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Role of miR-511 in the Regulation of OATP1B1 Expression by Free Fatty Acid.

Peng JF, Liu L, Guo CX, Liu SK, Chen XP, Huang LH, Xiang H, Huang ZJ, Yuan H, Yang GP - Biomol Ther (Seoul) (2015)

Bottom Line: MicroRNAs (miRNAs) are a family of non-coding RNA that are able to adjust the expression of many proteins, including ATP-binding cassette transporter and organic cation transporter.We also proved that SLCO1B1 is the target gene of miR-511 with a bioinformatics analysis and using the dual luciferase reporter assay.Furthermore, the expressions of SLCO1B1 and OATP1B1 decreased if transfecting Chang liver cells with miR-511, but did not increase when transfecting the inhibitors of miR-511 into steatosis cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacy, The Third Xiangya Hospital, Central South University, Changsha, Hunan 410000 ; Center of Clinical Pharmacology, The Third Xiangya Hospital, Central South University, Changsha, Hunan 410013.

ABSTRACT
MicroRNAs (miRNAs) are a family of non-coding RNA that are able to adjust the expression of many proteins, including ATP-binding cassette transporter and organic cation transporter. We sought to evaluate the effect of miR-511 on the regulation of OATP1B1 expression by free fatty acids. When using free fatty acids to stimulate Chang liver cells, we found that the expression of miR-511 increased significantly while the expression of OATP1B1 decreased. We also proved that SLCO1B1 is the target gene of miR-511 with a bioinformatics analysis and using the dual luciferase reporter assay. Furthermore, the expressions of SLCO1B1 and OATP1B1 decreased if transfecting Chang liver cells with miR-511, but did not increase when transfecting the inhibitors of miR-511 into steatosis cells. Our study indicates that miR-511 may play an important role in the regulation of OATP1B1 expression by free fatty acids.

No MeSH data available.


Related in: MedlinePlus

SLCO1B1 mRNA and protein expression in the steatosis cells. (A) The SLCO1B1 mRNA level was determined by real time Q-PCR and normalized with the GAPDH mRNA level. (B) The OATP1B1 protein level was determined using Western blot analysis and normalized with the GAPDH protein level. (C) The gray values of OATP1B1 were measured using Image J software and normalized with GAPDH. The control was treated only with 1% BSA, while steatosis cells were exposed to 1mM FFA for 24 h. Each column represented the mean ± SD of three independent experiments. *p<0.05 versus control.
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f3-bt-23-400: SLCO1B1 mRNA and protein expression in the steatosis cells. (A) The SLCO1B1 mRNA level was determined by real time Q-PCR and normalized with the GAPDH mRNA level. (B) The OATP1B1 protein level was determined using Western blot analysis and normalized with the GAPDH protein level. (C) The gray values of OATP1B1 were measured using Image J software and normalized with GAPDH. The control was treated only with 1% BSA, while steatosis cells were exposed to 1mM FFA for 24 h. Each column represented the mean ± SD of three independent experiments. *p<0.05 versus control.

Mentions: The SLCO1B1 mRNA level was first examined using real time RT-PCR and then OATP1B1 protein expression was detected in the steatosis cells. As shown in Fig. 3A, a statistically significant decrease was observed in the steatosis cells versus the Chang liver cells (p=0.004). The gray values were measured using Image J software and OATP1B1 was normalized by GAPDH. Likewise, the OATP1B1 protein level decreased (Fig. 3B, 3C). Furthermore, the result of immunocytochemistry also verified that FFA decreased the expression of OATP1B1 (data not shown).


Role of miR-511 in the Regulation of OATP1B1 Expression by Free Fatty Acid.

Peng JF, Liu L, Guo CX, Liu SK, Chen XP, Huang LH, Xiang H, Huang ZJ, Yuan H, Yang GP - Biomol Ther (Seoul) (2015)

SLCO1B1 mRNA and protein expression in the steatosis cells. (A) The SLCO1B1 mRNA level was determined by real time Q-PCR and normalized with the GAPDH mRNA level. (B) The OATP1B1 protein level was determined using Western blot analysis and normalized with the GAPDH protein level. (C) The gray values of OATP1B1 were measured using Image J software and normalized with GAPDH. The control was treated only with 1% BSA, while steatosis cells were exposed to 1mM FFA for 24 h. Each column represented the mean ± SD of three independent experiments. *p<0.05 versus control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4556198&req=5

f3-bt-23-400: SLCO1B1 mRNA and protein expression in the steatosis cells. (A) The SLCO1B1 mRNA level was determined by real time Q-PCR and normalized with the GAPDH mRNA level. (B) The OATP1B1 protein level was determined using Western blot analysis and normalized with the GAPDH protein level. (C) The gray values of OATP1B1 were measured using Image J software and normalized with GAPDH. The control was treated only with 1% BSA, while steatosis cells were exposed to 1mM FFA for 24 h. Each column represented the mean ± SD of three independent experiments. *p<0.05 versus control.
Mentions: The SLCO1B1 mRNA level was first examined using real time RT-PCR and then OATP1B1 protein expression was detected in the steatosis cells. As shown in Fig. 3A, a statistically significant decrease was observed in the steatosis cells versus the Chang liver cells (p=0.004). The gray values were measured using Image J software and OATP1B1 was normalized by GAPDH. Likewise, the OATP1B1 protein level decreased (Fig. 3B, 3C). Furthermore, the result of immunocytochemistry also verified that FFA decreased the expression of OATP1B1 (data not shown).

Bottom Line: MicroRNAs (miRNAs) are a family of non-coding RNA that are able to adjust the expression of many proteins, including ATP-binding cassette transporter and organic cation transporter.We also proved that SLCO1B1 is the target gene of miR-511 with a bioinformatics analysis and using the dual luciferase reporter assay.Furthermore, the expressions of SLCO1B1 and OATP1B1 decreased if transfecting Chang liver cells with miR-511, but did not increase when transfecting the inhibitors of miR-511 into steatosis cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacy, The Third Xiangya Hospital, Central South University, Changsha, Hunan 410000 ; Center of Clinical Pharmacology, The Third Xiangya Hospital, Central South University, Changsha, Hunan 410013.

ABSTRACT
MicroRNAs (miRNAs) are a family of non-coding RNA that are able to adjust the expression of many proteins, including ATP-binding cassette transporter and organic cation transporter. We sought to evaluate the effect of miR-511 on the regulation of OATP1B1 expression by free fatty acids. When using free fatty acids to stimulate Chang liver cells, we found that the expression of miR-511 increased significantly while the expression of OATP1B1 decreased. We also proved that SLCO1B1 is the target gene of miR-511 with a bioinformatics analysis and using the dual luciferase reporter assay. Furthermore, the expressions of SLCO1B1 and OATP1B1 decreased if transfecting Chang liver cells with miR-511, but did not increase when transfecting the inhibitors of miR-511 into steatosis cells. Our study indicates that miR-511 may play an important role in the regulation of OATP1B1 expression by free fatty acids.

No MeSH data available.


Related in: MedlinePlus