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A method to investigate the anti-metabolic activity of anti-cancer agents on ovarian cancer cells cultured in a 96-well high throughput format.

Hogg SJ, Evans JJ, Sykes PH, Chitcholtan K - J Ovarian Res (2015)

Bottom Line: This surface is rich in extracellular matrix (ECM) proteins, which have profound effects on cellular behaviour and can facilitate cancer progression.The primary endpoints were to measure the reduction of cellular metabolism and secretion of vascular endothelial growth factor (VEGF).The responses to drugs were prominently observed in collagen gels, but they had little effect on 2D cell monolayers.

View Article: PubMed Central - PubMed

Affiliation: Peter MacCallum Cancer Centre, St Andrews Place, East Melbourne, 3002, VIC, Australia. Simon.Hogg@patermac.org.

ABSTRACT

Background: An early step of advanced ovarian cancer begins when floating cancerous cells as single cells or small clusters grow on the peritoneal surface. This surface is rich in extracellular matrix (ECM) proteins, which have profound effects on cellular behaviour and can facilitate cancer progression. Subsequently, this ECM may alter cellular metabolism making cancer cells susceptible to chemotherapeutic agents differently. Therefore, generating a cell culture tool in vitro that includes the interaction between ECM and cancer cells will facilitate our understanding of how cancer cells behave during cancer treatment. There is some evidence to suggest that in an in vitro model that includes ECM components such as collagens will provide a better predictive tool for drug evaluation than a traditional cell monolayer (2D) culture model.

Findings: As a proof -of- concept, we made a collagen gel in a 96-well plate format and utilised this to evaluate the efficacy of clinical cytotoxic drugs, a targeted drug, and food compounds in single and combination treatments. The primary endpoints were to measure the reduction of cellular metabolism and secretion of vascular endothelial growth factor (VEGF). The invasive capacity of cancer cells was observed in collagen gels and it was cell line-dependent. The responses to drugs were prominently observed in collagen gels, but they had little effect on 2D cell monolayers. These responses were cell line- and type of drug-dependent.

Conclusions: The collagen gel in a 96 well plate format was easy to set up and could have potential to identify drug sensitivity in the clinical management of women with platinum resistant ovarian cancer.

No MeSH data available.


Related in: MedlinePlus

Cellular metabolism profiles of OVCAR-5 cell line with single and combination treatment of EGCG + paclitaxel (a), EGCG + cisplatin (b), EGCG + everolimus (c), paclitaxel + cisplatin (d) paclitaxel + everolimus (e) and cisplatin + everolimus (f) in 2D cell monolayers (black bar) and 3D ECM (grey bar). The representative graph in 2D cell monolayers and 3D ECM was the relative value to the control. The statistical difference of single and combination in 2D cell monolayers (*P < 0.05, student’s t-test) and 3D ECM (#P < 0.05, student’s t-test) was compared between the control and treated cells. Data was obtained from at least four independent experiments with triplicate
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Fig4: Cellular metabolism profiles of OVCAR-5 cell line with single and combination treatment of EGCG + paclitaxel (a), EGCG + cisplatin (b), EGCG + everolimus (c), paclitaxel + cisplatin (d) paclitaxel + everolimus (e) and cisplatin + everolimus (f) in 2D cell monolayers (black bar) and 3D ECM (grey bar). The representative graph in 2D cell monolayers and 3D ECM was the relative value to the control. The statistical difference of single and combination in 2D cell monolayers (*P < 0.05, student’s t-test) and 3D ECM (#P < 0.05, student’s t-test) was compared between the control and treated cells. Data was obtained from at least four independent experiments with triplicate

Mentions: Next, we determined the efficacy of single drug and combination treatments in cell monolayers and cells on top of collagen gels. Single and combination treatments were selectively affected in OVCAR-5 cells in collagen gel (Figs. 3 and 4). Single treatments that reduced cellular metabolism only in collagen gel compared to the control included resveratrol (29 %, Fig. 3a), EGCG (35 %, Fig. 3a), and paclitaxel (28.1 %, Fig. 3b). Everolimus (Fig. 3d) reduced cellular metabolism in both cell monolayers (13 %) and collagen gel (16 %). The cell monolayers of OVCAR-5 cell line did not show any marked reduction of cellular metabolism except cisplatin (12 %, Fig. 3c).Fig. 3


A method to investigate the anti-metabolic activity of anti-cancer agents on ovarian cancer cells cultured in a 96-well high throughput format.

Hogg SJ, Evans JJ, Sykes PH, Chitcholtan K - J Ovarian Res (2015)

Cellular metabolism profiles of OVCAR-5 cell line with single and combination treatment of EGCG + paclitaxel (a), EGCG + cisplatin (b), EGCG + everolimus (c), paclitaxel + cisplatin (d) paclitaxel + everolimus (e) and cisplatin + everolimus (f) in 2D cell monolayers (black bar) and 3D ECM (grey bar). The representative graph in 2D cell monolayers and 3D ECM was the relative value to the control. The statistical difference of single and combination in 2D cell monolayers (*P < 0.05, student’s t-test) and 3D ECM (#P < 0.05, student’s t-test) was compared between the control and treated cells. Data was obtained from at least four independent experiments with triplicate
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4491427&req=5

Fig4: Cellular metabolism profiles of OVCAR-5 cell line with single and combination treatment of EGCG + paclitaxel (a), EGCG + cisplatin (b), EGCG + everolimus (c), paclitaxel + cisplatin (d) paclitaxel + everolimus (e) and cisplatin + everolimus (f) in 2D cell monolayers (black bar) and 3D ECM (grey bar). The representative graph in 2D cell monolayers and 3D ECM was the relative value to the control. The statistical difference of single and combination in 2D cell monolayers (*P < 0.05, student’s t-test) and 3D ECM (#P < 0.05, student’s t-test) was compared between the control and treated cells. Data was obtained from at least four independent experiments with triplicate
Mentions: Next, we determined the efficacy of single drug and combination treatments in cell monolayers and cells on top of collagen gels. Single and combination treatments were selectively affected in OVCAR-5 cells in collagen gel (Figs. 3 and 4). Single treatments that reduced cellular metabolism only in collagen gel compared to the control included resveratrol (29 %, Fig. 3a), EGCG (35 %, Fig. 3a), and paclitaxel (28.1 %, Fig. 3b). Everolimus (Fig. 3d) reduced cellular metabolism in both cell monolayers (13 %) and collagen gel (16 %). The cell monolayers of OVCAR-5 cell line did not show any marked reduction of cellular metabolism except cisplatin (12 %, Fig. 3c).Fig. 3

Bottom Line: This surface is rich in extracellular matrix (ECM) proteins, which have profound effects on cellular behaviour and can facilitate cancer progression.The primary endpoints were to measure the reduction of cellular metabolism and secretion of vascular endothelial growth factor (VEGF).The responses to drugs were prominently observed in collagen gels, but they had little effect on 2D cell monolayers.

View Article: PubMed Central - PubMed

Affiliation: Peter MacCallum Cancer Centre, St Andrews Place, East Melbourne, 3002, VIC, Australia. Simon.Hogg@patermac.org.

ABSTRACT

Background: An early step of advanced ovarian cancer begins when floating cancerous cells as single cells or small clusters grow on the peritoneal surface. This surface is rich in extracellular matrix (ECM) proteins, which have profound effects on cellular behaviour and can facilitate cancer progression. Subsequently, this ECM may alter cellular metabolism making cancer cells susceptible to chemotherapeutic agents differently. Therefore, generating a cell culture tool in vitro that includes the interaction between ECM and cancer cells will facilitate our understanding of how cancer cells behave during cancer treatment. There is some evidence to suggest that in an in vitro model that includes ECM components such as collagens will provide a better predictive tool for drug evaluation than a traditional cell monolayer (2D) culture model.

Findings: As a proof -of- concept, we made a collagen gel in a 96-well plate format and utilised this to evaluate the efficacy of clinical cytotoxic drugs, a targeted drug, and food compounds in single and combination treatments. The primary endpoints were to measure the reduction of cellular metabolism and secretion of vascular endothelial growth factor (VEGF). The invasive capacity of cancer cells was observed in collagen gels and it was cell line-dependent. The responses to drugs were prominently observed in collagen gels, but they had little effect on 2D cell monolayers. These responses were cell line- and type of drug-dependent.

Conclusions: The collagen gel in a 96 well plate format was easy to set up and could have potential to identify drug sensitivity in the clinical management of women with platinum resistant ovarian cancer.

No MeSH data available.


Related in: MedlinePlus