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Long non-coding RNAs expression profile in HepG2 cells reveals the potential role of long non-coding RNAs in the cholesterol metabolism.

Liu G, Zheng X, Xu Y, Lu J, Chen J, Huang X - Chin. Med. J. (2015)

Bottom Line: Our aim was to identify important lncRNAs that might play an important role in contributing to the benefits of epigallocatechin-3-gallate (EGCG) on cholesterol metabolism.Bioinformatic analysis found five matched lncRNA-mRNA pairs for five differentially expressed lncRNAs and four differentially expressed mRNA.After AT102202 knockdown in HepG2, we observed that the level of HMGCR expression was significantly increased relative to the scrambled small interfering RNA control (P < 0.05).

View Article: PubMed Central - PubMed

Affiliation: Department of Special Medical Treatment Center, Fuwai Hospital, National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100037, China.

ABSTRACT

Background: Green tea has been shown to improve cholesterol metabolism in animal studies, but the molecular mechanisms underlying this function have not been fully understood. Long non-coding RNAs (lncRNAs) have recently emerged as a major class of regulatory molecules involved in a broad range of biological processes and complex diseases. Our aim was to identify important lncRNAs that might play an important role in contributing to the benefits of epigallocatechin-3-gallate (EGCG) on cholesterol metabolism.

Methods: Microarrays was used to reveal the lncRNA and mRNA profiles in green tea polyphenol(-)-epigallocatechin gallate in cultured human liver (HepG2) hepatocytes treated with EGCG and bioinformatic analyses of the predicted target genes were performed to identify lncRNA-mRNA targeting relationships. RNA interference was used to investigate the role of lncRNAs in cholesterol metabolism.

Results: The expression levels of 15 genes related to cholesterol metabolism and 285 lncRNAs were changed by EGCG treatment. Bioinformatic analysis found five matched lncRNA-mRNA pairs for five differentially expressed lncRNAs and four differentially expressed mRNA. In particular, the lncRNA AT102202 and its potential targets mRNA-3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGCR) were identified. Using a real-time polymerase chain reaction technique, we confirmed that EGCG down-regulated mRNA expression level of the HMGCR and up-regulated expression of AT102202. After AT102202 knockdown in HepG2, we observed that the level of HMGCR expression was significantly increased relative to the scrambled small interfering RNA control (P < 0.05).

Conclusions: Our results indicated that EGCG improved cholesterol metabolism and meanwhile changed the lncRNAs expression profile in HepG2 cells. LncRNAs may play an important role in the cholesterol metabolism.

Show MeSH
The expression of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) mRNA expression following AT102202 knockdown (siRNA124 at 18 nmol/L) in HepG2 cells with EGCG (10 or 25 μmol/L) treatment for 24 hours. The level of HMGCR expression was measured using quantitative real-time PCR and error bars indicate the standard error of the mean for 6 technical replicates and expression values are normalized to scramble siRNA controls.
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Figure 3: The expression of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) mRNA expression following AT102202 knockdown (siRNA124 at 18 nmol/L) in HepG2 cells with EGCG (10 or 25 μmol/L) treatment for 24 hours. The level of HMGCR expression was measured using quantitative real-time PCR and error bars indicate the standard error of the mean for 6 technical replicates and expression values are normalized to scramble siRNA controls.

Mentions: Given the correlated expression of AT102202 and HMGCR, we next aimed to determine the effect of AT102202 knockdown on HMGCR expression in HepG2 cells treated with or without EGCG. Using qRT-PCR, we determined the expression of HMGCR following siRNA-mediated knockdown of AT102202. As a result, we found that the level of HMGCR expression was significantly increased following AT102202 knockdown relative to the scrambled siRNA control [Figure 3]. These results suggest that AT102202 regulates HMGCR expression, and EGCG inhibits the HMGCR expression partially through by AT102202.


Long non-coding RNAs expression profile in HepG2 cells reveals the potential role of long non-coding RNAs in the cholesterol metabolism.

Liu G, Zheng X, Xu Y, Lu J, Chen J, Huang X - Chin. Med. J. (2015)

The expression of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) mRNA expression following AT102202 knockdown (siRNA124 at 18 nmol/L) in HepG2 cells with EGCG (10 or 25 μmol/L) treatment for 24 hours. The level of HMGCR expression was measured using quantitative real-time PCR and error bars indicate the standard error of the mean for 6 technical replicates and expression values are normalized to scramble siRNA controls.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4837827&req=5

Figure 3: The expression of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) mRNA expression following AT102202 knockdown (siRNA124 at 18 nmol/L) in HepG2 cells with EGCG (10 or 25 μmol/L) treatment for 24 hours. The level of HMGCR expression was measured using quantitative real-time PCR and error bars indicate the standard error of the mean for 6 technical replicates and expression values are normalized to scramble siRNA controls.
Mentions: Given the correlated expression of AT102202 and HMGCR, we next aimed to determine the effect of AT102202 knockdown on HMGCR expression in HepG2 cells treated with or without EGCG. Using qRT-PCR, we determined the expression of HMGCR following siRNA-mediated knockdown of AT102202. As a result, we found that the level of HMGCR expression was significantly increased following AT102202 knockdown relative to the scrambled siRNA control [Figure 3]. These results suggest that AT102202 regulates HMGCR expression, and EGCG inhibits the HMGCR expression partially through by AT102202.

Bottom Line: Our aim was to identify important lncRNAs that might play an important role in contributing to the benefits of epigallocatechin-3-gallate (EGCG) on cholesterol metabolism.Bioinformatic analysis found five matched lncRNA-mRNA pairs for five differentially expressed lncRNAs and four differentially expressed mRNA.After AT102202 knockdown in HepG2, we observed that the level of HMGCR expression was significantly increased relative to the scrambled small interfering RNA control (P < 0.05).

View Article: PubMed Central - PubMed

Affiliation: Department of Special Medical Treatment Center, Fuwai Hospital, National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100037, China.

ABSTRACT

Background: Green tea has been shown to improve cholesterol metabolism in animal studies, but the molecular mechanisms underlying this function have not been fully understood. Long non-coding RNAs (lncRNAs) have recently emerged as a major class of regulatory molecules involved in a broad range of biological processes and complex diseases. Our aim was to identify important lncRNAs that might play an important role in contributing to the benefits of epigallocatechin-3-gallate (EGCG) on cholesterol metabolism.

Methods: Microarrays was used to reveal the lncRNA and mRNA profiles in green tea polyphenol(-)-epigallocatechin gallate in cultured human liver (HepG2) hepatocytes treated with EGCG and bioinformatic analyses of the predicted target genes were performed to identify lncRNA-mRNA targeting relationships. RNA interference was used to investigate the role of lncRNAs in cholesterol metabolism.

Results: The expression levels of 15 genes related to cholesterol metabolism and 285 lncRNAs were changed by EGCG treatment. Bioinformatic analysis found five matched lncRNA-mRNA pairs for five differentially expressed lncRNAs and four differentially expressed mRNA. In particular, the lncRNA AT102202 and its potential targets mRNA-3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGCR) were identified. Using a real-time polymerase chain reaction technique, we confirmed that EGCG down-regulated mRNA expression level of the HMGCR and up-regulated expression of AT102202. After AT102202 knockdown in HepG2, we observed that the level of HMGCR expression was significantly increased relative to the scrambled small interfering RNA control (P < 0.05).

Conclusions: Our results indicated that EGCG improved cholesterol metabolism and meanwhile changed the lncRNAs expression profile in HepG2 cells. LncRNAs may play an important role in the cholesterol metabolism.

Show MeSH