Chai Xue, Zhang Jian-Wu, Li Sheng-Hui, Cheng Qing-Shui, Qin Ming-Ming, Yang Chun-Yan, Gao Jia-Lin, Huang Hou-Bao
Department of Urology, The First Affiliated Hospital of Wannan Medical College, Wuhu, Anhui, China.
Department of Pharmacology, North Sichuan Medical College, Nanchong, Sichuan, China.
Indian J Pathol Microbiol. 2021 Apr-Jun;64(2):294-301. doi: 10.4103/IJPM.IJPM_462_19.
Xanthoceraside is a component obtained in the husks of Xanthoceras sorbifolia Bunge. Series of researches proved that xanthoceraside had functions of anti-inflammation and anti-tumor effects. However, the mechanisms of xanthoceraside against bladder cancer are unclear. Accordingly, we proposed to investigate xanthoceraside's impacts and potential mechanisms in cells of bladder cancer.
By using the CCK-8 assay, we measured the viability of cells. With the use of 4,6-diamidino-2-phenylindole (DAPI) staining, we examined nuclear fragmentation and chromatin condensation in the nuclei of apoptotic cells. By using flow cytometry, we measured cell apoptosis. By using Western blotting, we tested the expressions of Caspase-9, Caspase-8, Caspase-3, Bcl-xL, P53, and PI3K/Akt/Bcl-2/Bax.
The proliferation of cell lines of human bladder cancer T24 and 5637 was suppressed by xanthoceraside significantly in a time- and concentration-dependent way. When cell lines 5637 and T24 were incubated as the xanthoceraside dose increased, the rates of cell apoptosis were upregulated, which was dependent on dose. According to further analysis, xanthoceraside induced apoptosis by upregulating Bax and downregulating the expression of Bcl-xL and Bcl-2. However, xanthoceraside did not change the expression of Caspase-9, Caspase-8, and Caspase-3. Interestingly, xanthoceraside also downregulated the expression of p-PI3K and p-Akt, and upregulated P53.
Xanthoceraside induces cell apoptosis through downregulation of the PI3K/Akt/Bcl-2/Bax signaling pathway in cell lines of human bladder cancer.
文冠果苷是从文冠果壳中提取的一种成分。一系列研究证明文冠果苷具有抗炎和抗肿瘤作用。然而，文冠果苷抗膀胱癌的机制尚不清楚。因此，我们提出研究文冠果苷对膀胱癌细胞的影响及其潜在机制。
通过CCK-8法检测细胞活力。使用4,6-二脒基-2-苯基吲哚（DAPI）染色，检测凋亡细胞核中的核碎裂和染色质凝聚。通过流式细胞术检测细胞凋亡。通过蛋白质免疫印迹法检测Caspase-9、Caspase-8、Caspase-3、Bcl-xL、P53以及PI3K/Akt/Bcl-2/Bax的表达。
文冠果苷能显著抑制人膀胱癌T24和5637细胞系的增殖，且呈时间和浓度依赖性。随着文冠果苷剂量增加，5637和T24细胞系的细胞凋亡率上调，且呈剂量依赖性。进一步分析表明，文冠果苷通过上调Bax并下调Bcl-xL和Bcl-2的表达诱导细胞凋亡。然而，文冠果苷并未改变Caspase-9、Caspase-8和Caspase-3的表达。有趣的是，文冠果苷还下调了p-PI3K和p-Akt的表达，并上调了P53的表达。
文冠果苷通过下调人膀胱癌细胞系中的PI3K/Akt/Bcl-2/Bax信号通路诱导细胞凋亡。
