Department of Urology, Tongren Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200050, China.
J BUON. 2020 Jan-Feb;25(1):274-279.
Heptaphylline has been shown to suppress the growth of different types of cancer cells. Nonetheless, the anticancer effects of Heptaphylline have not been examined against human bladder cancer cells. Against this backdrop, this study was undertaken to investigate the anticancer effects of the carbazole alkaloid Heptaphylline against human bladder cancer cells.
Proliferation rate was determined by MTT assay. Apoptosis was demonstrated by DAPI and annexin V/propidium iodide (PI) assay. Electron microscopy was used for autophagy detection. Western blot analysis was used to determine protein expression.
The results showed that Heptaphylline suppressed the proliferation of the RT4 bladder cancer cells and exhibited an IC50 of 25 µM. The toxic effects of Heptaphylline were comparatively lower on the normal Hs172.T cells, as evidenced from the IC50 of 95 µM. The wound healing assay showed that Heptaphylline suppressed the migration of the RT4 bladder cancer cells. The DAPI and annexin V/PI staining showed that Heptaphylline induced apoptosis in the RT4 bladder cancer cells which was also accompanied by enhancement in the cleavage of PARP, caspase-3 and caspase-9. Additionally, Heptaphylline caused increase in Bax and decrease in Bcl-2 expression. Electron microscopic analysis showed that Heptaphylline also caused autophagy in the RT4 cells which was associated with increase in LC3, Atg5, Atg7 and Beclin-1 expression and decrease in p62 expression. This molecule also blocked the β-catenin signalling pathway in the RT4 bladder cancer cells.
Taken together, Heptaphylline suppressed the proliferation of the bladder cancer cells and may prove beneficial in the bladder cancer treatment.
水黄皮碱已被证明能抑制多种类型癌细胞的生长。然而，水黄皮碱对人膀胱癌细胞的抗癌作用尚未得到检验。有鉴于此，本研究旨在探讨咔唑生物碱水黄皮碱对人膀胱癌细胞的抗癌作用。
通过 MTT 测定法测定细胞增殖率。通过 DAPI 和 Annexin V/PI（PI）检测法证实细胞凋亡。使用电子显微镜检测自噬。采用 Western blot 分析测定蛋白表达。
结果表明，水黄皮碱抑制 RT4 膀胱癌细胞的增殖，其 IC50 为 25 μM。与 IC50 为 95 μM 的正常 Hs172.T 细胞相比，水黄皮碱对正常细胞 Hs172.T 的毒性作用相对较低。划痕愈合试验表明，水黄皮碱抑制 RT4 膀胱癌细胞的迁移。DAPI 和 Annexin V/PI 染色显示，水黄皮碱诱导 RT4 膀胱癌细胞凋亡，同时 PARP、caspase-3 和 caspase-9 的裂解增强。此外，水黄皮碱导致 Bax 表达增加，Bcl-2 表达减少。电镜分析显示，水黄皮碱还可引起 RT4 细胞自噬，与 LC3、Atg5、Atg7 和 Beclin-1 表达增加以及 p62 表达减少有关。该分子还阻断了 RT4 膀胱癌细胞中的β-连环蛋白信号通路。
综上所述，水黄皮碱抑制膀胱癌细胞的增殖，可能对膀胱癌的治疗有益。
