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Hedyotis diffusa Willd. extract suppresses proliferation and induces apoptosis via IL-6-inducible STAT3 pathway inactivation in human colorectal cancer cells.

Lin J, Li Q, Chen H, Lin H, Lai Z, Peng J - Oncol Lett (2015)

Bottom Line: Pretreatment of HT-29 cells with IL-6 led to an increase in cell viability, colony formation and phosphorylated STAT3 (p-STAT3) expression.Treatment of these cells with EEHDW prior to IL-6 stimulation resulted in a significant reduction in the IL-6-induced phosphorylation of STAT3.In addition, EEHDW treatment significantly reduced the mRNA expression levels of cyclin D1, cyclin-dependent kinase 4 and B-cell lymphoma-2 (Bcl-2), and upregulated the expression levels of Bcl-2-associated X protein (P<0.05), which are important target genes of the IL-6/STAT3 pathway.

View Article: PubMed Central - PubMed

Affiliation: Academy of Integrative Medicine Biomedical Research Center, Fujian University of Traditional Chinese Medicine, Fuzhou, Fujian 350122, P.R. China ; Fujian Key Laboratory of Integrative Medicine on Geriatrics, Fujian University of Traditional Chinese Medicine, Fuzhou, Fujian 350122, P.R. China.

ABSTRACT

Recent studies have indicated that the inflammatory microenvironment plays a significant role in colorectal cancer (CRC). The interleukin-6/signal transducer and activator of transcription 3 (IL-6/STAT3) signaling pathway mediates the proliferative and anti-apoptotic activities required for oncogenesis under inflammatory conditions; thus, suppressing tumor growth by targeting the IL-6/STAT3 pathway is a promising therapeutic strategy for CRC. Our previous study reported that the ethanol extract obtained from Hedyotis diffusa Willd. (EEHDW) can induce apoptosis, and inhibit the proliferation of colon cancer cells and tumor angiogenesis by modulating various signaling pathways; however, less is known regarding the activity of EEHDW in a cancer-promoting inflammatory environment. Therefore, the present study investigated whether EEHDW inhibits the growth of the CRC HT-29 cell line via the IL-6/STAT3 signaling pathway. Pretreatment of HT-29 cells with IL-6 led to an increase in cell viability, colony formation and phosphorylated STAT3 (p-STAT3) expression. Treatment of these cells with EEHDW prior to IL-6 stimulation resulted in a significant reduction in the IL-6-induced phosphorylation of STAT3. In addition, EEHDW treatment significantly reduced the mRNA expression levels of cyclin D1, cyclin-dependent kinase 4 and B-cell lymphoma-2 (Bcl-2), and upregulated the expression levels of Bcl-2-associated X protein (P<0.05), which are important target genes of the IL-6/STAT3 pathway. These findings strongly indicated that EEHDW suppresses tumor cell growth and induces the apoptosis of human CRC cells via inactivation of the IL-6/STAT3 signaling pathway.

No MeSH data available.


Related in: MedlinePlus

Cellular morphology of EEHDW-treated HT-29 cells. The HT-29 cells were pretreated with the indicated doses of EEHDW for 1 h prior to IL-6 stimulation for 24 h. Changes in cellular morphology were visualized by DAPI staining, and images of the control and treated cells were captured using a phase-contrast fluorescence microscope (magnification, ×200). Images are representative of three independent experiments. IL-6, interleukin-6; EEHDW, ethanol extract obtained from Hedyotis diffusa Willd.
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f4-ol-09-04-1962: Cellular morphology of EEHDW-treated HT-29 cells. The HT-29 cells were pretreated with the indicated doses of EEHDW for 1 h prior to IL-6 stimulation for 24 h. Changes in cellular morphology were visualized by DAPI staining, and images of the control and treated cells were captured using a phase-contrast fluorescence microscope (magnification, ×200). Images are representative of three independent experiments. IL-6, interleukin-6; EEHDW, ethanol extract obtained from Hedyotis diffusa Willd.

Mentions: Our previous study observed that EEHDW induced apoptosis in HT-29 cells in the absence of IL-6 stimulation (20). To determine whether EEHDEW induces cell apoptosis during cytokine-mediated activation and proliferation, the induction of apoptosis in the IL-6-stimulated HT-29 cells was determined by performing Annexin V/PI staining and FACS analysis (Fig. 3A and B). In comparison to the unstimulated cells, stimulation with 10 ng/ml IL-6 did not significantly alter the proportion of the apoptotic cells (P>0.05). By contrast, EEHDW treatment significantly increased the percentage of cells undergoing early and late apoptosis in a dose-dependent manner (P<0.05 vs. cells stimulated with IL-6 alone). In addition, the cellular morphology and extent of DNA condensation of the apoptotic HT-29 cells were examined by DAPI staining (Fig. 4). Nuclei staining of the HT-29 cells treated with EEHDW was more intense than the untreated cells, indicating that EEHDW promotes HT-29 cell apoptosis in the presence of IL-6 stimulation.


Hedyotis diffusa Willd. extract suppresses proliferation and induces apoptosis via IL-6-inducible STAT3 pathway inactivation in human colorectal cancer cells.

Lin J, Li Q, Chen H, Lin H, Lai Z, Peng J - Oncol Lett (2015)

Cellular morphology of EEHDW-treated HT-29 cells. The HT-29 cells were pretreated with the indicated doses of EEHDW for 1 h prior to IL-6 stimulation for 24 h. Changes in cellular morphology were visualized by DAPI staining, and images of the control and treated cells were captured using a phase-contrast fluorescence microscope (magnification, ×200). Images are representative of three independent experiments. IL-6, interleukin-6; EEHDW, ethanol extract obtained from Hedyotis diffusa Willd.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4356405&req=5

f4-ol-09-04-1962: Cellular morphology of EEHDW-treated HT-29 cells. The HT-29 cells were pretreated with the indicated doses of EEHDW for 1 h prior to IL-6 stimulation for 24 h. Changes in cellular morphology were visualized by DAPI staining, and images of the control and treated cells were captured using a phase-contrast fluorescence microscope (magnification, ×200). Images are representative of three independent experiments. IL-6, interleukin-6; EEHDW, ethanol extract obtained from Hedyotis diffusa Willd.
Mentions: Our previous study observed that EEHDW induced apoptosis in HT-29 cells in the absence of IL-6 stimulation (20). To determine whether EEHDEW induces cell apoptosis during cytokine-mediated activation and proliferation, the induction of apoptosis in the IL-6-stimulated HT-29 cells was determined by performing Annexin V/PI staining and FACS analysis (Fig. 3A and B). In comparison to the unstimulated cells, stimulation with 10 ng/ml IL-6 did not significantly alter the proportion of the apoptotic cells (P>0.05). By contrast, EEHDW treatment significantly increased the percentage of cells undergoing early and late apoptosis in a dose-dependent manner (P<0.05 vs. cells stimulated with IL-6 alone). In addition, the cellular morphology and extent of DNA condensation of the apoptotic HT-29 cells were examined by DAPI staining (Fig. 4). Nuclei staining of the HT-29 cells treated with EEHDW was more intense than the untreated cells, indicating that EEHDW promotes HT-29 cell apoptosis in the presence of IL-6 stimulation.

Bottom Line: Pretreatment of HT-29 cells with IL-6 led to an increase in cell viability, colony formation and phosphorylated STAT3 (p-STAT3) expression.Treatment of these cells with EEHDW prior to IL-6 stimulation resulted in a significant reduction in the IL-6-induced phosphorylation of STAT3.In addition, EEHDW treatment significantly reduced the mRNA expression levels of cyclin D1, cyclin-dependent kinase 4 and B-cell lymphoma-2 (Bcl-2), and upregulated the expression levels of Bcl-2-associated X protein (P<0.05), which are important target genes of the IL-6/STAT3 pathway.

View Article: PubMed Central - PubMed

Affiliation: Academy of Integrative Medicine Biomedical Research Center, Fujian University of Traditional Chinese Medicine, Fuzhou, Fujian 350122, P.R. China ; Fujian Key Laboratory of Integrative Medicine on Geriatrics, Fujian University of Traditional Chinese Medicine, Fuzhou, Fujian 350122, P.R. China.

ABSTRACT

Recent studies have indicated that the inflammatory microenvironment plays a significant role in colorectal cancer (CRC). The interleukin-6/signal transducer and activator of transcription 3 (IL-6/STAT3) signaling pathway mediates the proliferative and anti-apoptotic activities required for oncogenesis under inflammatory conditions; thus, suppressing tumor growth by targeting the IL-6/STAT3 pathway is a promising therapeutic strategy for CRC. Our previous study reported that the ethanol extract obtained from Hedyotis diffusa Willd. (EEHDW) can induce apoptosis, and inhibit the proliferation of colon cancer cells and tumor angiogenesis by modulating various signaling pathways; however, less is known regarding the activity of EEHDW in a cancer-promoting inflammatory environment. Therefore, the present study investigated whether EEHDW inhibits the growth of the CRC HT-29 cell line via the IL-6/STAT3 signaling pathway. Pretreatment of HT-29 cells with IL-6 led to an increase in cell viability, colony formation and phosphorylated STAT3 (p-STAT3) expression. Treatment of these cells with EEHDW prior to IL-6 stimulation resulted in a significant reduction in the IL-6-induced phosphorylation of STAT3. In addition, EEHDW treatment significantly reduced the mRNA expression levels of cyclin D1, cyclin-dependent kinase 4 and B-cell lymphoma-2 (Bcl-2), and upregulated the expression levels of Bcl-2-associated X protein (P<0.05), which are important target genes of the IL-6/STAT3 pathway. These findings strongly indicated that EEHDW suppresses tumor cell growth and induces the apoptosis of human CRC cells via inactivation of the IL-6/STAT3 signaling pathway.

No MeSH data available.


Related in: MedlinePlus