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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

IncRNA-HIF2PUT and HIF-2α expression levels were analyzed by real-time PCR in CRC tissue samples and in eight cell lines.Notes: (A) The expression levels of lncRNA-HIF2PUT and (B) HIF-2α were calculated as the ratio of targets in cancerous tissue/targets in adjacent normal tissue (R [C/N]). (C) The expression of lncRNA-HIF2PUT was significantly correlated with that of HIF-2α (P<0.0001). (D) The expression of lncRNA-HIF2PUT was coincident with that of HIF-2α in the eight cell lines (P<0.0001). (E and F) The expression of both lncRNA-HIF2PUT and HIF-2α were significantly higher in each of the seven CRC cell lines than in the normal colon epithelial cell line FHC. The expression of lncRNA-HIF2PUT and HIF-2α was highest in DLD-1 and HT29 cells. *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; CRC, colorectal cancer; PCR, polymerase chain reaction.
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f1-ott-8-3467: IncRNA-HIF2PUT and HIF-2α expression levels were analyzed by real-time PCR in CRC tissue samples and in eight cell lines.Notes: (A) The expression levels of lncRNA-HIF2PUT and (B) HIF-2α were calculated as the ratio of targets in cancerous tissue/targets in adjacent normal tissue (R [C/N]). (C) The expression of lncRNA-HIF2PUT was significantly correlated with that of HIF-2α (P<0.0001). (D) The expression of lncRNA-HIF2PUT was coincident with that of HIF-2α in the eight cell lines (P<0.0001). (E and F) The expression of both lncRNA-HIF2PUT and HIF-2α were significantly higher in each of the seven CRC cell lines than in the normal colon epithelial cell line FHC. The expression of lncRNA-HIF2PUT and HIF-2α was highest in DLD-1 and HT29 cells. *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; CRC, colorectal cancer; PCR, polymerase chain reaction.

Mentions: LncRNA-HIF2PUT and HIF-2α expression levels were assessed in a group of 20 patients with CRC. For each patient, lncRNA was isolated from cancerous tissues and adjacent nontumorous CRC tissues. The results showed that the expression of lncRNA-HIF2PUT was significantly correlated with that of HIF-2α in CRC tissue samples (R=0.5793, P<0.0001; Figure 1A–C). LncRNA-HIF2PUT and HIF-2α expression were also investigated in a normal colon epithelial cell line FHC and seven different CRC cell lines. The results revealed that the expression of lncRNA-HIF2PUT was coincident with that of HIF-2α in all eight cell lines (R=0.983, P<0.0001; Figure 1D, E). Moreover, expression of both lncRNA-HIF2PUT and HIF-2α were significantly higher in CRC cell lines than in FHC (Figure 1E). Also, the expression of both lncRNA-HIF2PUT and HIF-2α was higher in DLD-1 and HT29 cells than the other cell lines (Figure 1E). Therefore, we chose DLD-1 and HT29 as our candidate cell lines for a lncRNA-HIF2PUT knockdown experiment.


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)

IncRNA-HIF2PUT and HIF-2α expression levels were analyzed by real-time PCR in CRC tissue samples and in eight cell lines.Notes: (A) The expression levels of lncRNA-HIF2PUT and (B) HIF-2α were calculated as the ratio of targets in cancerous tissue/targets in adjacent normal tissue (R [C/N]). (C) The expression of lncRNA-HIF2PUT was significantly correlated with that of HIF-2α (P<0.0001). (D) The expression of lncRNA-HIF2PUT was coincident with that of HIF-2α in the eight cell lines (P<0.0001). (E and F) The expression of both lncRNA-HIF2PUT and HIF-2α were significantly higher in each of the seven CRC cell lines than in the normal colon epithelial cell line FHC. The expression of lncRNA-HIF2PUT and HIF-2α was highest in DLD-1 and HT29 cells. *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; CRC, colorectal cancer; PCR, polymerase chain reaction.
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Related In: Results  -  Collection

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f1-ott-8-3467: IncRNA-HIF2PUT and HIF-2α expression levels were analyzed by real-time PCR in CRC tissue samples and in eight cell lines.Notes: (A) The expression levels of lncRNA-HIF2PUT and (B) HIF-2α were calculated as the ratio of targets in cancerous tissue/targets in adjacent normal tissue (R [C/N]). (C) The expression of lncRNA-HIF2PUT was significantly correlated with that of HIF-2α (P<0.0001). (D) The expression of lncRNA-HIF2PUT was coincident with that of HIF-2α in the eight cell lines (P<0.0001). (E and F) The expression of both lncRNA-HIF2PUT and HIF-2α were significantly higher in each of the seven CRC cell lines than in the normal colon epithelial cell line FHC. The expression of lncRNA-HIF2PUT and HIF-2α was highest in DLD-1 and HT29 cells. *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; CRC, colorectal cancer; PCR, polymerase chain reaction.
Mentions: LncRNA-HIF2PUT and HIF-2α expression levels were assessed in a group of 20 patients with CRC. For each patient, lncRNA was isolated from cancerous tissues and adjacent nontumorous CRC tissues. The results showed that the expression of lncRNA-HIF2PUT was significantly correlated with that of HIF-2α in CRC tissue samples (R=0.5793, P<0.0001; Figure 1A–C). LncRNA-HIF2PUT and HIF-2α expression were also investigated in a normal colon epithelial cell line FHC and seven different CRC cell lines. The results revealed that the expression of lncRNA-HIF2PUT was coincident with that of HIF-2α in all eight cell lines (R=0.983, P<0.0001; Figure 1D, E). Moreover, expression of both lncRNA-HIF2PUT and HIF-2α were significantly higher in CRC cell lines than in FHC (Figure 1E). Also, the expression of both lncRNA-HIF2PUT and HIF-2α was higher in DLD-1 and HT29 cells than the other cell lines (Figure 1E). Therefore, we chose DLD-1 and HT29 as our candidate cell lines for a lncRNA-HIF2PUT knockdown experiment.

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