Limits...
WHI-P154 enhances the chemotherapeutic effect of anticancer agents in ABCG2-overexpressing cells.

Zhang H, Zhang YK, Wang YJ, Kathawala RJ, Patel A, Zhu H, Sodani K, Talele TT, Ambudkar SV, Chen ZS, Fu LW - Cancer Sci. (2014)

Bottom Line: We found that WHI-P154 significantly enhanced the sensitivity of ABCG2-overexpressing cells to its substrates.WHI-P154 moderately sensitized ABCB1-overexpressing KB-C2 cells to its substrates whereas showed no sensitizing effect on ABCC1-, ABCC2 or ABCC10-mediated drug resistance.Collectively, these findings highlighted WHI-P154 could significantly reverse ABCG2-mediated multidrug drug resistance by directly blocking the efflux function.

View Article: PubMed Central - PubMed

Affiliation: Sun Yat-Sen University Cancer Center, State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Guangzhou, China; Department of Pharmaceutical Sciences, College of Pharmacy and Health Sciences, St John's University, Queens, New York, USA.

Show MeSH

Related in: MedlinePlus

The effect of WHI-P154 on the expression levels and localization of ABCG2, and on the blockade of STAT3, AKT and ERK1/2 phosphorylation in ABCG2-overexpressing cells. Western blot showed the effect of WHI-P154 on the expression level of ABCG2 after cells were treated with 4 μM of WHI-P154 for 0, 24, 48, and 72 h (a). Grayscale ratios of BCRP/β-actin were analyzed with Image J. The differences were statistically not significant (P > 0.05) (c). Similarly, WHI-P154 did not significantly alter the expression level of ABCG2 in ABCG2-482-R2 cells (b, d). Figure 3(a, c) representative result is shown from at least three independent experiments. Immunofluorescence staining showed membranous cellular localization of ABCG2 in H460/MX20 cells after treatment with WHI-P154 for 72 h. ABCG2 staining is shown in green. Fluorescent DAPI (blue) counterstains the nuclei (e). Western blot showed the effect of various concentrations of WHI-P154 on the phosphorylation of STAT3, AKT and ERK1/2 (f). Representative results are shown and similar results were obtained in two other independent trials.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4317847&req=5

fig03: The effect of WHI-P154 on the expression levels and localization of ABCG2, and on the blockade of STAT3, AKT and ERK1/2 phosphorylation in ABCG2-overexpressing cells. Western blot showed the effect of WHI-P154 on the expression level of ABCG2 after cells were treated with 4 μM of WHI-P154 for 0, 24, 48, and 72 h (a). Grayscale ratios of BCRP/β-actin were analyzed with Image J. The differences were statistically not significant (P > 0.05) (c). Similarly, WHI-P154 did not significantly alter the expression level of ABCG2 in ABCG2-482-R2 cells (b, d). Figure 3(a, c) representative result is shown from at least three independent experiments. Immunofluorescence staining showed membranous cellular localization of ABCG2 in H460/MX20 cells after treatment with WHI-P154 for 72 h. ABCG2 staining is shown in green. Fluorescent DAPI (blue) counterstains the nuclei (e). Western blot showed the effect of various concentrations of WHI-P154 on the phosphorylation of STAT3, AKT and ERK1/2 (f). Representative results are shown and similar results were obtained in two other independent trials.

Mentions: The reversal effect of ABCG2-mediated MDR could be achieved by either inhibiting ABCG2 function or decreasing the expression levels of ABCG2 protein. Therefore, Western blot analysis was performed to further ascertain the effect of WHI-P154 on the expression levels of ABCG2. Our results showed the expression levels of ABCG2 protein (Fig. 3) were not significantly altered in H460/MX20 cells after treatment with 4 μM WHI-P154 for 0, 24, 48 and 72 h, and the data also showed low-level expression of ABCG2 in parent H460 cells (Fig. 3a, c). The grayscale ratios of ABCG2/β-actin were proportional to the ABCG2 protein levels (Fig. 3c). Similarly, WHI-P154 did not significantly alter the expression levels of ABCG2 in ABCG2-482-R2 cells (Fig. 3b, d). In addition, we performed immunofluorescence to determine the effect of WHI-P154 on the cellular localization of ABCG2. As shown in Figure 3(e), WHI-P154 did not alter the localization of ABCG2. There results indicated that neither the expression levels nor localization of ABCG2 was significantly altered by WHI-P154.


WHI-P154 enhances the chemotherapeutic effect of anticancer agents in ABCG2-overexpressing cells.

Zhang H, Zhang YK, Wang YJ, Kathawala RJ, Patel A, Zhu H, Sodani K, Talele TT, Ambudkar SV, Chen ZS, Fu LW - Cancer Sci. (2014)

The effect of WHI-P154 on the expression levels and localization of ABCG2, and on the blockade of STAT3, AKT and ERK1/2 phosphorylation in ABCG2-overexpressing cells. Western blot showed the effect of WHI-P154 on the expression level of ABCG2 after cells were treated with 4 μM of WHI-P154 for 0, 24, 48, and 72 h (a). Grayscale ratios of BCRP/β-actin were analyzed with Image J. The differences were statistically not significant (P > 0.05) (c). Similarly, WHI-P154 did not significantly alter the expression level of ABCG2 in ABCG2-482-R2 cells (b, d). Figure 3(a, c) representative result is shown from at least three independent experiments. Immunofluorescence staining showed membranous cellular localization of ABCG2 in H460/MX20 cells after treatment with WHI-P154 for 72 h. ABCG2 staining is shown in green. Fluorescent DAPI (blue) counterstains the nuclei (e). Western blot showed the effect of various concentrations of WHI-P154 on the phosphorylation of STAT3, AKT and ERK1/2 (f). Representative results are shown and similar results were obtained in two other independent trials.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4317847&req=5

fig03: The effect of WHI-P154 on the expression levels and localization of ABCG2, and on the blockade of STAT3, AKT and ERK1/2 phosphorylation in ABCG2-overexpressing cells. Western blot showed the effect of WHI-P154 on the expression level of ABCG2 after cells were treated with 4 μM of WHI-P154 for 0, 24, 48, and 72 h (a). Grayscale ratios of BCRP/β-actin were analyzed with Image J. The differences were statistically not significant (P > 0.05) (c). Similarly, WHI-P154 did not significantly alter the expression level of ABCG2 in ABCG2-482-R2 cells (b, d). Figure 3(a, c) representative result is shown from at least three independent experiments. Immunofluorescence staining showed membranous cellular localization of ABCG2 in H460/MX20 cells after treatment with WHI-P154 for 72 h. ABCG2 staining is shown in green. Fluorescent DAPI (blue) counterstains the nuclei (e). Western blot showed the effect of various concentrations of WHI-P154 on the phosphorylation of STAT3, AKT and ERK1/2 (f). Representative results are shown and similar results were obtained in two other independent trials.
Mentions: The reversal effect of ABCG2-mediated MDR could be achieved by either inhibiting ABCG2 function or decreasing the expression levels of ABCG2 protein. Therefore, Western blot analysis was performed to further ascertain the effect of WHI-P154 on the expression levels of ABCG2. Our results showed the expression levels of ABCG2 protein (Fig. 3) were not significantly altered in H460/MX20 cells after treatment with 4 μM WHI-P154 for 0, 24, 48 and 72 h, and the data also showed low-level expression of ABCG2 in parent H460 cells (Fig. 3a, c). The grayscale ratios of ABCG2/β-actin were proportional to the ABCG2 protein levels (Fig. 3c). Similarly, WHI-P154 did not significantly alter the expression levels of ABCG2 in ABCG2-482-R2 cells (Fig. 3b, d). In addition, we performed immunofluorescence to determine the effect of WHI-P154 on the cellular localization of ABCG2. As shown in Figure 3(e), WHI-P154 did not alter the localization of ABCG2. There results indicated that neither the expression levels nor localization of ABCG2 was significantly altered by WHI-P154.

Bottom Line: We found that WHI-P154 significantly enhanced the sensitivity of ABCG2-overexpressing cells to its substrates.WHI-P154 moderately sensitized ABCB1-overexpressing KB-C2 cells to its substrates whereas showed no sensitizing effect on ABCC1-, ABCC2 or ABCC10-mediated drug resistance.Collectively, these findings highlighted WHI-P154 could significantly reverse ABCG2-mediated multidrug drug resistance by directly blocking the efflux function.

View Article: PubMed Central - PubMed

Affiliation: Sun Yat-Sen University Cancer Center, State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Guangzhou, China; Department of Pharmaceutical Sciences, College of Pharmacy and Health Sciences, St John's University, Queens, New York, USA.

Show MeSH
Related in: MedlinePlus