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Long noncoding RNA CCAT2 promotes breast tumor growth by regulating the Wnt signaling pathway.

Cai Y, He J, Zhang D - Onco Targets Ther (2015)

Bottom Line: In addition to protein-coding genes, the human genome makes a large amount of noncoding RNAs, including microRNAs and long noncoding RNAs (lncRNAs).Also, the biological function of CCAT2 was explored and the results showed silencing of CCAT2 could suppress cell growth in vitro and tumor formation in vivo.Finally, our results revealed that the abnormal expression of CCAT2 could influence the Wnt signaling pathway.

View Article: PubMed Central - PubMed

Affiliation: Department of Geriatric Oncology, The General Hospital of Chinese People's Liberation Army, Beijing City, People's Republic of China.

ABSTRACT
In addition to protein-coding genes, the human genome makes a large amount of noncoding RNAs, including microRNAs and long noncoding RNAs (lncRNAs). Emerging evidence indicates that lncRNAs could have a critical role in the regulation of cellular processes such as cell growth and apoptosis as well as cancer progression and metastasis. The lncRNA CCAT2 is dysregulated in several cancers such as colon cancer, non-small cell lung cancer, esophageal squamous cell carcinoma, gastric cancer, and breast cancer; however, the contributions of CCAT2 to breast cancer remain largely unknown. In the current paper, we first confirmed the high expression level of CCAT2 in breast cancer tissues and breast cancer cell lines by reverse transcription quantitative polymerase chain reaction (RT-qPCR) assay, and we further analyzed the relationship between CCAT2 expression and clinical prognostic factors. Also, the biological function of CCAT2 was explored and the results showed silencing of CCAT2 could suppress cell growth in vitro and tumor formation in vivo. Finally, our results revealed that the abnormal expression of CCAT2 could influence the Wnt signaling pathway. In conclusion, lncRNA CCAT2 might be considered as a novel molecule involved in breast cancer development, which provides a potential therapeutic target for breast cancer.

No MeSH data available.


Related in: MedlinePlus

Suppressing CCAT2 expression affects the Wnt/β-catenin signaling pathway.Notes: (A) A Western blot assay was performed to detect the β-catenin content in the nucleus and cytoplasm of MCF-7 and MDA-MB-231 cells treated with CCAT2 siRNA or Wnt signaling inhibitor FH 535 (10 μM) for 48 hours. The data showed that si-CCAT2 inhibited the β-catenin expression at transcriptional level. Histone 3 was used as an internal control for nuclear protein and actin was used as the control for cytoplasm protein. (B) The CCND1 and c-myc expression levels were detected by a RT-qPCR assay. The MCF-7 and MDA-MB-231 cells were treated with si-CCAT2 or 10 μM FH 535, alone or combined, for 48 hours. GAPDH was used as an internal control. (C) TOP/FOP Flash detection system was used to investigate the effects of si-CCAT2 or FH 535, alone or combined, on Wnt/β-catenin in MCF-7 and MD-MB-231 cells. The data showed si-CCAT2 inhibited the β-catenin transcriptional activity. The experiments were all repeated at least three times. *P<0.05, and #P<0.001 vs the control.Abbreviations: RT-qPCR, reverse transcription quantitative polymerase chain reaction; si-CCAT2, siRNA specifically targeting CCAT2; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; si-NS, nonspecific siRNA.
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f5-ott-8-2657: Suppressing CCAT2 expression affects the Wnt/β-catenin signaling pathway.Notes: (A) A Western blot assay was performed to detect the β-catenin content in the nucleus and cytoplasm of MCF-7 and MDA-MB-231 cells treated with CCAT2 siRNA or Wnt signaling inhibitor FH 535 (10 μM) for 48 hours. The data showed that si-CCAT2 inhibited the β-catenin expression at transcriptional level. Histone 3 was used as an internal control for nuclear protein and actin was used as the control for cytoplasm protein. (B) The CCND1 and c-myc expression levels were detected by a RT-qPCR assay. The MCF-7 and MDA-MB-231 cells were treated with si-CCAT2 or 10 μM FH 535, alone or combined, for 48 hours. GAPDH was used as an internal control. (C) TOP/FOP Flash detection system was used to investigate the effects of si-CCAT2 or FH 535, alone or combined, on Wnt/β-catenin in MCF-7 and MD-MB-231 cells. The data showed si-CCAT2 inhibited the β-catenin transcriptional activity. The experiments were all repeated at least three times. *P<0.05, and #P<0.001 vs the control.Abbreviations: RT-qPCR, reverse transcription quantitative polymerase chain reaction; si-CCAT2, siRNA specifically targeting CCAT2; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; si-NS, nonspecific siRNA.

Mentions: To better understand the detailed regulation mechanism of CCAT2 in breast cancer, we first examined whether suppressing CCAT2 affects the Wnt signaling pathway, whose activation plays an important role in breast cancer development. Using the si-CCAT2 group and the si-NS group of the MCF-7 and MDA-MB-231 cells, as well as the Wnt signaling inhibitor FH 535, we found that suppressing the expression of CCAT2 decreased the levels of β-catenin both in the cytoplasm and nucleus via Western blot (Figure 5A), but FH 535 only affected the β-catenin recruitment, and the combination of si-CCAT2 and FH 535 could induce a synergetic effect on Wnt signaling activity. Moreover, CCAT2 knockdown reduced the expression of CCND1 and c-myc (classic downstream genes of the Wnt/β-catenin signaling pathway) via RT-qPCR in both MCF-7 and MDA-MB-231 cells (Figure 5B). In addition, the TOP/FOP Flash luciferase reporter system was employed in MCF-7 and MDA-MB-231 cells; the result showed that suppressing CCAT2 expression inhibited the Wnt/β-catenin signaling pathway transcriptional activity, and combination of siCCAT2 and FH 535 could synergistically inhibit the Wnt signaling (Figure 5C). Thus, these results suggest that suppressing CCAT2 expression affects the Wnt/β-catenin signaling pathway.


Long noncoding RNA CCAT2 promotes breast tumor growth by regulating the Wnt signaling pathway.

Cai Y, He J, Zhang D - Onco Targets Ther (2015)

Suppressing CCAT2 expression affects the Wnt/β-catenin signaling pathway.Notes: (A) A Western blot assay was performed to detect the β-catenin content in the nucleus and cytoplasm of MCF-7 and MDA-MB-231 cells treated with CCAT2 siRNA or Wnt signaling inhibitor FH 535 (10 μM) for 48 hours. The data showed that si-CCAT2 inhibited the β-catenin expression at transcriptional level. Histone 3 was used as an internal control for nuclear protein and actin was used as the control for cytoplasm protein. (B) The CCND1 and c-myc expression levels were detected by a RT-qPCR assay. The MCF-7 and MDA-MB-231 cells were treated with si-CCAT2 or 10 μM FH 535, alone or combined, for 48 hours. GAPDH was used as an internal control. (C) TOP/FOP Flash detection system was used to investigate the effects of si-CCAT2 or FH 535, alone or combined, on Wnt/β-catenin in MCF-7 and MD-MB-231 cells. The data showed si-CCAT2 inhibited the β-catenin transcriptional activity. The experiments were all repeated at least three times. *P<0.05, and #P<0.001 vs the control.Abbreviations: RT-qPCR, reverse transcription quantitative polymerase chain reaction; si-CCAT2, siRNA specifically targeting CCAT2; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; si-NS, nonspecific siRNA.
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Related In: Results  -  Collection

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f5-ott-8-2657: Suppressing CCAT2 expression affects the Wnt/β-catenin signaling pathway.Notes: (A) A Western blot assay was performed to detect the β-catenin content in the nucleus and cytoplasm of MCF-7 and MDA-MB-231 cells treated with CCAT2 siRNA or Wnt signaling inhibitor FH 535 (10 μM) for 48 hours. The data showed that si-CCAT2 inhibited the β-catenin expression at transcriptional level. Histone 3 was used as an internal control for nuclear protein and actin was used as the control for cytoplasm protein. (B) The CCND1 and c-myc expression levels were detected by a RT-qPCR assay. The MCF-7 and MDA-MB-231 cells were treated with si-CCAT2 or 10 μM FH 535, alone or combined, for 48 hours. GAPDH was used as an internal control. (C) TOP/FOP Flash detection system was used to investigate the effects of si-CCAT2 or FH 535, alone or combined, on Wnt/β-catenin in MCF-7 and MD-MB-231 cells. The data showed si-CCAT2 inhibited the β-catenin transcriptional activity. The experiments were all repeated at least three times. *P<0.05, and #P<0.001 vs the control.Abbreviations: RT-qPCR, reverse transcription quantitative polymerase chain reaction; si-CCAT2, siRNA specifically targeting CCAT2; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; si-NS, nonspecific siRNA.
Mentions: To better understand the detailed regulation mechanism of CCAT2 in breast cancer, we first examined whether suppressing CCAT2 affects the Wnt signaling pathway, whose activation plays an important role in breast cancer development. Using the si-CCAT2 group and the si-NS group of the MCF-7 and MDA-MB-231 cells, as well as the Wnt signaling inhibitor FH 535, we found that suppressing the expression of CCAT2 decreased the levels of β-catenin both in the cytoplasm and nucleus via Western blot (Figure 5A), but FH 535 only affected the β-catenin recruitment, and the combination of si-CCAT2 and FH 535 could induce a synergetic effect on Wnt signaling activity. Moreover, CCAT2 knockdown reduced the expression of CCND1 and c-myc (classic downstream genes of the Wnt/β-catenin signaling pathway) via RT-qPCR in both MCF-7 and MDA-MB-231 cells (Figure 5B). In addition, the TOP/FOP Flash luciferase reporter system was employed in MCF-7 and MDA-MB-231 cells; the result showed that suppressing CCAT2 expression inhibited the Wnt/β-catenin signaling pathway transcriptional activity, and combination of siCCAT2 and FH 535 could synergistically inhibit the Wnt signaling (Figure 5C). Thus, these results suggest that suppressing CCAT2 expression affects the Wnt/β-catenin signaling pathway.

Bottom Line: In addition to protein-coding genes, the human genome makes a large amount of noncoding RNAs, including microRNAs and long noncoding RNAs (lncRNAs).Also, the biological function of CCAT2 was explored and the results showed silencing of CCAT2 could suppress cell growth in vitro and tumor formation in vivo.Finally, our results revealed that the abnormal expression of CCAT2 could influence the Wnt signaling pathway.

View Article: PubMed Central - PubMed

Affiliation: Department of Geriatric Oncology, The General Hospital of Chinese People's Liberation Army, Beijing City, People's Republic of China.

ABSTRACT
In addition to protein-coding genes, the human genome makes a large amount of noncoding RNAs, including microRNAs and long noncoding RNAs (lncRNAs). Emerging evidence indicates that lncRNAs could have a critical role in the regulation of cellular processes such as cell growth and apoptosis as well as cancer progression and metastasis. The lncRNA CCAT2 is dysregulated in several cancers such as colon cancer, non-small cell lung cancer, esophageal squamous cell carcinoma, gastric cancer, and breast cancer; however, the contributions of CCAT2 to breast cancer remain largely unknown. In the current paper, we first confirmed the high expression level of CCAT2 in breast cancer tissues and breast cancer cell lines by reverse transcription quantitative polymerase chain reaction (RT-qPCR) assay, and we further analyzed the relationship between CCAT2 expression and clinical prognostic factors. Also, the biological function of CCAT2 was explored and the results showed silencing of CCAT2 could suppress cell growth in vitro and tumor formation in vivo. Finally, our results revealed that the abnormal expression of CCAT2 could influence the Wnt signaling pathway. In conclusion, lncRNA CCAT2 might be considered as a novel molecule involved in breast cancer development, which provides a potential therapeutic target for breast cancer.

No MeSH data available.


Related in: MedlinePlus