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Knockdown of a HIF-2α promoter upstream long noncoding RNA impairs colorectal cancer stem cell properties in vitro through HIF-2α downregulation.

Yao J, Li J, Geng P, Li Y, Chen H, Zhu Y - Onco Targets Ther (2015)

Bottom Line: In this study, we found an lncRNA that is a promoter upstream transcript of hypoxia-inducible factor-2α (HIF-2α), and we named it "lncRNA-HIF2PUT".Herein, we showed that the expression of lncRNA-HIF2PUT was significantly correlated with HIF-2α in colorectal cancer (CRC) tissues.LncRNA-HIF2PUTsmall interfering RNA transfection resulted in decreased stemness genes expression, impaired colony formation, and spheroid formation ability, retarded migration, and invasion of the cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Oncology, People's Liberation Army No 161 Hospital, Wuhan, People's Republic of China.

ABSTRACT
Currently, various long noncoding RNAs (lncRNAs) have been identified as key regulators of multiple cancers. However, cancer stem cell (CSC)-related lncRNAs have rarely been reported. In this study, we found an lncRNA that is a promoter upstream transcript of hypoxia-inducible factor-2α (HIF-2α), and we named it "lncRNA-HIF2PUT". The function of HIF-2α is closely connected with "stem cell-like" properties, and the function of PROMPTs is often associated with the adjacent protein-coding transcripts. Herein, we showed that the expression of lncRNA-HIF2PUT was significantly correlated with HIF-2α in colorectal cancer (CRC) tissues. Knockdown of lncRNA-HIF2PUT blocked the HIF-2α expression and inhibited the CSC properties in CRC cell lines DLD-1 and HT29. LncRNA-HIF2PUTsmall interfering RNA transfection resulted in decreased stemness genes expression, impaired colony formation, and spheroid formation ability, retarded migration, and invasion of the cells. These data suggest that lncRNA-HIF2PUT may be a regulator of HIF-2α and a mediator of CSCs in CRC.

No MeSH data available.


Related in: MedlinePlus

Knockdown of lncRNA-HIF2PUT inhibited the expression of HIF-2α.Notes: (A) Quantification analysis by real-time PCR showed that the expression levels of both lncRNA-HIF2PUT and HIF-2α in lncRNA-HIF2PUT siRNA groups were significantly knocked down in DLD-1 and HT29 cell lines. (B) Western blot showed that the HIF-2α protein expression was also markedly decreased. *P<0.05.Abbreviations: lncRNA-HIF2PUT, long noncoding RNA that is a promoter upstream transcript (PROMPT) of hypoxia-inducible factor-2α; HIF-2α, hypoxia-inducible factor-2 alpha; PCR, polymerase chain reaction; siRNA, small interfering RNA; NC, noncancerous.
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f2-ott-8-3467: Knockdown of lncRNA-HIF2PUT inhibited the expression of HIF-2α.Notes: (A) Quantification analysis by real-time PCR showed that the expression levels of both lncRNA-HIF2PUT and HIF-2α in lncRNA-HIF2PUT siRNA groups were significantly knocked down in DLD-1 and HT29 cell lines. (B) Western blot showed that the HIF-2α protein expression was also markedly decreased. *P<0.05.Abbreviations: lncRNA-HIF2PUT, long noncoding RNA that is a promoter upstream transcript (PROMPT) of hypoxia-inducible factor-2α; HIF-2α, hypoxia-inducible factor-2 alpha; PCR, polymerase chain reaction; siRNA, small interfering RNA; NC, noncancerous.

Mentions: LncRNA-HIF2PUT siRNA was transfected into DLD-1 and HT29. Quantification analysis showed that lncRNA-HIF2PUT expression level in the lncRNA-HIF2PUT siRNA group was significantly knocked down in DLD-1 and HT29 cells (Figure 2A). Using RT-qPCR and Western blots, we showed that HIF-2α expression was also markedly decreased in both mRNA level and protein level (Figure 2A, B).


Knockdown of a HIF-2α promoter upstream long noncoding RNA impairs colorectal cancer stem cell properties in vitro through HIF-2α downregulation.

Yao J, Li J, Geng P, Li Y, Chen H, Zhu Y - Onco Targets Ther (2015)

Knockdown of lncRNA-HIF2PUT inhibited the expression of HIF-2α.Notes: (A) Quantification analysis by real-time PCR showed that the expression levels of both lncRNA-HIF2PUT and HIF-2α in lncRNA-HIF2PUT siRNA groups were significantly knocked down in DLD-1 and HT29 cell lines. (B) Western blot showed that the HIF-2α protein expression was also markedly decreased. *P<0.05.Abbreviations: lncRNA-HIF2PUT, long noncoding RNA that is a promoter upstream transcript (PROMPT) of hypoxia-inducible factor-2α; HIF-2α, hypoxia-inducible factor-2 alpha; PCR, polymerase chain reaction; siRNA, small interfering RNA; NC, noncancerous.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4664519&req=5

f2-ott-8-3467: Knockdown of lncRNA-HIF2PUT inhibited the expression of HIF-2α.Notes: (A) Quantification analysis by real-time PCR showed that the expression levels of both lncRNA-HIF2PUT and HIF-2α in lncRNA-HIF2PUT siRNA groups were significantly knocked down in DLD-1 and HT29 cell lines. (B) Western blot showed that the HIF-2α protein expression was also markedly decreased. *P<0.05.Abbreviations: lncRNA-HIF2PUT, long noncoding RNA that is a promoter upstream transcript (PROMPT) of hypoxia-inducible factor-2α; HIF-2α, hypoxia-inducible factor-2 alpha; PCR, polymerase chain reaction; siRNA, small interfering RNA; NC, noncancerous.
Mentions: LncRNA-HIF2PUT siRNA was transfected into DLD-1 and HT29. Quantification analysis showed that lncRNA-HIF2PUT expression level in the lncRNA-HIF2PUT siRNA group was significantly knocked down in DLD-1 and HT29 cells (Figure 2A). Using RT-qPCR and Western blots, we showed that HIF-2α expression was also markedly decreased in both mRNA level and protein level (Figure 2A, B).

Bottom Line: In this study, we found an lncRNA that is a promoter upstream transcript of hypoxia-inducible factor-2α (HIF-2α), and we named it "lncRNA-HIF2PUT".Herein, we showed that the expression of lncRNA-HIF2PUT was significantly correlated with HIF-2α in colorectal cancer (CRC) tissues.LncRNA-HIF2PUTsmall interfering RNA transfection resulted in decreased stemness genes expression, impaired colony formation, and spheroid formation ability, retarded migration, and invasion of the cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Oncology, People's Liberation Army No 161 Hospital, Wuhan, People's Republic of China.

ABSTRACT
Currently, various long noncoding RNAs (lncRNAs) have been identified as key regulators of multiple cancers. However, cancer stem cell (CSC)-related lncRNAs have rarely been reported. In this study, we found an lncRNA that is a promoter upstream transcript of hypoxia-inducible factor-2α (HIF-2α), and we named it "lncRNA-HIF2PUT". The function of HIF-2α is closely connected with "stem cell-like" properties, and the function of PROMPTs is often associated with the adjacent protein-coding transcripts. Herein, we showed that the expression of lncRNA-HIF2PUT was significantly correlated with HIF-2α in colorectal cancer (CRC) tissues. Knockdown of lncRNA-HIF2PUT blocked the HIF-2α expression and inhibited the CSC properties in CRC cell lines DLD-1 and HT29. LncRNA-HIF2PUTsmall interfering RNA transfection resulted in decreased stemness genes expression, impaired colony formation, and spheroid formation ability, retarded migration, and invasion of the cells. These data suggest that lncRNA-HIF2PUT may be a regulator of HIF-2α and a mediator of CSCs in CRC.

No MeSH data available.


Related in: MedlinePlus