of MC-LR-induced impairment in the reproductive system.

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Related in: MedlinePlus

The effects of MC-LR on the expression of miR-541 and p15 in mouse testes. (A,B) Mice were intraperitoneally injected with various concentrations of MC-LR. The mRNA levels of miR-541 (A) and p15 (B) in testes were measured with qRT-PCR; (C) The protein levels of p15 in testicular tissues were measured by Western blot. The expression of p15 was quantified with ImageJ (right panels; n = 3). β-actin was used as a loading control. Results are expressed as means ± S.D. * p < 0.05.
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toxins-08-00260-f004: The effects of MC-LR on the expression of miR-541 and p15 in mouse testes. (A,B) Mice were intraperitoneally injected with various concentrations of MC-LR. The mRNA levels of miR-541 (A) and p15 (B) in testes were measured with qRT-PCR; (C) The protein levels of p15 in testicular tissues were measured by Western blot. The expression of p15 was quantified with ImageJ (right panels; n = 3). β-actin was used as a loading control. Results are expressed as means ± S.D. * p < 0.05.

Mentions: The expression of miR-541 and p15 was determined in testicular tissue from either control mice or mice treated with MC-LR. miR-541 was significantly upregulated when higher concentrations of MC-LR (15 μg/kg and 30 μg/kg) were delivered (Figure 4A). The mRNA levels of p15 remained unchanged (Figure 4B) and the protein levels of p15 were significantly downregulated (Figure 4C). These results were consistent with our in vitro findings.


miR-541 Contributes to Microcystin-LR-Induced Reproductive Toxicity through Regulating the Expression of p15 in Mice
The effects of MC-LR on the expression of miR-541 and p15 in mouse testes. (A,B) Mice were intraperitoneally injected with various concentrations of MC-LR. The mRNA levels of miR-541 (A) and p15 (B) in testes were measured with qRT-PCR; (C) The protein levels of p15 in testicular tissues were measured by Western blot. The expression of p15 was quantified with ImageJ (right panels; n = 3). β-actin was used as a loading control. Results are expressed as means ± S.D. * p < 0.05.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5037486&req=5

toxins-08-00260-f004: The effects of MC-LR on the expression of miR-541 and p15 in mouse testes. (A,B) Mice were intraperitoneally injected with various concentrations of MC-LR. The mRNA levels of miR-541 (A) and p15 (B) in testes were measured with qRT-PCR; (C) The protein levels of p15 in testicular tissues were measured by Western blot. The expression of p15 was quantified with ImageJ (right panels; n = 3). β-actin was used as a loading control. Results are expressed as means ± S.D. * p < 0.05.
Mentions: The expression of miR-541 and p15 was determined in testicular tissue from either control mice or mice treated with MC-LR. miR-541 was significantly upregulated when higher concentrations of MC-LR (15 μg/kg and 30 μg/kg) were delivered (Figure 4A). The mRNA levels of p15 remained unchanged (Figure 4B) and the protein levels of p15 were significantly downregulated (Figure 4C). These results were consistent with our in vitro findings.

View Article: PubMed Central - PubMed

ABSTRACT

Microcystin-leucine arginine (MC-LR) is a harmful cyanotoxin produced by cyanobacteria. MC-LR can exert endocrine-disrupting activities in many organisms. We have previously demonstrated that MC-LR exerts both acute and chronic reproductive toxicity in male mice, resulting in a decline in sperm quality and damage to testicular structure. Moreover, we also observed extensive alterations in a panel of microRNAs in spermatogonial cells after exposure to MC-LR. In this study, we have confirmed that miR-541 was significantly increased both in GC-1 cells (in vitro) and in mouse testes (in vivo) after exposure to MC-LR. Our data support that p15 was the target gene of miR-541. Increase in miR-541 led to a reduction of p15 and murine double minute2 (MDM2), promoting the activation of p53 signaling and MC-LR-mediated cell apoptosis. Moreover, cells responded to MC-LR with reduced viability and increased apoptosis. Consistently, inhibiting miR-541 could upregulate the expression of p15 and MDM2, resulting in the downregulation of phospho-p53. Downregulation of miR-541 promoted cell viability by reducing MC-LR-induced cell apoptosis. In conclusion, we demonstrate here a crucial role for miR-541 in MC-LR-induced toxic effects on the reproductive system, in an attempt to provide a rational strategy for the diagnosis and treatment of MC-LR-induced impairment in the reproductive system.

No MeSH data available.


Related in: MedlinePlus