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ZNF93 increases resistance to ET-743 (Trabectedin; Yondelis) and PM00104 (Zalypsis) in human cancer cell lines.

Duan Z, Choy E, Harmon D, Yang C, Ryu K, Schwab J, Mankin H, Hornicek FJ - PLoS ONE (2009)

Bottom Line: We found that a large number of genes have altered expression levels in CS-1/ER and CS-1/PR when compared to the parental cell line. 595 CS-1/ER and 498 CS-1/PR genes were identified as overexpressing; 856 CS-1/ER and 874 CS-1/PR transcripts were identified as underexpressing.These genes have not been previously associated with drug resistance in tumor cells.This study suggests that zinc finger proteins, and ZNF93 in particular, are involved in resistance to ET-743 and PM00104.

View Article: PubMed Central - PubMed

Affiliation: Sarcoma Biology Laboratory, Center for Sarcoma and Connective Tissue Oncology, Massachusetts General Hospital, Boston, Massachusetts, United States of America.

ABSTRACT

Background: ET-743 (trabectedin, Yondelis) and PM00104 (Zalypsis) are marine derived compounds that have antitumor activity. ET-743 and PM00104 exposure over sustained periods of treatment will result in the development of drug resistance, but the mechanisms which lead to resistance are not yet understood.

Methodology/principal findings: Human chondrosarcoma cell lines resistant to ET-743 (CS-1/ER) or PM00104 (CS-1/PR) were established in this study. The CS-1/ER and CS-1/PR exhibited cross resistance to cisplatin and methotrexate but not to doxorubicin. Human Affymetrix Gene Chip arrays were used to examine relative gene expression in these cell lines. We found that a large number of genes have altered expression levels in CS-1/ER and CS-1/PR when compared to the parental cell line. 595 CS-1/ER and 498 CS-1/PR genes were identified as overexpressing; 856 CS-1/ER and 874 CS-1/PR transcripts were identified as underexpressing. Three zinc finger protein (ZNF) genes were on the top 10 overexpressed genes list. These genes have not been previously associated with drug resistance in tumor cells. Differential expressions of ZNF93 and ZNF43 genes were confirmed in both CS-1/ER and CS-1/PR resistant cell lines by real-time RT-PCR. ZNF93 was overexpressed in two ET-743 resistant Ewing sarcoma cell lines as well as in a cisplatin resistant ovarian cancer cell line, but was not overexpressed in paclitaxel resistant cell lines. ZNF93 knockdown by siRNA in CS-1/ER and CS-1/PR caused increased sensitivity for ET-743, PM00104, and cisplatin. Furthermore, ZNF93 transfected CS-1 cells are relatively resistant to ET-743, PM00104 and cisplatin.

Conclusions/significance: This study suggests that zinc finger proteins, and ZNF93 in particular, are involved in resistance to ET-743 and PM00104.

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Related in: MedlinePlus

Exogenous expression of ZNF93 confers PM00104, ET743 resistance.A, B and C: Relative cytotoxicity of ET-743,PM00104 and cisplatin in CS-1 derived cell lines (CS-1/pIRESZNF93) stably transfected with a pIRESZNF93 expression vector and in the parental (CS-1) and empty vector (CS-1/pIRES) controls were assessed using the MTT assay. All samples were analyzed in triplicate. D: Confirmation of ZNF93 overexpression in pIRESZNF93 tranfected CS-1 cells by real-time RT-PCR as described in Materials and Methods.
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pone-0006967-g006: Exogenous expression of ZNF93 confers PM00104, ET743 resistance.A, B and C: Relative cytotoxicity of ET-743,PM00104 and cisplatin in CS-1 derived cell lines (CS-1/pIRESZNF93) stably transfected with a pIRESZNF93 expression vector and in the parental (CS-1) and empty vector (CS-1/pIRES) controls were assessed using the MTT assay. All samples were analyzed in triplicate. D: Confirmation of ZNF93 overexpression in pIRESZNF93 tranfected CS-1 cells by real-time RT-PCR as described in Materials and Methods.

Mentions: Our analyses of endogenous ZNF93 expression demonstrated that ZNF93 is frequently upregulated in ET-743, PM00104 and cisplatin multiple drug resistant cancer cell lines, ZNF93 down-regulation by siRNA could partially recover sensitivity to ET-743,PM00104 and cispatin. These results suggest that the ZNF93 protein may be critically involved in the development of resistance to these drugs. To further determine whether ZNF93 directly participates in the establishment of the resistant phenotype, we transfected the ET-743 and PM00104 sensitive cell line CS-1 with a ZNF93 expression vector and generated stable cell line which overexpress ZNF93 (Figure 6D). We then asked if exogenous overexpression of ZNF93 was sufficient to confer increased ET-743 and PM00104 resistance. The results from the MTT assay in the tranfected cell lines are shown in Fig. 6. ZNF93 transfected CS-1 cells are relatively resistant to ET-743, PM00104 and cisplatin, whereas no significant change in resistance was observed in CS-1 cells transfected with the empty vector (Figure 6A to C). Elevated ZNF93 expression in the tranfected cells was confirmed by real-time RT-PCR (Fig. 6D).


ZNF93 increases resistance to ET-743 (Trabectedin; Yondelis) and PM00104 (Zalypsis) in human cancer cell lines.

Duan Z, Choy E, Harmon D, Yang C, Ryu K, Schwab J, Mankin H, Hornicek FJ - PLoS ONE (2009)

Exogenous expression of ZNF93 confers PM00104, ET743 resistance.A, B and C: Relative cytotoxicity of ET-743,PM00104 and cisplatin in CS-1 derived cell lines (CS-1/pIRESZNF93) stably transfected with a pIRESZNF93 expression vector and in the parental (CS-1) and empty vector (CS-1/pIRES) controls were assessed using the MTT assay. All samples were analyzed in triplicate. D: Confirmation of ZNF93 overexpression in pIRESZNF93 tranfected CS-1 cells by real-time RT-PCR as described in Materials and Methods.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2734182&req=5

pone-0006967-g006: Exogenous expression of ZNF93 confers PM00104, ET743 resistance.A, B and C: Relative cytotoxicity of ET-743,PM00104 and cisplatin in CS-1 derived cell lines (CS-1/pIRESZNF93) stably transfected with a pIRESZNF93 expression vector and in the parental (CS-1) and empty vector (CS-1/pIRES) controls were assessed using the MTT assay. All samples were analyzed in triplicate. D: Confirmation of ZNF93 overexpression in pIRESZNF93 tranfected CS-1 cells by real-time RT-PCR as described in Materials and Methods.
Mentions: Our analyses of endogenous ZNF93 expression demonstrated that ZNF93 is frequently upregulated in ET-743, PM00104 and cisplatin multiple drug resistant cancer cell lines, ZNF93 down-regulation by siRNA could partially recover sensitivity to ET-743,PM00104 and cispatin. These results suggest that the ZNF93 protein may be critically involved in the development of resistance to these drugs. To further determine whether ZNF93 directly participates in the establishment of the resistant phenotype, we transfected the ET-743 and PM00104 sensitive cell line CS-1 with a ZNF93 expression vector and generated stable cell line which overexpress ZNF93 (Figure 6D). We then asked if exogenous overexpression of ZNF93 was sufficient to confer increased ET-743 and PM00104 resistance. The results from the MTT assay in the tranfected cell lines are shown in Fig. 6. ZNF93 transfected CS-1 cells are relatively resistant to ET-743, PM00104 and cisplatin, whereas no significant change in resistance was observed in CS-1 cells transfected with the empty vector (Figure 6A to C). Elevated ZNF93 expression in the tranfected cells was confirmed by real-time RT-PCR (Fig. 6D).

Bottom Line: We found that a large number of genes have altered expression levels in CS-1/ER and CS-1/PR when compared to the parental cell line. 595 CS-1/ER and 498 CS-1/PR genes were identified as overexpressing; 856 CS-1/ER and 874 CS-1/PR transcripts were identified as underexpressing.These genes have not been previously associated with drug resistance in tumor cells.This study suggests that zinc finger proteins, and ZNF93 in particular, are involved in resistance to ET-743 and PM00104.

View Article: PubMed Central - PubMed

Affiliation: Sarcoma Biology Laboratory, Center for Sarcoma and Connective Tissue Oncology, Massachusetts General Hospital, Boston, Massachusetts, United States of America.

ABSTRACT

Background: ET-743 (trabectedin, Yondelis) and PM00104 (Zalypsis) are marine derived compounds that have antitumor activity. ET-743 and PM00104 exposure over sustained periods of treatment will result in the development of drug resistance, but the mechanisms which lead to resistance are not yet understood.

Methodology/principal findings: Human chondrosarcoma cell lines resistant to ET-743 (CS-1/ER) or PM00104 (CS-1/PR) were established in this study. The CS-1/ER and CS-1/PR exhibited cross resistance to cisplatin and methotrexate but not to doxorubicin. Human Affymetrix Gene Chip arrays were used to examine relative gene expression in these cell lines. We found that a large number of genes have altered expression levels in CS-1/ER and CS-1/PR when compared to the parental cell line. 595 CS-1/ER and 498 CS-1/PR genes were identified as overexpressing; 856 CS-1/ER and 874 CS-1/PR transcripts were identified as underexpressing. Three zinc finger protein (ZNF) genes were on the top 10 overexpressed genes list. These genes have not been previously associated with drug resistance in tumor cells. Differential expressions of ZNF93 and ZNF43 genes were confirmed in both CS-1/ER and CS-1/PR resistant cell lines by real-time RT-PCR. ZNF93 was overexpressed in two ET-743 resistant Ewing sarcoma cell lines as well as in a cisplatin resistant ovarian cancer cell line, but was not overexpressed in paclitaxel resistant cell lines. ZNF93 knockdown by siRNA in CS-1/ER and CS-1/PR caused increased sensitivity for ET-743, PM00104, and cisplatin. Furthermore, ZNF93 transfected CS-1 cells are relatively resistant to ET-743, PM00104 and cisplatin.

Conclusions/significance: This study suggests that zinc finger proteins, and ZNF93 in particular, are involved in resistance to ET-743 and PM00104.

Show MeSH
Related in: MedlinePlus