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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

The lipid accumulation. (A, B, C, D) Representative microscopy photograph. Chang liver cells were exposed to 2 mM, 1 mM, 0.5 mM FFA and 1% BSA for 24 h. The control cells were treated only with 1% BSA. (E) Cellular viability after incubating with different FFAs for various durations. Chang liver cells were incubated with 2 mM, 1 mM, and 0.5 mM FFA for 12 h, 24 h and 48 h respectively. The control cells were only treated with 1% BSA. Single points included the means of at least three independent experiments. (F) Effects of different fatty acids on triglyceride accumulation. The ratio of triglyceride accumulation to the total protein amount (TG/protein, mg/g) was evaluated as the concentration of TG in cell lysate after radio immunoprecipitation. The column represented the mean ± SD of three independent experiments. *p<0.05 versus control.
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f1-bt-23-400: The lipid accumulation. (A, B, C, D) Representative microscopy photograph. Chang liver cells were exposed to 2 mM, 1 mM, 0.5 mM FFA and 1% BSA for 24 h. The control cells were treated only with 1% BSA. (E) Cellular viability after incubating with different FFAs for various durations. Chang liver cells were incubated with 2 mM, 1 mM, and 0.5 mM FFA for 12 h, 24 h and 48 h respectively. The control cells were only treated with 1% BSA. Single points included the means of at least three independent experiments. (F) Effects of different fatty acids on triglyceride accumulation. The ratio of triglyceride accumulation to the total protein amount (TG/protein, mg/g) was evaluated as the concentration of TG in cell lysate after radio immunoprecipitation. The column represented the mean ± SD of three independent experiments. *p<0.05 versus control.

Mentions: After the Chang liver cells were exposed to successively graded concentrations of FFA (2 mM, 1 mM, 0.5 mM) for 24 h, oil red O staining was performed and the results shown in Fig. 1 A∼D indicating the lipid accumulation was FFA concentration-dependent. The cellular viability was tested using the methyl thiazolyl tetrazolium (MTT) method and the ratio of triglyceride (TG) accumulation to the total protein amount (TG/protein, mg/g) was assessed to confirm lipid accumulation. As shown in Fig. 1E, 1F, the cellular viability and TG/protein ratio exhibited a time-dependent and FFA concentration-dependent phenomenon. For overall considerations of the cellular viability and the lipid accumulation, the optimal incubation time was 24 hours and the optimal incubation concentration of FFA was 1 mM.


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)

The lipid accumulation. (A, B, C, D) Representative microscopy photograph. Chang liver cells were exposed to 2 mM, 1 mM, 0.5 mM FFA and 1% BSA for 24 h. The control cells were treated only with 1% BSA. (E) Cellular viability after incubating with different FFAs for various durations. Chang liver cells were incubated with 2 mM, 1 mM, and 0.5 mM FFA for 12 h, 24 h and 48 h respectively. The control cells were only treated with 1% BSA. Single points included the means of at least three independent experiments. (F) Effects of different fatty acids on triglyceride accumulation. The ratio of triglyceride accumulation to the total protein amount (TG/protein, mg/g) was evaluated as the concentration of TG in cell lysate after radio immunoprecipitation. The 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

f1-bt-23-400: The lipid accumulation. (A, B, C, D) Representative microscopy photograph. Chang liver cells were exposed to 2 mM, 1 mM, 0.5 mM FFA and 1% BSA for 24 h. The control cells were treated only with 1% BSA. (E) Cellular viability after incubating with different FFAs for various durations. Chang liver cells were incubated with 2 mM, 1 mM, and 0.5 mM FFA for 12 h, 24 h and 48 h respectively. The control cells were only treated with 1% BSA. Single points included the means of at least three independent experiments. (F) Effects of different fatty acids on triglyceride accumulation. The ratio of triglyceride accumulation to the total protein amount (TG/protein, mg/g) was evaluated as the concentration of TG in cell lysate after radio immunoprecipitation. The column represented the mean ± SD of three independent experiments. *p<0.05 versus control.
Mentions: After the Chang liver cells were exposed to successively graded concentrations of FFA (2 mM, 1 mM, 0.5 mM) for 24 h, oil red O staining was performed and the results shown in Fig. 1 A∼D indicating the lipid accumulation was FFA concentration-dependent. The cellular viability was tested using the methyl thiazolyl tetrazolium (MTT) method and the ratio of triglyceride (TG) accumulation to the total protein amount (TG/protein, mg/g) was assessed to confirm lipid accumulation. As shown in Fig. 1E, 1F, the cellular viability and TG/protein ratio exhibited a time-dependent and FFA concentration-dependent phenomenon. For overall considerations of the cellular viability and the lipid accumulation, the optimal incubation time was 24 hours and the optimal incubation concentration of FFA was 1 mM.

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