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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
Effects of the addition of epigallocatechin gallate (EGCG) on the level of AT102202 (a), 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) (b), and LDL receptor (LDLR) mRNA expression (c). Total RNA were harvested from HepG2 cells treated with vehicle control, 10 or 25 μmol/L-EGCG for 24 hours and the level mRNA expression was measured using quantitative real-time PCR and normalised to the mRNA expression level of the GADPH gene. The levels of the vehicle-control-treated group are set at 100%, and the levels are presented as fold inductions relative to the vehicle-control-treated group. Values are means, with their standard error (n=6). *P <0.05 vs. control group.
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Figure 1: Effects of the addition of epigallocatechin gallate (EGCG) on the level of AT102202 (a), 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) (b), and LDL receptor (LDLR) mRNA expression (c). Total RNA were harvested from HepG2 cells treated with vehicle control, 10 or 25 μmol/L-EGCG for 24 hours and the level mRNA expression was measured using quantitative real-time PCR and normalised to the mRNA expression level of the GADPH gene. The levels of the vehicle-control-treated group are set at 100%, and the levels are presented as fold inductions relative to the vehicle-control-treated group. Values are means, with their standard error (n=6). *P <0.05 vs. control group.

Mentions: In the present study, we focused on investigating the effect of EGCG on HMGCR, AT102202 and LDL receptor expression, and qRT-PCR was carried out to confirm the effect of EGCG on expression levels of these genes. As expected, the addition of 10 and 25 μM of EGCG significantly increased the level of expression of AT102202 and LDL receptor, meanwhile decreased HMGCR expression [Figure 1]. Furthermore, we confirmed that the expression of AT102202 and its predicted target gene-HMGCR was linked.


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)

Effects of the addition of epigallocatechin gallate (EGCG) on the level of AT102202 (a), 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) (b), and LDL receptor (LDLR) mRNA expression (c). Total RNA were harvested from HepG2 cells treated with vehicle control, 10 or 25 μmol/L-EGCG for 24 hours and the level mRNA expression was measured using quantitative real-time PCR and normalised to the mRNA expression level of the GADPH gene. The levels of the vehicle-control-treated group are set at 100%, and the levels are presented as fold inductions relative to the vehicle-control-treated group. Values are means, with their standard error (n=6). *P <0.05 vs. control group.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 1: Effects of the addition of epigallocatechin gallate (EGCG) on the level of AT102202 (a), 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) (b), and LDL receptor (LDLR) mRNA expression (c). Total RNA were harvested from HepG2 cells treated with vehicle control, 10 or 25 μmol/L-EGCG for 24 hours and the level mRNA expression was measured using quantitative real-time PCR and normalised to the mRNA expression level of the GADPH gene. The levels of the vehicle-control-treated group are set at 100%, and the levels are presented as fold inductions relative to the vehicle-control-treated group. Values are means, with their standard error (n=6). *P <0.05 vs. control group.
Mentions: In the present study, we focused on investigating the effect of EGCG on HMGCR, AT102202 and LDL receptor expression, and qRT-PCR was carried out to confirm the effect of EGCG on expression levels of these genes. As expected, the addition of 10 and 25 μM of EGCG significantly increased the level of expression of AT102202 and LDL receptor, meanwhile decreased HMGCR expression [Figure 1]. Furthermore, we confirmed that the expression of AT102202 and its predicted target gene-HMGCR was linked.

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