Effects of tea polyphenols combined with epirubicin on T24 bladder cancer cell line and its mechanism
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Effects of tea polyphenols combined with epirubicin on T24 bladder cancer cell line and its mechanism
Journal of Chongqing Medical UniversityVol. 40, Issue 8, Pages: 1078-1083(2015)
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重庆医科大学附属第一医院泌尿外科,重庆,400016
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Published:2015,
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Yang Fan, He Weiyang, Zhu Xin, et al. Effects of tea polyphenols combined with epirubicin on T24 bladder cancer cell line and its mechanism. [J]. Journal of Chongqing Medical University 40(8):1078-1083(2015)
DOI:
Yang Fan, He Weiyang, Zhu Xin, et al. Effects of tea polyphenols combined with epirubicin on T24 bladder cancer cell line and its mechanism. [J]. Journal of Chongqing Medical University 40(8):1078-1083(2015)DOI:
Effects of tea polyphenols combined with epirubicin on T24 bladder cancer cell line and its mechanism
Objective:To investigate the effect of tea polyphenols(TP) on epirubicin(EPI)-induced cell proliferation and cell apopto-sis of human bladder cancer cells T24 in vitro and its mechanism. Methods:MTT assay was applied to investigate the effects of differ-ent levels of tea polyphenols or epirubicin on the T24 cells. The experiment was divided into 4 groups:control group(PBS),TP group(88.8 μmol/L),EPI group(4.6 μmol/L),and TP(88.8 μmol/L) plus EPI(4.6 μmol/L) group. The proliferation inhibition rate was as-sayed by MTT;Annexin V-FITC/PI double staining flow cytometry was used to analyze apoptosis. The ultra-structural cellular changes were observed by transmission electron microscope(TEM);The formation of MAP1LC3-Ⅱ puncta was examined after transfection of a GFP-LC3 plasmid. The levels of LC-3-Ⅱ,P62,CASP3,PARP proteins were detected by Western blot,respectively. Results:Treatment with TP(88.8 μmol/L) and EPI(4.6 μmol/L) leaded to a significant increase of inhibiting rate(P=0.000) and a dramatic improvement in apoptotic changes compared to single EPI or TP group(P=0.000). Autophagy was observed obviously in bladder cancer cells T24 treated with EPI after 8 h. TEM examination revealed the appear-ance of autophagosome with double-membrane structure. Im-munofluorescence showed that EPI strongly augmented the number of cells with increased GFP-LC3 puncta and GFP-LC3 puncta formation within the cell. Western blot showed the ex-pression of LC-3-Ⅱ was increased in T24 cells after treated with EPI,while the expression of P62 was decreased. TP com-bined with EPI and single groups both induced the expression of active apoptosis proteins,and the effect was more obvious in TP plus EPI group. Conclusion:TP combined with EPI can synergisti-cally inhibit the proliferation and promote cell apoptosis,which may be associated with the reduction of EPI-induced autophagy by TP.
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