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MicroRNA-1269 promotes proliferation in human hepatocellular carcinoma via downregulation of FOXO1.

Yang XW, Shen GZ, Cao LQ, Jiang XF, Peng HP, Shen G, Chen D, Xue P - BMC Cancer (2014)

Bottom Line: Furthermore, we demonstrated that FOXO1 was a direct target of miR-1269.Suppression of FOXO1 by miR-1269 was associated with dysregulation of p21, cyclin D1, phosphorylated Rb and Ki67 expression, thereby playing an essential role in the growth of HCC cells.Our study indicated that overexpression of miR-1269 promotes cell proliferation in HCC through directly suppressing FOXO1, and functions as an oncomiR in HCC.

View Article: PubMed Central - PubMed

Affiliation: Department of Hepatobiliary Surgery, the Second Affiliated Hospital of Guangzhou, Medical University, Guangzhou 510260, China. drxueping@medmail.com.cn.

ABSTRACT

Background: Hepatocellular carcinoma (HCC) is one of the most common malignancies and a major cause of cancer-related mortality in the world. MicroRNAs (miRNAs) are small, noncoding RNAs that play essential roles in various stages during cancer progression. The aim of the current study was to elucidate the role of miR-1269 in the pathogenesis of HCC.

Methods: The expression of miR-1269 in HCC cells and tissues were determined by Real-time PCR analysis. Cell viability, colony formation and anchorage-independent growth ability assays were performed to examine cell proliferative capacity and tumorigenicity. Flow cytometry analysis was conducted to determine cell cycle progression. The expression of p21, CyclinD1, phosphorylated Rb, Rb and FOXO1 were examined by Western blotting analysis. Luciferase assay was used to determine whether FOXO1 is the direct target of miR-1269.

Results: miR-1269 was upregulated in HCC cells and tissues. Ectopic miR-1269 expression promoted, but inhibition of miR-1269 reduced, proliferation, tumorigenicity and cell cycle progression of HCC cells. Furthermore, we demonstrated that FOXO1 was a direct target of miR-1269. Suppression of FOXO1 by miR-1269 was associated with dysregulation of p21, cyclin D1, phosphorylated Rb and Ki67 expression, thereby playing an essential role in the growth of HCC cells.

Conclusions: Our study indicated that overexpression of miR-1269 promotes cell proliferation in HCC through directly suppressing FOXO1, and functions as an oncomiR in HCC.

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Related in: MedlinePlus

MiR-1269 enhances the proliferation and tumorigenicity of HCC cells. A. Real-time PCR analysis of miR-1269 in BEL-7404 and Huh7 cells stably overexpressing miR-1269. Transcript levels were normalized to U6 expression. B. The effects of miR-1269 overexpression on cell viability of the indicated cell lines analyzed by MTT assay. C. Representative micrographs (left panel) and quantification (right panel) of crystal violet stained colonies formed by the indicated cell lines. D. Representative micrographs (left panel) and quantification (right panel) of cell colonies determined by anchorage-independent growth ability assay. E. The effect of miR-1269 on cell cycle progression of the indicated cell lines analyzed by flow cytometry. Each bar represents the mean ± SD of three independent experiments. * P < 0.05.
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Fig2: MiR-1269 enhances the proliferation and tumorigenicity of HCC cells. A. Real-time PCR analysis of miR-1269 in BEL-7404 and Huh7 cells stably overexpressing miR-1269. Transcript levels were normalized to U6 expression. B. The effects of miR-1269 overexpression on cell viability of the indicated cell lines analyzed by MTT assay. C. Representative micrographs (left panel) and quantification (right panel) of crystal violet stained colonies formed by the indicated cell lines. D. Representative micrographs (left panel) and quantification (right panel) of cell colonies determined by anchorage-independent growth ability assay. E. The effect of miR-1269 on cell cycle progression of the indicated cell lines analyzed by flow cytometry. Each bar represents the mean ± SD of three independent experiments. * P < 0.05.

Mentions: To explore the biological function of miR-1269 in HCC progression, BEL-7404 and Huh7 cells stably overexpressing miR-1269 were established (Figure 2A). Cell viability was then measured by MTT assay. A shown in Figure 2B, ectopic expression of miR-1269 increased the growth rate of both HCC cell lines. Colony formation assay showed that upregulation of miR-1269 promoted the colony formation capacity of BEL-7404 and Huh7 cells (Figure 2C). Consistently, anchorage-independent growth assay revealed that the cells stably expressing miR-1269 formed more and larger-sized colonies than the control cells (Figure 2D). Furthermore, cell cycle analysis by flow cytometry showed a dramatic decrease in the percentage of cells in the G1/G0 phase and an increase in the percentage of cells in the S phase in miR-1269-overexpressing cells (Figure 2E). Collectively, these results suggest that upregulation of miR-1269 enhanced the proliferation, tumorigenicity and cell cycle progression of HCC cells.Figure 2


MicroRNA-1269 promotes proliferation in human hepatocellular carcinoma via downregulation of FOXO1.

Yang XW, Shen GZ, Cao LQ, Jiang XF, Peng HP, Shen G, Chen D, Xue P - BMC Cancer (2014)

MiR-1269 enhances the proliferation and tumorigenicity of HCC cells. A. Real-time PCR analysis of miR-1269 in BEL-7404 and Huh7 cells stably overexpressing miR-1269. Transcript levels were normalized to U6 expression. B. The effects of miR-1269 overexpression on cell viability of the indicated cell lines analyzed by MTT assay. C. Representative micrographs (left panel) and quantification (right panel) of crystal violet stained colonies formed by the indicated cell lines. D. Representative micrographs (left panel) and quantification (right panel) of cell colonies determined by anchorage-independent growth ability assay. E. The effect of miR-1269 on cell cycle progression of the indicated cell lines analyzed by flow cytometry. Each bar represents the mean ± SD of three independent experiments. * P < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4265494&req=5

Fig2: MiR-1269 enhances the proliferation and tumorigenicity of HCC cells. A. Real-time PCR analysis of miR-1269 in BEL-7404 and Huh7 cells stably overexpressing miR-1269. Transcript levels were normalized to U6 expression. B. The effects of miR-1269 overexpression on cell viability of the indicated cell lines analyzed by MTT assay. C. Representative micrographs (left panel) and quantification (right panel) of crystal violet stained colonies formed by the indicated cell lines. D. Representative micrographs (left panel) and quantification (right panel) of cell colonies determined by anchorage-independent growth ability assay. E. The effect of miR-1269 on cell cycle progression of the indicated cell lines analyzed by flow cytometry. Each bar represents the mean ± SD of three independent experiments. * P < 0.05.
Mentions: To explore the biological function of miR-1269 in HCC progression, BEL-7404 and Huh7 cells stably overexpressing miR-1269 were established (Figure 2A). Cell viability was then measured by MTT assay. A shown in Figure 2B, ectopic expression of miR-1269 increased the growth rate of both HCC cell lines. Colony formation assay showed that upregulation of miR-1269 promoted the colony formation capacity of BEL-7404 and Huh7 cells (Figure 2C). Consistently, anchorage-independent growth assay revealed that the cells stably expressing miR-1269 formed more and larger-sized colonies than the control cells (Figure 2D). Furthermore, cell cycle analysis by flow cytometry showed a dramatic decrease in the percentage of cells in the G1/G0 phase and an increase in the percentage of cells in the S phase in miR-1269-overexpressing cells (Figure 2E). Collectively, these results suggest that upregulation of miR-1269 enhanced the proliferation, tumorigenicity and cell cycle progression of HCC cells.Figure 2

Bottom Line: Furthermore, we demonstrated that FOXO1 was a direct target of miR-1269.Suppression of FOXO1 by miR-1269 was associated with dysregulation of p21, cyclin D1, phosphorylated Rb and Ki67 expression, thereby playing an essential role in the growth of HCC cells.Our study indicated that overexpression of miR-1269 promotes cell proliferation in HCC through directly suppressing FOXO1, and functions as an oncomiR in HCC.

View Article: PubMed Central - PubMed

Affiliation: Department of Hepatobiliary Surgery, the Second Affiliated Hospital of Guangzhou, Medical University, Guangzhou 510260, China. drxueping@medmail.com.cn.

ABSTRACT

Background: Hepatocellular carcinoma (HCC) is one of the most common malignancies and a major cause of cancer-related mortality in the world. MicroRNAs (miRNAs) are small, noncoding RNAs that play essential roles in various stages during cancer progression. The aim of the current study was to elucidate the role of miR-1269 in the pathogenesis of HCC.

Methods: The expression of miR-1269 in HCC cells and tissues were determined by Real-time PCR analysis. Cell viability, colony formation and anchorage-independent growth ability assays were performed to examine cell proliferative capacity and tumorigenicity. Flow cytometry analysis was conducted to determine cell cycle progression. The expression of p21, CyclinD1, phosphorylated Rb, Rb and FOXO1 were examined by Western blotting analysis. Luciferase assay was used to determine whether FOXO1 is the direct target of miR-1269.

Results: miR-1269 was upregulated in HCC cells and tissues. Ectopic miR-1269 expression promoted, but inhibition of miR-1269 reduced, proliferation, tumorigenicity and cell cycle progression of HCC cells. Furthermore, we demonstrated that FOXO1 was a direct target of miR-1269. Suppression of FOXO1 by miR-1269 was associated with dysregulation of p21, cyclin D1, phosphorylated Rb and Ki67 expression, thereby playing an essential role in the growth of HCC cells.

Conclusions: Our study indicated that overexpression of miR-1269 promotes cell proliferation in HCC through directly suppressing FOXO1, and functions as an oncomiR in HCC.

Show MeSH
Related in: MedlinePlus