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In vitro and in vivo antitumor activity of a novel semisynthetic derivative of cucurbitacin B.

Silva IT, Carvalho A, Lang KL, Dudek SE, Masemann D, Durán FJ, Caro MS, Rapp UR, Wixler V, Schenkel EP, Simões CM, Ludwig S - PLoS ONE (2015)

Bottom Line: Despite the outstanding progress made in anti-tumor therapy, discovering effective anti-tumor drugs is still a challenging task.DACE arrested the cell cycle of lung epithelial cells at the G2/M phase and induced cell apoptosis by interfering with EGFR activation and its downstream signaling, including AKT, ERK, and STAT3.Taken together, these findings suggest that DACE is a promising lead compound for the development of an anti-lung-cancer drug.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmaceutical Sciences, Universidade Federal de Santa Catarina, Florianópolis, SC, Brazil; Institute of Molecular Virology (IMV), Center of Molecular Biology of Inflammation (ZMBE), Westfaelische-Wilhelms-University, Muenster, Germany.

ABSTRACT
Lung cancer is the most deadly type of cancer in humans, with non-small-cell lung cancer (NSCLC) being the most frequent and aggressive type of lung cancer showing high resistance to radiation and chemotherapy. Despite the outstanding progress made in anti-tumor therapy, discovering effective anti-tumor drugs is still a challenging task. Here we describe a new semisynthetic derivative of cucurbitacin B (DACE) as a potent inhibitor of NSCLC cell proliferation. DACE arrested the cell cycle of lung epithelial cells at the G2/M phase and induced cell apoptosis by interfering with EGFR activation and its downstream signaling, including AKT, ERK, and STAT3. Consistent with our in vitro studies, intraperitoneal application of DACE significantly suppressed the growth of mouse NSCLC that arises from type II alveolar pneumocytes due to constitutive expression of a human oncogenic c-RAF kinase (c-RAF-1-BxB) transgene in these cells. Taken together, these findings suggest that DACE is a promising lead compound for the development of an anti-lung-cancer drug.

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Related in: MedlinePlus

Cell and clonogenic growth inhibition by DACE.(A) Human non-small cell lung cancer cells (A549 cells) were treated with different concentrations of the DACE for 48h and 72h. The growth inhibition effects were determined by MTT assay and the IC50 was calculated by GraphPad Prism 5 software through a nonlinear fit-curve (log of compound concentration versus normalized response—variable slope). (B) A549 cells were treated with 0.5 and 1 μM of the DACE for 48h, followed by two washes for compound removal. Then, cells were plated for clonogenic assay; left—representative images of colonies formed from A549 cells under the different treatment conditions; right—number of colonies after 12 days. ***p<0.001 as compared with control.
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pone.0117794.g002: Cell and clonogenic growth inhibition by DACE.(A) Human non-small cell lung cancer cells (A549 cells) were treated with different concentrations of the DACE for 48h and 72h. The growth inhibition effects were determined by MTT assay and the IC50 was calculated by GraphPad Prism 5 software through a nonlinear fit-curve (log of compound concentration versus normalized response—variable slope). (B) A549 cells were treated with 0.5 and 1 μM of the DACE for 48h, followed by two washes for compound removal. Then, cells were plated for clonogenic assay; left—representative images of colonies formed from A549 cells under the different treatment conditions; right—number of colonies after 12 days. ***p<0.001 as compared with control.

Mentions: Initially, we explored whether DACE affects the proliferation of A549 human lung adenocarcinoma cells using MTT assay. Results of Fig. 2A show that DACE potently inhibited cellular proliferation in a concentration- and time-dependent manner. The calculated IC50 values for 48h and 72h were 0.42μM and 0.12μM, respectively (equivalent to log value of 2.62 and 2.075). To test the long term effect of DACE treatment, A549 cells were treated for 48h and then replated with low density into new plates without further treatment. Quantification of colonies formed twelve days after DACE treatment showed that their number was reduced compared to control cells to 64% and 76%, respectively, in both tested concentrations (p<0.001 vs. control) (Fig. 2B).


In vitro and in vivo antitumor activity of a novel semisynthetic derivative of cucurbitacin B.

Silva IT, Carvalho A, Lang KL, Dudek SE, Masemann D, Durán FJ, Caro MS, Rapp UR, Wixler V, Schenkel EP, Simões CM, Ludwig S - PLoS ONE (2015)

Cell and clonogenic growth inhibition by DACE.(A) Human non-small cell lung cancer cells (A549 cells) were treated with different concentrations of the DACE for 48h and 72h. The growth inhibition effects were determined by MTT assay and the IC50 was calculated by GraphPad Prism 5 software through a nonlinear fit-curve (log of compound concentration versus normalized response—variable slope). (B) A549 cells were treated with 0.5 and 1 μM of the DACE for 48h, followed by two washes for compound removal. Then, cells were plated for clonogenic assay; left—representative images of colonies formed from A549 cells under the different treatment conditions; right—number of colonies after 12 days. ***p<0.001 as compared with control.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4326133&req=5

pone.0117794.g002: Cell and clonogenic growth inhibition by DACE.(A) Human non-small cell lung cancer cells (A549 cells) were treated with different concentrations of the DACE for 48h and 72h. The growth inhibition effects were determined by MTT assay and the IC50 was calculated by GraphPad Prism 5 software through a nonlinear fit-curve (log of compound concentration versus normalized response—variable slope). (B) A549 cells were treated with 0.5 and 1 μM of the DACE for 48h, followed by two washes for compound removal. Then, cells were plated for clonogenic assay; left—representative images of colonies formed from A549 cells under the different treatment conditions; right—number of colonies after 12 days. ***p<0.001 as compared with control.
Mentions: Initially, we explored whether DACE affects the proliferation of A549 human lung adenocarcinoma cells using MTT assay. Results of Fig. 2A show that DACE potently inhibited cellular proliferation in a concentration- and time-dependent manner. The calculated IC50 values for 48h and 72h were 0.42μM and 0.12μM, respectively (equivalent to log value of 2.62 and 2.075). To test the long term effect of DACE treatment, A549 cells were treated for 48h and then replated with low density into new plates without further treatment. Quantification of colonies formed twelve days after DACE treatment showed that their number was reduced compared to control cells to 64% and 76%, respectively, in both tested concentrations (p<0.001 vs. control) (Fig. 2B).

Bottom Line: Despite the outstanding progress made in anti-tumor therapy, discovering effective anti-tumor drugs is still a challenging task.DACE arrested the cell cycle of lung epithelial cells at the G2/M phase and induced cell apoptosis by interfering with EGFR activation and its downstream signaling, including AKT, ERK, and STAT3.Taken together, these findings suggest that DACE is a promising lead compound for the development of an anti-lung-cancer drug.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmaceutical Sciences, Universidade Federal de Santa Catarina, Florianópolis, SC, Brazil; Institute of Molecular Virology (IMV), Center of Molecular Biology of Inflammation (ZMBE), Westfaelische-Wilhelms-University, Muenster, Germany.

ABSTRACT
Lung cancer is the most deadly type of cancer in humans, with non-small-cell lung cancer (NSCLC) being the most frequent and aggressive type of lung cancer showing high resistance to radiation and chemotherapy. Despite the outstanding progress made in anti-tumor therapy, discovering effective anti-tumor drugs is still a challenging task. Here we describe a new semisynthetic derivative of cucurbitacin B (DACE) as a potent inhibitor of NSCLC cell proliferation. DACE arrested the cell cycle of lung epithelial cells at the G2/M phase and induced cell apoptosis by interfering with EGFR activation and its downstream signaling, including AKT, ERK, and STAT3. Consistent with our in vitro studies, intraperitoneal application of DACE significantly suppressed the growth of mouse NSCLC that arises from type II alveolar pneumocytes due to constitutive expression of a human oncogenic c-RAF kinase (c-RAF-1-BxB) transgene in these cells. Taken together, these findings suggest that DACE is a promising lead compound for the development of an anti-lung-cancer drug.

Show MeSH
Related in: MedlinePlus