Limits...
Sundew plant, a potential source of anti-inflammatory agents, selectively induces G2/M arrest and apoptosis in MCF-7 cells through upregulation of p53 and Bax/Bcl-2 ratio

View Article: PubMed Central - PubMed

ABSTRACT

The worldwide cancer incidences are remarkable despite the advancement in cancer drug discovery field, highlighting the need for new therapies focusing on cancer cell and its microenvironment, including inflammation. Several species of Drosera (family: Droseraceae) are used in various traditional as well as homeopathic systems of medicine. Drosera burmannii Vahl. is also enlisted in French Pharmacopoeia in 1965 for the treatment of inflammatory diseases, including chronic bronchitis, asthma and whooping cough. The present study is designed to substantiate the potential of D. burmannii in in vitro anticancer activity and its relation with anti-inflammatory property. In vitro anticancer study revealed that DBME is inhibiting the proliferation of MCF-7 cells without affecting the viability of other malignant and non-malignant cells. DBME induced G2/M phase arrest and apoptosis in MCF-7 cells by suppressing the expression of cyclin A1, cyclin B1 and Cdk-1 and increasing the expression of p53, Bax/Bcl-2 ratio leading to activation of caspases and PARP degradation. Presence of caspase-8 (Z-IETD-fmk) and caspase-9 (Z-LEHD-fmk) inhibitors alone did prevent the apoptosis partially while apoptosis prevention was significantly observed when used in combination, suggesting vital role of caspases in DBME-induced apoptosis in MCF-7 cells. DBME also downregulated LPS-induced increased expression of iNOS, COX-2 and TNF-α along with suppression on intracellular ROS production that confirms the potential of DBME as anti-inflammatory extract. GCMS analysis revealed the presence of four major compounds hexadecanoic acid, tetradecanoic acid, hexadecen-1-ol, trans-9 and 1-tetradecanol along with some other fatty acid derivatives and carotenoids (Beta-doradecin) in DBME. These findings confirmed the anti-inflammatory activity of DBME, which is already listed in French Pharmacopeia in 1965. Here we have additionally reported the anti-breast cancer activity of DBME and its relation to the anti-inflammatory potential. Hence, an ethnopharmacological approach can be considered as useful tool for the discovery of new drug leads.

No MeSH data available.


Related in: MedlinePlus

In vitro anti-inflammatory effects of DBME in RAW 264.7 cells. (a) Effect of DBME on cell proliferation and viability of murine macrophage RAW 264.7 cells. (b) Effect of DBME on LPS-induced production of nitrite. (c) Effect of DBME on LPS-induced production of TNF-α. (d) Effect of DBME on LPS-induced production of ROS. (e) Effect of DBME on LPS-induced expression of inflammatory proteins. (f) Effect of DBME on LPS-induced mRNA levels of inflammatory proteins in RAW 264.7 cells. Cytotoxicity data are expressed as mean±S.D. (N=6); *P<0.05, **P<0.01 and ***P<0.001 versus 0 μg/ml. Nitrite, TNF-α and ROS data are expressed as mean±S.D. (N=3); *P<0.05, **P<0.01 and ***P<0.001 versus 0 μg/ml LPS. #P<0.05, ##P<0.01 and ###P<0.001 versus 1 μg/ml LPS.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4979533&req=5

fig4: In vitro anti-inflammatory effects of DBME in RAW 264.7 cells. (a) Effect of DBME on cell proliferation and viability of murine macrophage RAW 264.7 cells. (b) Effect of DBME on LPS-induced production of nitrite. (c) Effect of DBME on LPS-induced production of TNF-α. (d) Effect of DBME on LPS-induced production of ROS. (e) Effect of DBME on LPS-induced expression of inflammatory proteins. (f) Effect of DBME on LPS-induced mRNA levels of inflammatory proteins in RAW 264.7 cells. Cytotoxicity data are expressed as mean±S.D. (N=6); *P<0.05, **P<0.01 and ***P<0.001 versus 0 μg/ml. Nitrite, TNF-α and ROS data are expressed as mean±S.D. (N=3); *P<0.05, **P<0.01 and ***P<0.001 versus 0 μg/ml LPS. #P<0.05, ##P<0.01 and ###P<0.001 versus 1 μg/ml LPS.

Mentions: Before determining the effect of DBME on LPS-induced NO, TNF-α and ROS production in RAW 264.7, the cytotoxicity of DBME on RAW 264.7 cells was examined using the WST-1 reagent. The cytotoxic effect was tested to establish the appropriate concentration ranges of DBME for the analysis of ongoing experiments (Figure 4a). The non-toxic concentrations (30, 50 and 80 μg/ml) were selected for further experiments. These concentrations significantly inhibited the LPS-induced nitrite (Figure 4b), TNF-α (Figure 4c) and ROS (Figure 4d) production in RAW 264.7 cells. After 24 h, LPS produced a considerable increase in NO and TNF-α in culture media and intracellular ROS. DBME drastically inhibited nitrite, TNF-α and ROS production in a dose-dependent manner, with a maximum effect at 80 μg/ml.


Sundew plant, a potential source of anti-inflammatory agents, selectively induces G2/M arrest and apoptosis in MCF-7 cells through upregulation of p53 and Bax/Bcl-2 ratio
In vitro anti-inflammatory effects of DBME in RAW 264.7 cells. (a) Effect of DBME on cell proliferation and viability of murine macrophage RAW 264.7 cells. (b) Effect of DBME on LPS-induced production of nitrite. (c) Effect of DBME on LPS-induced production of TNF-α. (d) Effect of DBME on LPS-induced production of ROS. (e) Effect of DBME on LPS-induced expression of inflammatory proteins. (f) Effect of DBME on LPS-induced mRNA levels of inflammatory proteins in RAW 264.7 cells. Cytotoxicity data are expressed as mean±S.D. (N=6); *P<0.05, **P<0.01 and ***P<0.001 versus 0 μg/ml. Nitrite, TNF-α and ROS data are expressed as mean±S.D. (N=3); *P<0.05, **P<0.01 and ***P<0.001 versus 0 μg/ml LPS. #P<0.05, ##P<0.01 and ###P<0.001 versus 1 μg/ml LPS.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4979533&req=5

fig4: In vitro anti-inflammatory effects of DBME in RAW 264.7 cells. (a) Effect of DBME on cell proliferation and viability of murine macrophage RAW 264.7 cells. (b) Effect of DBME on LPS-induced production of nitrite. (c) Effect of DBME on LPS-induced production of TNF-α. (d) Effect of DBME on LPS-induced production of ROS. (e) Effect of DBME on LPS-induced expression of inflammatory proteins. (f) Effect of DBME on LPS-induced mRNA levels of inflammatory proteins in RAW 264.7 cells. Cytotoxicity data are expressed as mean±S.D. (N=6); *P<0.05, **P<0.01 and ***P<0.001 versus 0 μg/ml. Nitrite, TNF-α and ROS data are expressed as mean±S.D. (N=3); *P<0.05, **P<0.01 and ***P<0.001 versus 0 μg/ml LPS. #P<0.05, ##P<0.01 and ###P<0.001 versus 1 μg/ml LPS.
Mentions: Before determining the effect of DBME on LPS-induced NO, TNF-α and ROS production in RAW 264.7, the cytotoxicity of DBME on RAW 264.7 cells was examined using the WST-1 reagent. The cytotoxic effect was tested to establish the appropriate concentration ranges of DBME for the analysis of ongoing experiments (Figure 4a). The non-toxic concentrations (30, 50 and 80 μg/ml) were selected for further experiments. These concentrations significantly inhibited the LPS-induced nitrite (Figure 4b), TNF-α (Figure 4c) and ROS (Figure 4d) production in RAW 264.7 cells. After 24 h, LPS produced a considerable increase in NO and TNF-α in culture media and intracellular ROS. DBME drastically inhibited nitrite, TNF-α and ROS production in a dose-dependent manner, with a maximum effect at 80 μg/ml.

View Article: PubMed Central - PubMed

ABSTRACT

The worldwide cancer incidences are remarkable despite the advancement in cancer drug discovery field, highlighting the need for new therapies focusing on cancer cell and its microenvironment, including inflammation. Several species of Drosera (family: Droseraceae) are used in various traditional as well as homeopathic systems of medicine. Drosera burmannii Vahl. is also enlisted in French Pharmacopoeia in 1965 for the treatment of inflammatory diseases, including chronic bronchitis, asthma and whooping cough. The present study is designed to substantiate the potential of D. burmannii in in vitro anticancer activity and its relation with anti-inflammatory property. In vitro anticancer study revealed that DBME is inhibiting the proliferation of MCF-7 cells without affecting the viability of other malignant and non-malignant cells. DBME induced G2/M phase arrest and apoptosis in MCF-7 cells by suppressing the expression of cyclin A1, cyclin B1 and Cdk-1 and increasing the expression of p53, Bax/Bcl-2 ratio leading to activation of caspases and PARP degradation. Presence of caspase-8 (Z-IETD-fmk) and caspase-9 (Z-LEHD-fmk) inhibitors alone did prevent the apoptosis partially while apoptosis prevention was significantly observed when used in combination, suggesting vital role of caspases in DBME-induced apoptosis in MCF-7 cells. DBME also downregulated LPS-induced increased expression of iNOS, COX-2 and TNF-&alpha; along with suppression on intracellular ROS production that confirms the potential of DBME as anti-inflammatory extract. GCMS analysis revealed the presence of four major compounds hexadecanoic acid, tetradecanoic acid, hexadecen-1-ol, trans-9 and 1-tetradecanol along with some other fatty acid derivatives and carotenoids (Beta-doradecin) in DBME. These findings confirmed the anti-inflammatory activity of DBME, which is already listed in French Pharmacopeia in 1965. Here we have additionally reported the anti-breast cancer activity of DBME and its relation to the anti-inflammatory potential. Hence, an ethnopharmacological approach can be considered as useful tool for the discovery of new drug leads.

No MeSH data available.


Related in: MedlinePlus