Tetrandrine induces apoptosis and blocks autophagic flux of bladder cancer T24 cells / 肿瘤

ABSTRACT

Objective: To investigate the effects of tetrandrine (Tet) on proliferation, apoptosis and autophagy of human bladder cancer cell lines, and explore its mechamism. Methods: Cell count kit-8 (CCK-8) assay was performed to analyze the effect of Tet on the proliferation of some bladder cancer cell lines and the half maximal inhibitory concentration (IC50) of Tet. After Tet treatment, the apoptosis rate of bladder cancer T24 cells was determined by flow cytometry, and the cleavage of apoptosis-related caspase-3 protein was detected by Western blotting. The activity of caspase was inhibited by Z-VAD-FMK, and then the viability of T24 cells treated by Tet was determined by CCK-8 assay. After the transfection of EGFP-LC3 into T24 cells, the cells were treated with Tet, and then the autophagosome formation was observed by confocal laser scanning microscopy assay. The expressions of autophagy-associated proteins LC3 and P62 in T24 cells after Tet treatment were detected by Western blotting. The effect of Tet on autophagic flux of T24 cells was determined by LC3 turnover assay, while the proliferative activity of T24 cells was detected by CCK-8 assay after treatment with 3-MA to inhibit autophagy. Results: Tet could obviously inhibit the proliferation of bladder cancer cells (P 50 of Tet (48 h) for cell lines 5637, T24, EJ and J82 were 8.03±1.2, 6.71 ±0.99, 4.93±0.72 and 2.72±0.24 μmol/L, respectively. Tet significantly induced apoptosis of T24 cells by caspase-3 activation (P 0.05). Conclusion: Tet can inhibit the proliferation of bladder cancer cells, induce the apoptosis, and block the autophagy.