Limits...
miR-145 mediates the antiproliferative and gene regulatory effects of vitamin D3 by directly targeting E2F3 in gastric cancer cells.

Chang S, Gao L, Yang Y, Tong D, Guo B, Liu L, Li Z, Song T, Huang C - Oncotarget (2015)

Bottom Line: Inhibition of miR-145 reverses the antiproliferative effect of 1,25(OH)2D3.Furthermore, miR-145 expression was lower in tumors compared with matched normal samples and correlated with increased the E2F3 transcription factor protein staining.Altogether, our results indicate that miR-145 mediates the antiproliferative and gene regulatory effects of vitamin D3 in GC cells and might hold promise for prognosis and therapeutic strategies for GC treatment.

View Article: PubMed Central - PubMed

Affiliation: Department of Genetics and Molecular Biology/Key Laboratory of Environment and Genes Related to Diseases, College of Medicine, Xi'an Jiaotong University, Xi'an, Shaanxi, P. R. China.

ABSTRACT
VitaminD3 signaling is involved in inhibiting the development and progression of gastric cancer (GC), while the active vitamin D metabolite 1-alpha,25-dihydroxyvitamin D3 (1,25(OH)2D3)-mediated gene regulatory mechanisms in GC remain unclear. We found that miR-145 is induced by 1,25(OH)2D3 in a dose- and vitamin D receptor (VDR)-dependent manner in GC cells. Inhibition of miR-145 reverses the antiproliferative effect of 1,25(OH)2D3. Furthermore, miR-145 expression was lower in tumors compared with matched normal samples and correlated with increased the E2F3 transcription factor protein staining. Overexpression of miR-145 inhibited colony formation, cell viability and induced cell arrest in S-phase in GC cells by targeting E2F3 and CDK6. miR-145 inhibition consistently abrogates the 1,25(OH)2D3-mediated suppression of E2F3, CDK6, CDK2 and CCNA2 genes. Altogether, our results indicate that miR-145 mediates the antiproliferative and gene regulatory effects of vitamin D3 in GC cells and might hold promise for prognosis and therapeutic strategies for GC treatment.

No MeSH data available.


Related in: MedlinePlus

1,25(OH)2D3 inhibits GC cell proliferation and promote cell apoptosis in vitro(A–B) SGC-7901 and AGS cells were treated with various doses of 1,25(OH)2D3 and the effects were determined by MTT assay after 24, 48 and 72 hours. (C–D) SGC-7901 and AGS cells were treated with 500 nM 1,25(OH)2D3 for 48 hours and cell cycle distribution was analyzed by flow cytometry. Histogram indicated the percentage of cells in G1, S and G2 cell-cycle phases. (E–F) SGC-7901 and AGS cells were treated with 500 nM 1,25(OH)2D3 for 48 hours, apoptosis was determined by Annexin V staining and flow cytometry. The experiments have been repeated 3 times, representative results of 3 independent experiments were shown. Data shown are mean values. (*P < 0.05; **P < 0.01; ***p < 0.001.)
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4480708&req=5

Figure 1: 1,25(OH)2D3 inhibits GC cell proliferation and promote cell apoptosis in vitro(A–B) SGC-7901 and AGS cells were treated with various doses of 1,25(OH)2D3 and the effects were determined by MTT assay after 24, 48 and 72 hours. (C–D) SGC-7901 and AGS cells were treated with 500 nM 1,25(OH)2D3 for 48 hours and cell cycle distribution was analyzed by flow cytometry. Histogram indicated the percentage of cells in G1, S and G2 cell-cycle phases. (E–F) SGC-7901 and AGS cells were treated with 500 nM 1,25(OH)2D3 for 48 hours, apoptosis was determined by Annexin V staining and flow cytometry. The experiments have been repeated 3 times, representative results of 3 independent experiments were shown. Data shown are mean values. (*P < 0.05; **P < 0.01; ***p < 0.001.)

Mentions: To examine the effects of 1,25(OH)2D3 on GC cells, SGC-7901 and AGS cells were treated with various doses of 1,25(OH)2D3. The MTT assay indicated that 1,25(OH)2D3 significantly suppressed cell growth and mostly reached a plateau at the 500 nmol dose point when compared with the control group in vitro in both cell lines (Figure 1A and 1B). We then studied the potential mechanism of 1,25(OH)2D3-induced growth suppression. The cells were incubated in serum-free medium to synchronize them in the G1 phase. 1,25(OH)2D3 slightly decreased the percentage of cells in the S phase in SGC-7901 cells while no obvious change in AGS cells (Figure 1C and 1D). In addition, annexin V staining analysis dispalyed that 1,25(OH)2D3 promoted cancer cell apoptosis, which is consistent with the study of vitamin D-induced apoptosis through PTEN upregulation [19] (Figure 1E and 1F). Based on these findings, we concluded that 1,25(OH)2D3 could regulate GC cell proliferation and apoptosis.


miR-145 mediates the antiproliferative and gene regulatory effects of vitamin D3 by directly targeting E2F3 in gastric cancer cells.

Chang S, Gao L, Yang Y, Tong D, Guo B, Liu L, Li Z, Song T, Huang C - Oncotarget (2015)

1,25(OH)2D3 inhibits GC cell proliferation and promote cell apoptosis in vitro(A–B) SGC-7901 and AGS cells were treated with various doses of 1,25(OH)2D3 and the effects were determined by MTT assay after 24, 48 and 72 hours. (C–D) SGC-7901 and AGS cells were treated with 500 nM 1,25(OH)2D3 for 48 hours and cell cycle distribution was analyzed by flow cytometry. Histogram indicated the percentage of cells in G1, S and G2 cell-cycle phases. (E–F) SGC-7901 and AGS cells were treated with 500 nM 1,25(OH)2D3 for 48 hours, apoptosis was determined by Annexin V staining and flow cytometry. The experiments have been repeated 3 times, representative results of 3 independent experiments were shown. Data shown are mean values. (*P < 0.05; **P < 0.01; ***p < 0.001.)
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4480708&req=5

Figure 1: 1,25(OH)2D3 inhibits GC cell proliferation and promote cell apoptosis in vitro(A–B) SGC-7901 and AGS cells were treated with various doses of 1,25(OH)2D3 and the effects were determined by MTT assay after 24, 48 and 72 hours. (C–D) SGC-7901 and AGS cells were treated with 500 nM 1,25(OH)2D3 for 48 hours and cell cycle distribution was analyzed by flow cytometry. Histogram indicated the percentage of cells in G1, S and G2 cell-cycle phases. (E–F) SGC-7901 and AGS cells were treated with 500 nM 1,25(OH)2D3 for 48 hours, apoptosis was determined by Annexin V staining and flow cytometry. The experiments have been repeated 3 times, representative results of 3 independent experiments were shown. Data shown are mean values. (*P < 0.05; **P < 0.01; ***p < 0.001.)
Mentions: To examine the effects of 1,25(OH)2D3 on GC cells, SGC-7901 and AGS cells were treated with various doses of 1,25(OH)2D3. The MTT assay indicated that 1,25(OH)2D3 significantly suppressed cell growth and mostly reached a plateau at the 500 nmol dose point when compared with the control group in vitro in both cell lines (Figure 1A and 1B). We then studied the potential mechanism of 1,25(OH)2D3-induced growth suppression. The cells were incubated in serum-free medium to synchronize them in the G1 phase. 1,25(OH)2D3 slightly decreased the percentage of cells in the S phase in SGC-7901 cells while no obvious change in AGS cells (Figure 1C and 1D). In addition, annexin V staining analysis dispalyed that 1,25(OH)2D3 promoted cancer cell apoptosis, which is consistent with the study of vitamin D-induced apoptosis through PTEN upregulation [19] (Figure 1E and 1F). Based on these findings, we concluded that 1,25(OH)2D3 could regulate GC cell proliferation and apoptosis.

Bottom Line: Inhibition of miR-145 reverses the antiproliferative effect of 1,25(OH)2D3.Furthermore, miR-145 expression was lower in tumors compared with matched normal samples and correlated with increased the E2F3 transcription factor protein staining.Altogether, our results indicate that miR-145 mediates the antiproliferative and gene regulatory effects of vitamin D3 in GC cells and might hold promise for prognosis and therapeutic strategies for GC treatment.

View Article: PubMed Central - PubMed

Affiliation: Department of Genetics and Molecular Biology/Key Laboratory of Environment and Genes Related to Diseases, College of Medicine, Xi'an Jiaotong University, Xi'an, Shaanxi, P. R. China.

ABSTRACT
VitaminD3 signaling is involved in inhibiting the development and progression of gastric cancer (GC), while the active vitamin D metabolite 1-alpha,25-dihydroxyvitamin D3 (1,25(OH)2D3)-mediated gene regulatory mechanisms in GC remain unclear. We found that miR-145 is induced by 1,25(OH)2D3 in a dose- and vitamin D receptor (VDR)-dependent manner in GC cells. Inhibition of miR-145 reverses the antiproliferative effect of 1,25(OH)2D3. Furthermore, miR-145 expression was lower in tumors compared with matched normal samples and correlated with increased the E2F3 transcription factor protein staining. Overexpression of miR-145 inhibited colony formation, cell viability and induced cell arrest in S-phase in GC cells by targeting E2F3 and CDK6. miR-145 inhibition consistently abrogates the 1,25(OH)2D3-mediated suppression of E2F3, CDK6, CDK2 and CCNA2 genes. Altogether, our results indicate that miR-145 mediates the antiproliferative and gene regulatory effects of vitamin D3 in GC cells and might hold promise for prognosis and therapeutic strategies for GC treatment.

No MeSH data available.


Related in: MedlinePlus