Limits...
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: 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.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.

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

Production of secreted vascular endothelial growth factor (VEGF) of OVCAR-5 (a, b, c, and d) and SKOV-3 cells (e and f) in 2D cell monolayers (black bar) and 3D ECM (grey bar). 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. The statistical difference of between 2D cell monolayers and 3D ECM are donated ** (P < 0.05, student’s t-test). Data was obtained from at least four independent experiments with triplicate
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Fig7: Production of secreted vascular endothelial growth factor (VEGF) of OVCAR-5 (a, b, c, and d) and SKOV-3 cells (e and f) in 2D cell monolayers (black bar) and 3D ECM (grey bar). 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. The statistical difference of between 2D cell monolayers and 3D ECM are donated ** (P < 0.05, student’s t-test). Data was obtained from at least four independent experiments with triplicate

Mentions: Next, we evaluated the production of secreted VEGF in the cell media after drug treatments. Single treatment of OVCAR-5 cell monolayers with cisplatin significantly increased the secreted VEGF (1.8 ng/ml control vs 3 ng/ml cisplatin, Fig. 7a). The combination of everolimus with paclitaxel (Fig. 7c) and cisplatin (Fig. 7d) reduced the VEGF secretion in both 2D cell monolayers and collagen gels. These combinations were also reproducible in SKOV-3 cell line (Fig. 7e, f). However, in SKOV-3 line the combination of everolimus with paclitaxel and cisplatin produced a greater significant reduction in collagen gels than 2D cell monolayers. Other combinations did not change the VEGF secretion in cell monolayers and collagen gels in both cell lines (data not shown).Fig. 7


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)

Production of secreted vascular endothelial growth factor (VEGF) of OVCAR-5 (a, b, c, and d) and SKOV-3 cells (e and f) in 2D cell monolayers (black bar) and 3D ECM (grey bar). 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. The statistical difference of between 2D cell monolayers and 3D ECM are donated ** (P < 0.05, student’s t-test). 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

Fig7: Production of secreted vascular endothelial growth factor (VEGF) of OVCAR-5 (a, b, c, and d) and SKOV-3 cells (e and f) in 2D cell monolayers (black bar) and 3D ECM (grey bar). 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. The statistical difference of between 2D cell monolayers and 3D ECM are donated ** (P < 0.05, student’s t-test). Data was obtained from at least four independent experiments with triplicate
Mentions: Next, we evaluated the production of secreted VEGF in the cell media after drug treatments. Single treatment of OVCAR-5 cell monolayers with cisplatin significantly increased the secreted VEGF (1.8 ng/ml control vs 3 ng/ml cisplatin, Fig. 7a). The combination of everolimus with paclitaxel (Fig. 7c) and cisplatin (Fig. 7d) reduced the VEGF secretion in both 2D cell monolayers and collagen gels. These combinations were also reproducible in SKOV-3 cell line (Fig. 7e, f). However, in SKOV-3 line the combination of everolimus with paclitaxel and cisplatin produced a greater significant reduction in collagen gels than 2D cell monolayers. Other combinations did not change the VEGF secretion in cell monolayers and collagen gels in both cell lines (data not shown).Fig. 7

Bottom Line: 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.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.

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