Differential expression of P-gp in bladder cancer cell lines
10.3760/cma.j.issn.1009-9158.2015.04.015
- VernacularTitle:P-gp在膀胱癌细胞中的差异表达研究
- Author:
Man ZHAO
;
Man ZHANG
;
Shuliang YU
;
Min ZHANG
;
Ping MA
- Publication Type:Journal Article
- Keywords:
Urinary bladder neoplasms;
Carcinoma,transitional cell;
Cell line,tumor;
R Glycoprotein
- From:
Chinese Journal of Laboratory Medicine
2015;38(4):277-280
- CountryChina
- Language:Chinese
-
Abstract:
Objective The generation of drug resistance often leads to the failure of the bladder cancer chemotherapy.P-glycoprotein (P-gp) is an ATP-dependent drug efflux pump linked to development of multidrug resistance in cancer cells.The laboratory has successfully established adriamycin-resistant human bladder cancer cell line (pumc-91/ADM) from its parental cell line (pumc-91).According to the drug resistant spectrum analysis,pumc-91/ADM cell line exhibited the characteristics of multi-drug resistance.However,the expression of P-gp in two cell lines was still unknown.In this paper,there was a comparison between pumc-91/ADM and pumc-91 about the differential expression of P-gp.Methods To determine the expression and location of P-gp in pumc-91 and pumc-91/ADM,qRT-PCR,Western blot and immunocytochemistry were applied in the experiment.qRT-PCR was implemented to research the expression of P-gp mRNA in two cell lines (pumc-91/ADM and pumc-91).Western blot was adopted to investigate the expression of P-gp protein in pumc-91 and pumc-91/ADM cell lines.Immunocytochemistry technique was used to explore the cellular location of P-gp and affirm its expression in two cell lines visually.Student's t-test was employed for statistical analysis and P ＜ 0.05 was considered statistically significant.Results qRT-PCR analysis revealed that the expression of P-gp mRNA was upregulated in drug-resistant cell line pumc-91/ADM compared to parental cell line pumc-91.To normalize for differences in the amount of total RNA,GAPDH was selected as an endogenous RNA control.Compared with pumc-91,the expression of P-gp mRNA was upregulated 7.74 fold in pumc-91/ADM (t =11.97,P ＜ 0.05).Consistent with the qRT-PCR result,Western blot confirmed the protein of P-gp expressed differentially in two cell lines.The expression of P-gp protein was significantly increased in pumc-91/ADM compared to pumc-91.According to the results,the differences between pumc-91 and pumc-91/ADM had statistical significance (t =4.35,P＜0.05).Immunocytochemical analysis results demonstrated that P-gp was not only located in cell membrane but also in cytoplasm of the two cell lines.The expression of P-gp in pumc-91/ADM increased distinctly.The difference was statistically significant (t =11.41,P ＜ 0.05).Conclusion Compared with pumc-91,the expression of P-gp in pumc-91/ADM was significantly upregulated.
