ABSTRACT The inhibitory effect of buforin IIb on different types of cancer, although not liver cancer, has been demonstrated previously. The aim of the present study was to investigate the effects of buforin IIb on the progression of liver cancer. The human liver cancer cell line HepG2 was treated with purified buforin IIb and the cell activity was determined by MTT, colony formation and transwell assays. The protein expression levels of cyclin-dependent kinases (CDKs) and cyclins were analyzed by western blotting and immunofluorescent cell staining. A tumor growth model was constructed using nude mice, and buforin IIb treatment was administered. The levels of CDK2 and cyclin A in the tumor tissues were detected by western blotting. Buforin IIb treatment depressed cell viability and colony formation and induced apoptosis significantly, and 1.0 µM concentration of buforin IIb was found to be the optimal dosage. The cell cycle was arrested at the G2/M phase following buforin IIb treatment. CDK2 and cyclin A were downregulated by treatment of the cells with 1.0 µM buforin IIb for 24 h. Treatment with buforin IIb also inhibited the migration of liver cancer cells in vitro. Furthermore, 50 nmol buforin IIb injection suppressed HepG2 cell subcutaneous tumor growth in the nude mouse model. Similar to the in vitro results, buforin IIb injection reduced the expression of CDK2 and cyclin A in the tumor tissue. these results demonstrate that buforin IIb inhibited liver cancer cell growth via the regulation of CDK2 and cyclin A expression.

中文翻译：

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Buforin IIb 诱导肝癌细胞周期停滞

摘要 buforin IIb 对不同类型癌症（尽管不是肝癌）的抑制作用先前已得到证实。本研究的目的是调查buforin IIb对肝癌进展的影响。人肝癌细胞系 HepG2 用纯化的 buforin IIb 处理，细胞活性通过 MTT、集落形成和 transwell 测定确定。通过蛋白质印迹和免疫荧光细胞染色分析细胞周期蛋白依赖性激酶 (CDK) 和细胞周期蛋白的蛋白质表达水平。使用裸鼠构建肿瘤生长模型，并给予buforin IIb治疗。通过蛋白质印迹检测肿瘤组织中CDK2和细胞周期蛋白A的水平。Buforin IIb 处理抑制细胞活力和集落形成并显着诱导细胞凋亡，并且 1. 发现 0 µM 浓度的 buforin IIb 是最佳剂量。buforin IIb 治疗后，细胞周期在 G2/M 期停滞。CDK2 和细胞周期蛋白 A 通过用 1.0 µM buforin IIb 处理细胞 24 小时而下调。用buforin IIb 治疗也抑制了体外肝癌细胞的迁移。此外，50 nmol buforin IIb 注射抑制了裸鼠模型中 HepG2 细胞皮下肿瘤的生长。与体外结果相似，buforin IIb注射降低了肿瘤组织中CDK2和cyclin A的表达。这些结果表明，buforin IIb 通过调节 CDK2 和细胞周期蛋白 A 的表达来抑制肝癌细胞的生长。CDK2 和细胞周期蛋白 A 通过用 1.0 µM buforin IIb 处理细胞 24 小时而下调。用buforin IIb 治疗也抑制了体外肝癌细胞的迁移。此外，50 nmol buforin IIb 注射抑制了裸鼠模型中 HepG2 细胞皮下肿瘤的生长。与体外结果相似，buforin IIb注射降低了肿瘤组织中CDK2和cyclin A的表达。这些结果表明，buforin IIb 通过调节 CDK2 和细胞周期蛋白 A 的表达来抑制肝癌细胞的生长。通过用 1.0 µM buforin IIb 处理细胞 24 小时，CDK2 和细胞周期蛋白 A 被下调。用buforin IIb 治疗也抑制了体外肝癌细胞的迁移。此外，50 nmol buforin IIb 注射抑制了裸鼠模型中 HepG2 细胞皮下肿瘤的生长。与体外结果相似，buforin IIb注射降低了肿瘤组织中CDK2和cyclin A的表达。这些结果表明，buforin IIb 通过调节 CDK2 和细胞周期蛋白 A 的表达来抑制肝癌细胞的生长。与体外结果相似，buforin IIb注射降低了肿瘤组织中CDK2和cyclin A的表达。这些结果表明，buforin IIb 通过调节 CDK2 和细胞周期蛋白 A 的表达来抑制肝癌细胞的生长。与体外结果相似，buforin IIb注射降低了肿瘤组织中CDK2和cyclin A的表达。这些结果表明，buforin IIb 通过调节 CDK2 和细胞周期蛋白 A 的表达来抑制肝癌细胞的生长。