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Sunitinib malate synergistically potentiates anti-tumor effect of gemcitabine in human bladder cancer cells.

Yoon CY, Lee JS, Kim BS, Jeong SJ, Hong SK, Byun SS, Lee SE - Korean J Urol (2011)

Bottom Line: Sunitinib malate (Sutent; Pfizer, New York, NY, USA) is a highly selective multi-targeted agent and has been reported to have potent anti-tumor effects against various tumors, including renal cell carcinoma and gastrointestinal stromal tumors.Alterations in cell cycle (cyclin D, B1), survival (p-Akt, t-Akt), and apoptosis (Bax, Bad) regulator expression were analyzed by Western blotting.Sunitinib suppressed the expression of cyclin B1, p-Akt, and t-Akt while augmenting the expression of cyclin D and pro-apoptotic Bax and Bad in HTB5 cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, Bundang Hospital, Seoul National University College of Medicine, Seongnam, Korea.

ABSTRACT

Purpose: Sunitinib malate (Sutent; Pfizer, New York, NY, USA) is a highly selective multi-targeted agent and has been reported to have potent anti-tumor effects against various tumors, including renal cell carcinoma and gastrointestinal stromal tumors. In this study, we explored in vitro the anti-tumor effect and related molecular mechanisms of sunitinib malate against human bladder cancer cell lines. We also determined the synergistic anti-tumor effect between sunitinib and conventional cytotoxic drugs, cisplatin and gemcitabine, in bladder cancer cells.

Materials and methods: Six human cancer cell lines (HTB5, HTB9, T24, UMUC14, SW1710, and J82) were exposed to an escalating dose of sunitinib alone or in combination with cisplatin/gemcitabine, and the cytotoxic effect of the drugs was examined by CCK-8 assay. The synergistic effect between sunitinib and cisplatin/gemcitabine was determined by the combination index (CI) and clonogenic assay. Alterations in cell cycle (cyclin D, B1), survival (p-Akt, t-Akt), and apoptosis (Bax, Bad) regulator expression were analyzed by Western blotting.

Results: Like cisplatin and gemcitabine, sunitinib exerted a dose- and time-dependent anti-tumor effect in bladder cancer cells. However, sunitinib exhibited entirely different sensitivity profiles from cisplatin and gemcitabine. Sunitinib suppressed the expression of cyclin B1, p-Akt, and t-Akt while augmenting the expression of cyclin D and pro-apoptotic Bax and Bad in HTB5 cells. Analysis of the drug combination by the isobolic method and clonogenic assay revealed that sunitinib acts in synergy with gemcitabine in HTB5 cells.

Conclusions: These results indicate that sunitinib malate has a potent anti-tumor effect and may synergistically enhance the anti-tumor effect of gemcitabine in human bladder cancer cells.

No MeSH data available.


Related in: MedlinePlus

Combination treatment of human bladder cancer cells with sunitinib malate. To check the synergistic anti-tumor effect between sunitinib malate and a conventional chemotherapy agent, the HTB5 cell line was exposed to increasing doses of sunitinib malate alone or in combination with gemcitabine (A) or cisplatin (B) at a fixed ratio (1:1) for 48 hours and the anti-tumor effect was analyzed by CCK-8 assay. The median-effect plot (C) and the dose-effect plot (E) of the sunitinib and gemcitabine combination showed a synergistic anti-tumor effect for the mid-range of dose combinations, whereas the sunitinib and cisplatin combination (D, F) showed no significant synergistic effect over the whole-dose combinations tested.
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Figure 3: Combination treatment of human bladder cancer cells with sunitinib malate. To check the synergistic anti-tumor effect between sunitinib malate and a conventional chemotherapy agent, the HTB5 cell line was exposed to increasing doses of sunitinib malate alone or in combination with gemcitabine (A) or cisplatin (B) at a fixed ratio (1:1) for 48 hours and the anti-tumor effect was analyzed by CCK-8 assay. The median-effect plot (C) and the dose-effect plot (E) of the sunitinib and gemcitabine combination showed a synergistic anti-tumor effect for the mid-range of dose combinations, whereas the sunitinib and cisplatin combination (D, F) showed no significant synergistic effect over the whole-dose combinations tested.

Mentions: To test the synergistic anti-tumor effect, poorly differentiated HTB5 cells were exposed to sunitinib alone or in combination with gemcitabine. Over a wide range of concentrations, sunitinib exerted a dose-dependent anti-tumor effect and its combination with gemcitabine yielded a significantly greater anti-tumor effect than that of either agent alone (Fig. 3). These results were also confirmed by the clonogenic assay in which combination treatment of HTB5 cells with sunitinib and gemcitabine showed significantly higher suppression of colony formation (4.5±2.9% of untreated control) than single treatment with either 2.5 µM of sunitinib malate (85.3±1.31% of untreated control, p=0.005) or 0.625 µM of gemcitabine (88.1±13.6% of control, p=0.001) (Fig. 4). Because the sunitinib and gemcitabine combination indicated synergism, we performed a fixed ratio (1 to 1) combination treatment of HTB5 cells to determine the combination index. These results are summarized in Table 2 and 3. The median effect doses (Dm=IC50) for sunitinib, gemcitabine, and their combination were 6.089 µM, 198.635 µM, and 4.522 µM, respectively, indicating a more effective anti-tumor effect of the combination than of either agent alone. The median effect analysis showed CIs<1 between fa=0.15 and 0.7, which suggests a synergistic anti-tumor effect between the two drugs across the broad range of fraction affected (15-70% cell death) (Fig. 5). The dose-reduction index (DRI) also showed that the sunitinib and gemcitabine combination produced favorable dose reduction along the wide fa range (Table 3).


Sunitinib malate synergistically potentiates anti-tumor effect of gemcitabine in human bladder cancer cells.

Yoon CY, Lee JS, Kim BS, Jeong SJ, Hong SK, Byun SS, Lee SE - Korean J Urol (2011)

Combination treatment of human bladder cancer cells with sunitinib malate. To check the synergistic anti-tumor effect between sunitinib malate and a conventional chemotherapy agent, the HTB5 cell line was exposed to increasing doses of sunitinib malate alone or in combination with gemcitabine (A) or cisplatin (B) at a fixed ratio (1:1) for 48 hours and the anti-tumor effect was analyzed by CCK-8 assay. The median-effect plot (C) and the dose-effect plot (E) of the sunitinib and gemcitabine combination showed a synergistic anti-tumor effect for the mid-range of dose combinations, whereas the sunitinib and cisplatin combination (D, F) showed no significant synergistic effect over the whole-dose combinations tested.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC3037508&req=5

Figure 3: Combination treatment of human bladder cancer cells with sunitinib malate. To check the synergistic anti-tumor effect between sunitinib malate and a conventional chemotherapy agent, the HTB5 cell line was exposed to increasing doses of sunitinib malate alone or in combination with gemcitabine (A) or cisplatin (B) at a fixed ratio (1:1) for 48 hours and the anti-tumor effect was analyzed by CCK-8 assay. The median-effect plot (C) and the dose-effect plot (E) of the sunitinib and gemcitabine combination showed a synergistic anti-tumor effect for the mid-range of dose combinations, whereas the sunitinib and cisplatin combination (D, F) showed no significant synergistic effect over the whole-dose combinations tested.
Mentions: To test the synergistic anti-tumor effect, poorly differentiated HTB5 cells were exposed to sunitinib alone or in combination with gemcitabine. Over a wide range of concentrations, sunitinib exerted a dose-dependent anti-tumor effect and its combination with gemcitabine yielded a significantly greater anti-tumor effect than that of either agent alone (Fig. 3). These results were also confirmed by the clonogenic assay in which combination treatment of HTB5 cells with sunitinib and gemcitabine showed significantly higher suppression of colony formation (4.5±2.9% of untreated control) than single treatment with either 2.5 µM of sunitinib malate (85.3±1.31% of untreated control, p=0.005) or 0.625 µM of gemcitabine (88.1±13.6% of control, p=0.001) (Fig. 4). Because the sunitinib and gemcitabine combination indicated synergism, we performed a fixed ratio (1 to 1) combination treatment of HTB5 cells to determine the combination index. These results are summarized in Table 2 and 3. The median effect doses (Dm=IC50) for sunitinib, gemcitabine, and their combination were 6.089 µM, 198.635 µM, and 4.522 µM, respectively, indicating a more effective anti-tumor effect of the combination than of either agent alone. The median effect analysis showed CIs<1 between fa=0.15 and 0.7, which suggests a synergistic anti-tumor effect between the two drugs across the broad range of fraction affected (15-70% cell death) (Fig. 5). The dose-reduction index (DRI) also showed that the sunitinib and gemcitabine combination produced favorable dose reduction along the wide fa range (Table 3).

Bottom Line: Sunitinib malate (Sutent; Pfizer, New York, NY, USA) is a highly selective multi-targeted agent and has been reported to have potent anti-tumor effects against various tumors, including renal cell carcinoma and gastrointestinal stromal tumors.Alterations in cell cycle (cyclin D, B1), survival (p-Akt, t-Akt), and apoptosis (Bax, Bad) regulator expression were analyzed by Western blotting.Sunitinib suppressed the expression of cyclin B1, p-Akt, and t-Akt while augmenting the expression of cyclin D and pro-apoptotic Bax and Bad in HTB5 cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, Bundang Hospital, Seoul National University College of Medicine, Seongnam, Korea.

ABSTRACT

Purpose: Sunitinib malate (Sutent; Pfizer, New York, NY, USA) is a highly selective multi-targeted agent and has been reported to have potent anti-tumor effects against various tumors, including renal cell carcinoma and gastrointestinal stromal tumors. In this study, we explored in vitro the anti-tumor effect and related molecular mechanisms of sunitinib malate against human bladder cancer cell lines. We also determined the synergistic anti-tumor effect between sunitinib and conventional cytotoxic drugs, cisplatin and gemcitabine, in bladder cancer cells.

Materials and methods: Six human cancer cell lines (HTB5, HTB9, T24, UMUC14, SW1710, and J82) were exposed to an escalating dose of sunitinib alone or in combination with cisplatin/gemcitabine, and the cytotoxic effect of the drugs was examined by CCK-8 assay. The synergistic effect between sunitinib and cisplatin/gemcitabine was determined by the combination index (CI) and clonogenic assay. Alterations in cell cycle (cyclin D, B1), survival (p-Akt, t-Akt), and apoptosis (Bax, Bad) regulator expression were analyzed by Western blotting.

Results: Like cisplatin and gemcitabine, sunitinib exerted a dose- and time-dependent anti-tumor effect in bladder cancer cells. However, sunitinib exhibited entirely different sensitivity profiles from cisplatin and gemcitabine. Sunitinib suppressed the expression of cyclin B1, p-Akt, and t-Akt while augmenting the expression of cyclin D and pro-apoptotic Bax and Bad in HTB5 cells. Analysis of the drug combination by the isobolic method and clonogenic assay revealed that sunitinib acts in synergy with gemcitabine in HTB5 cells.

Conclusions: These results indicate that sunitinib malate has a potent anti-tumor effect and may synergistically enhance the anti-tumor effect of gemcitabine in human bladder cancer cells.

No MeSH data available.


Related in: MedlinePlus