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NCX-4040, a nitric oxide-releasing aspirin, sensitizes drug-resistant human ovarian xenograft tumors to cisplatin by depletion of cellular thiols.

Bratasz A, Selvendiran K, Wasowicz T, Bobko A, Khramtsov VV, Ignarro LJ, Kuppusamy P - J Transl Med (2008)

Bottom Line: Cells treated with NCX-4040 (25 microM) showed a significant reduction of cell viability (A2780 WT, 34.9 +/- 8.7%; A2780 cDDP, 41.7 +/- 7.6%; p < 0.05).EPR imaging of tissue redox and thiol measurements showed a 5.5-fold reduction (p < 0.01) of glutathione in NCX-4040-treated A2780 cDDP tumors when compared to untreated controls.The results suggested that NCX-4040 could resensitize drug-resistant ovarian cancer cells to cisplatin possibly by depletion of cellular thiols.

View Article: PubMed Central - HTML - PubMed

Affiliation: Center for Biomedical EPR Spectroscopy and Imaging, Davis Heart and Lung Research Institute, Department of Internal Medicine, The Ohio State University, Columbus, OH 43210, USA. kuppusamy.1@osu.edu

ABSTRACT

Background: Ovarian carcinoma is the leading cause of mortality among gynecological cancers in the world. The high mortality rate is associated with lack of early diagnosis and development of drug resistance. The antitumor efficacy and mechanism of NCX-4040, a nitric oxide-releasing aspirin derivative, against ovarian cancer is studied.

Methods: NCX-4040, alone or in combination with cisplatin (cis-diamminedichloroplatinum, cDDP), was studied in cisplatin-sensitive (A2780 WT) and cisplatin-resistant (A2780 cDDP) cell lines as well as xenograft tumors grown in nude mice. Electron paramagnetic resonance (EPR) was used for measurements of nitric oxide and redox state. Immunoblotting analysis of A2780 cDDP tumor xenografts from mice was used for mechanistic studies.

Results: Cells treated with NCX-4040 (25 microM) showed a significant reduction of cell viability (A2780 WT, 34.9 +/- 8.7%; A2780 cDDP, 41.7 +/- 7.6%; p < 0.05). Further, NCX-4040 significantly enhanced the sensitivity of A2780 cDDP cells (cisplatin alone, 80.6 +/- 11.8% versus NCX-4040+cisplatin, 26.4 +/- 7.6%; p < 0.01) and xenograft tumors (cisplatin alone, 74.0 +/- 4.4% versus NCX-4040+cisplatin, 56.4 +/- 7.8%; p < 0.05), to cisplatin treatment. EPR imaging of tissue redox and thiol measurements showed a 5.5-fold reduction (p < 0.01) of glutathione in NCX-4040-treated A2780 cDDP tumors when compared to untreated controls. Immunoblotting analysis of A2780 cDDP tumor xenografts from mice treated with NCX-4040 and cisplatin revealed significant downregulation of pEGFR (Tyr845 and Tyr992) and pSTAT3 (Tyr705 and Ser727) expression.

Conclusion: The results suggested that NCX-4040 could resensitize drug-resistant ovarian cancer cells to cisplatin possibly by depletion of cellular thiols. Thus NCX-4040 appears to be a potential therapeutic agent for the treatment of human ovarian carcinoma and cisplatin-resistant malignancies.

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Effect of NCX-4040 and/or cisplatin (cDDP) on the growth volume of human ovarian cancer xenograft tumor in mice. Groups of mice (4–6 per group) were inoculated with the A2780 cDDP human ovarian cancer cells in the upper portion of hind leg. Seven days after inoculation, two groups were injected daily (i.p.) with NCX-4040 (5 mg/kg), of which one group received a single i.p. injection of cisplatin on day 11 (8 mg/kg). The third group received single dose of cisplatin on day 11 (8 mg/kg). One group received aspirin (5 mg/kg, daily). Control group received vehicle. (A) Tumor growth curve for control, cisplatin, and combination of treatment (NCX-4040 and cisplatin). (B) Tumor growth volume data (mean ± SE, expressed as percent of control group) on the 19th day after injection of cancer cells. *p < 0.05 compared to control group. **p < 0.05 versus cDDP group. NCX-4040 and aspirin (ASA), a metabolic product of NCX-4040, showed no significant effect. The results show that pretreatment with NCX-4040 was effective in enhancing the efficacy of cisplatin in inhibiting the growth of cisplatin-resistant ovarian cancer xenografts in mice.
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Figure 4: Effect of NCX-4040 and/or cisplatin (cDDP) on the growth volume of human ovarian cancer xenograft tumor in mice. Groups of mice (4–6 per group) were inoculated with the A2780 cDDP human ovarian cancer cells in the upper portion of hind leg. Seven days after inoculation, two groups were injected daily (i.p.) with NCX-4040 (5 mg/kg), of which one group received a single i.p. injection of cisplatin on day 11 (8 mg/kg). The third group received single dose of cisplatin on day 11 (8 mg/kg). One group received aspirin (5 mg/kg, daily). Control group received vehicle. (A) Tumor growth curve for control, cisplatin, and combination of treatment (NCX-4040 and cisplatin). (B) Tumor growth volume data (mean ± SE, expressed as percent of control group) on the 19th day after injection of cancer cells. *p < 0.05 compared to control group. **p < 0.05 versus cDDP group. NCX-4040 and aspirin (ASA), a metabolic product of NCX-4040, showed no significant effect. The results show that pretreatment with NCX-4040 was effective in enhancing the efficacy of cisplatin in inhibiting the growth of cisplatin-resistant ovarian cancer xenografts in mice.

Mentions: The anti-tumor efficacy of NCX-4040 was studied using A2780 cDDP tumor xenografts in mice. Tumor-bearing animals, on day 7 post-inoculation, were treated with NCX-4040 and/or cisplatin, as described in the Methods section. The tumor volumes were measured every 2 days for 19 days post-inoculation. As shown in Figure 4, NCX-4040 treatment alone did not show any significant reduction in tumor volume (85.8 ± 9.8% versus Control on day 19; Figure 4B). Cisplatin (cDDP) treatment alone significantly reduced the tumor growth volume (74.0 ± 4.4% versus Control). However, animals treated with a combination of cisplatin and NCX-4040 showed a significant reduction in tumor volume visible from second day after cisplatin treatment and reached 56.4 ± 7.8% versus Control on day 19. Aspirin (ASA), a metabolic product of NCX-4040, did not show any significant effect on tumor growth volume (Figure 4B). The results clearly demonstrated that NCX-4040, in combination with cisplatin, was more effective than cisplatin alone in inhibiting the growth of cisplatin-resistant ovarian cancer xenografts in mice.


NCX-4040, a nitric oxide-releasing aspirin, sensitizes drug-resistant human ovarian xenograft tumors to cisplatin by depletion of cellular thiols.

Bratasz A, Selvendiran K, Wasowicz T, Bobko A, Khramtsov VV, Ignarro LJ, Kuppusamy P - J Transl Med (2008)

Effect of NCX-4040 and/or cisplatin (cDDP) on the growth volume of human ovarian cancer xenograft tumor in mice. Groups of mice (4–6 per group) were inoculated with the A2780 cDDP human ovarian cancer cells in the upper portion of hind leg. Seven days after inoculation, two groups were injected daily (i.p.) with NCX-4040 (5 mg/kg), of which one group received a single i.p. injection of cisplatin on day 11 (8 mg/kg). The third group received single dose of cisplatin on day 11 (8 mg/kg). One group received aspirin (5 mg/kg, daily). Control group received vehicle. (A) Tumor growth curve for control, cisplatin, and combination of treatment (NCX-4040 and cisplatin). (B) Tumor growth volume data (mean ± SE, expressed as percent of control group) on the 19th day after injection of cancer cells. *p < 0.05 compared to control group. **p < 0.05 versus cDDP group. NCX-4040 and aspirin (ASA), a metabolic product of NCX-4040, showed no significant effect. The results show that pretreatment with NCX-4040 was effective in enhancing the efficacy of cisplatin in inhibiting the growth of cisplatin-resistant ovarian cancer xenografts in mice.
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Figure 4: Effect of NCX-4040 and/or cisplatin (cDDP) on the growth volume of human ovarian cancer xenograft tumor in mice. Groups of mice (4–6 per group) were inoculated with the A2780 cDDP human ovarian cancer cells in the upper portion of hind leg. Seven days after inoculation, two groups were injected daily (i.p.) with NCX-4040 (5 mg/kg), of which one group received a single i.p. injection of cisplatin on day 11 (8 mg/kg). The third group received single dose of cisplatin on day 11 (8 mg/kg). One group received aspirin (5 mg/kg, daily). Control group received vehicle. (A) Tumor growth curve for control, cisplatin, and combination of treatment (NCX-4040 and cisplatin). (B) Tumor growth volume data (mean ± SE, expressed as percent of control group) on the 19th day after injection of cancer cells. *p < 0.05 compared to control group. **p < 0.05 versus cDDP group. NCX-4040 and aspirin (ASA), a metabolic product of NCX-4040, showed no significant effect. The results show that pretreatment with NCX-4040 was effective in enhancing the efficacy of cisplatin in inhibiting the growth of cisplatin-resistant ovarian cancer xenografts in mice.
Mentions: The anti-tumor efficacy of NCX-4040 was studied using A2780 cDDP tumor xenografts in mice. Tumor-bearing animals, on day 7 post-inoculation, were treated with NCX-4040 and/or cisplatin, as described in the Methods section. The tumor volumes were measured every 2 days for 19 days post-inoculation. As shown in Figure 4, NCX-4040 treatment alone did not show any significant reduction in tumor volume (85.8 ± 9.8% versus Control on day 19; Figure 4B). Cisplatin (cDDP) treatment alone significantly reduced the tumor growth volume (74.0 ± 4.4% versus Control). However, animals treated with a combination of cisplatin and NCX-4040 showed a significant reduction in tumor volume visible from second day after cisplatin treatment and reached 56.4 ± 7.8% versus Control on day 19. Aspirin (ASA), a metabolic product of NCX-4040, did not show any significant effect on tumor growth volume (Figure 4B). The results clearly demonstrated that NCX-4040, in combination with cisplatin, was more effective than cisplatin alone in inhibiting the growth of cisplatin-resistant ovarian cancer xenografts in mice.

Bottom Line: Cells treated with NCX-4040 (25 microM) showed a significant reduction of cell viability (A2780 WT, 34.9 +/- 8.7%; A2780 cDDP, 41.7 +/- 7.6%; p < 0.05).EPR imaging of tissue redox and thiol measurements showed a 5.5-fold reduction (p < 0.01) of glutathione in NCX-4040-treated A2780 cDDP tumors when compared to untreated controls.The results suggested that NCX-4040 could resensitize drug-resistant ovarian cancer cells to cisplatin possibly by depletion of cellular thiols.

View Article: PubMed Central - HTML - PubMed

Affiliation: Center for Biomedical EPR Spectroscopy and Imaging, Davis Heart and Lung Research Institute, Department of Internal Medicine, The Ohio State University, Columbus, OH 43210, USA. kuppusamy.1@osu.edu

ABSTRACT

Background: Ovarian carcinoma is the leading cause of mortality among gynecological cancers in the world. The high mortality rate is associated with lack of early diagnosis and development of drug resistance. The antitumor efficacy and mechanism of NCX-4040, a nitric oxide-releasing aspirin derivative, against ovarian cancer is studied.

Methods: NCX-4040, alone or in combination with cisplatin (cis-diamminedichloroplatinum, cDDP), was studied in cisplatin-sensitive (A2780 WT) and cisplatin-resistant (A2780 cDDP) cell lines as well as xenograft tumors grown in nude mice. Electron paramagnetic resonance (EPR) was used for measurements of nitric oxide and redox state. Immunoblotting analysis of A2780 cDDP tumor xenografts from mice was used for mechanistic studies.

Results: Cells treated with NCX-4040 (25 microM) showed a significant reduction of cell viability (A2780 WT, 34.9 +/- 8.7%; A2780 cDDP, 41.7 +/- 7.6%; p < 0.05). Further, NCX-4040 significantly enhanced the sensitivity of A2780 cDDP cells (cisplatin alone, 80.6 +/- 11.8% versus NCX-4040+cisplatin, 26.4 +/- 7.6%; p < 0.01) and xenograft tumors (cisplatin alone, 74.0 +/- 4.4% versus NCX-4040+cisplatin, 56.4 +/- 7.8%; p < 0.05), to cisplatin treatment. EPR imaging of tissue redox and thiol measurements showed a 5.5-fold reduction (p < 0.01) of glutathione in NCX-4040-treated A2780 cDDP tumors when compared to untreated controls. Immunoblotting analysis of A2780 cDDP tumor xenografts from mice treated with NCX-4040 and cisplatin revealed significant downregulation of pEGFR (Tyr845 and Tyr992) and pSTAT3 (Tyr705 and Ser727) expression.

Conclusion: The results suggested that NCX-4040 could resensitize drug-resistant ovarian cancer cells to cisplatin possibly by depletion of cellular thiols. Thus NCX-4040 appears to be a potential therapeutic agent for the treatment of human ovarian carcinoma and cisplatin-resistant malignancies.

Show MeSH
Related in: MedlinePlus