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Juglone exerts antitumor effect in ovarian cancer cells.

Fang F, Qin Y, Qi L, Fang Q, Zhao L, Chen S, Li Q, Zhang D, Wang L - Iran J Basic Med Sci (2015)

Bottom Line: Juglone significantly inhibited SKOV3 cell proliferation as shown by G0/G1 phase arrest, and this effect was mediated by inactivation of cyclin D1 protein (P<0.05).Juglone sufficiently inhibited invasion while evidently decreased MMP-2 expression (P<0.05).The results suggest that juglone could probably induce apoptosis through mitochondrial pathway and restrained cell invasiveness by decreasing MMP expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, Jilin Medical College, Jilin, Jilin, China.

ABSTRACT

Objectives: Juglone is isolated from many species of the Juglandaceae family and used as an anti-viral, anti-bacterial, and anti-tumor therapeutic. Here, we evaluated juglone-induced antitumor effect in ovarian cancer SKOV3 cells.

Materials and methods: MTT assay was performed to examine juglone anti-proliferative effect. Cell cycle and apoptosis were studied using flow cytometry in juglone-treated SKOV3 cells. To investigate molecular mechanism of cell cycle and apoptosis, protein expression levels were measured by Western blot analysis of cyclin D1, Bcl-2, Bax, cytochrome c, caspase-9 and caspase-3. To investigate the motility of juglone-treated SKOV3 cell, Matrigel invasion assay was employed to characterize cell invasion. Also, matrix metalloproteinase-2 (MMP-2) expression levels were detected by western blot.

Results: Juglone significantly inhibited SKOV3 cell proliferation as shown by G0/G1 phase arrest, and this effect was mediated by inactivation of cyclin D1 protein (P<0.05). Juglone induced apoptosis in SKOV3 cell which was accompanied by caspase-9 and caspase-3 activation (P<0.05). Juglone decreased Bcl-2 levels and increased Bax and cytochrome c (Cyt c) levels (P<0.05). Juglone sufficiently inhibited invasion while evidently decreased MMP-2 expression (P<0.05).

Conclusion: The results suggest that juglone could probably induce apoptosis through mitochondrial pathway and restrained cell invasiveness by decreasing MMP expression.

No MeSH data available.


Related in: MedlinePlus

Analysis of juglone-treated SKOV3 cells apoptosis. A: Cells were treated with juglone for 24 hr, then stained with Annexin V-FITC and PI in the dark, and analyzed using flow cytometry. B and C: Western blot analysis was performed with antibodies against Cyt c, Bcl-2, Bax, procaspase-9, and activated caspase-3 in juglone-treated SKOV3 cells. β-actin was used as an internal reference. *P <0.05, **P<0.01 compared to control
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Figure 3: Analysis of juglone-treated SKOV3 cells apoptosis. A: Cells were treated with juglone for 24 hr, then stained with Annexin V-FITC and PI in the dark, and analyzed using flow cytometry. B and C: Western blot analysis was performed with antibodies against Cyt c, Bcl-2, Bax, procaspase-9, and activated caspase-3 in juglone-treated SKOV3 cells. β-actin was used as an internal reference. *P <0.05, **P<0.01 compared to control

Mentions: To quantitate apoptosis, we performed Annexin V-FITC/PI double staining. The results showed significant early and late stage apoptotic cell accumulation in juglone-treated cells in dose-dependent manners (Figure 3A). Apoptosis is always accompanied by mitochondrial membrane disruption that leads to Cyt c release from the mitochondria into the cytoplasm (7). As shown in Figure 3B, we found that Cyt c protein increased following incubation with juglone. Bcl-2 and Bax belong to a multi-gene protein family that plays an important role in regulating apoptosis. Bcl-2 is an anti-apoptotic protein, whereas Bax is a pro-apoptotic protein and antagonizes Bcl-2 effects (7). As shown in Figure 3B, anti-apoptotic Bcl-2 protein levels declined and pro-apoptotic Bax protein levels increased in response to juglone in a dose-dependent manner. Thus, the Bax/Bcl-2 ratio increased. Caspase, a family of cysteine proteases, forms integral parts of the apoptosis pathway. Therefore, we detected caspase-9 and caspase-3 protein levels.


Juglone exerts antitumor effect in ovarian cancer cells.

Fang F, Qin Y, Qi L, Fang Q, Zhao L, Chen S, Li Q, Zhang D, Wang L - Iran J Basic Med Sci (2015)

Analysis of juglone-treated SKOV3 cells apoptosis. A: Cells were treated with juglone for 24 hr, then stained with Annexin V-FITC and PI in the dark, and analyzed using flow cytometry. B and C: Western blot analysis was performed with antibodies against Cyt c, Bcl-2, Bax, procaspase-9, and activated caspase-3 in juglone-treated SKOV3 cells. β-actin was used as an internal reference. *P <0.05, **P<0.01 compared to control
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4509948&req=5

Figure 3: Analysis of juglone-treated SKOV3 cells apoptosis. A: Cells were treated with juglone for 24 hr, then stained with Annexin V-FITC and PI in the dark, and analyzed using flow cytometry. B and C: Western blot analysis was performed with antibodies against Cyt c, Bcl-2, Bax, procaspase-9, and activated caspase-3 in juglone-treated SKOV3 cells. β-actin was used as an internal reference. *P <0.05, **P<0.01 compared to control
Mentions: To quantitate apoptosis, we performed Annexin V-FITC/PI double staining. The results showed significant early and late stage apoptotic cell accumulation in juglone-treated cells in dose-dependent manners (Figure 3A). Apoptosis is always accompanied by mitochondrial membrane disruption that leads to Cyt c release from the mitochondria into the cytoplasm (7). As shown in Figure 3B, we found that Cyt c protein increased following incubation with juglone. Bcl-2 and Bax belong to a multi-gene protein family that plays an important role in regulating apoptosis. Bcl-2 is an anti-apoptotic protein, whereas Bax is a pro-apoptotic protein and antagonizes Bcl-2 effects (7). As shown in Figure 3B, anti-apoptotic Bcl-2 protein levels declined and pro-apoptotic Bax protein levels increased in response to juglone in a dose-dependent manner. Thus, the Bax/Bcl-2 ratio increased. Caspase, a family of cysteine proteases, forms integral parts of the apoptosis pathway. Therefore, we detected caspase-9 and caspase-3 protein levels.

Bottom Line: Juglone significantly inhibited SKOV3 cell proliferation as shown by G0/G1 phase arrest, and this effect was mediated by inactivation of cyclin D1 protein (P<0.05).Juglone sufficiently inhibited invasion while evidently decreased MMP-2 expression (P<0.05).The results suggest that juglone could probably induce apoptosis through mitochondrial pathway and restrained cell invasiveness by decreasing MMP expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, Jilin Medical College, Jilin, Jilin, China.

ABSTRACT

Objectives: Juglone is isolated from many species of the Juglandaceae family and used as an anti-viral, anti-bacterial, and anti-tumor therapeutic. Here, we evaluated juglone-induced antitumor effect in ovarian cancer SKOV3 cells.

Materials and methods: MTT assay was performed to examine juglone anti-proliferative effect. Cell cycle and apoptosis were studied using flow cytometry in juglone-treated SKOV3 cells. To investigate molecular mechanism of cell cycle and apoptosis, protein expression levels were measured by Western blot analysis of cyclin D1, Bcl-2, Bax, cytochrome c, caspase-9 and caspase-3. To investigate the motility of juglone-treated SKOV3 cell, Matrigel invasion assay was employed to characterize cell invasion. Also, matrix metalloproteinase-2 (MMP-2) expression levels were detected by western blot.

Results: Juglone significantly inhibited SKOV3 cell proliferation as shown by G0/G1 phase arrest, and this effect was mediated by inactivation of cyclin D1 protein (P<0.05). Juglone induced apoptosis in SKOV3 cell which was accompanied by caspase-9 and caspase-3 activation (P<0.05). Juglone decreased Bcl-2 levels and increased Bax and cytochrome c (Cyt c) levels (P<0.05). Juglone sufficiently inhibited invasion while evidently decreased MMP-2 expression (P<0.05).

Conclusion: The results suggest that juglone could probably induce apoptosis through mitochondrial pathway and restrained cell invasiveness by decreasing MMP expression.

No MeSH data available.


Related in: MedlinePlus