Huang Yi-Qiang, Zhang Yi-Ming, Jin Zhong, Li Xie-Zhao, Wang Chong-Shan, Xu Kai, Huang Peng, Liu Chun-Xiao
Department of Urology, Zhujiang Hospital, Southern Medical University, Guangzhou 510280, China. E-mail:
Nan Fang Yi Ke Da Xue Xue Bao. 2016 Jun;36(6):857-62.
To investigate the antitumor effect of lycorine on renal cell carcinoma ACHN cells and explore the possible mechanism.
We used flow cytometry to examine the effect of lycorine on ACHN cell cycle and apoptosis. The cell proliferation, migration and invasion were assessed with MTS assay, wound healing assay, and Transwell assay, respectively. Colony forming assay was performed, and the mRNA and protein levels of Bax, Bcl-2, survivin, caspase-3, cyclin D1 and CDK4 were measured with qRT-PCR and Western blotting.
Lycorine obviously inhibited the proliferation of ACHN cells with an IC(50) of 24.34 µmol/L. Lycorine also induced apoptosis of ACHN cells, caused cell cycle arrest at G(0)/G(1) phase, and suppressed the colony forming ability of the cells in a dose-dependent manner. The migration and invasion of ACHN cells were significantly inhibited by 5 µmol/L lycorine. Lycorine up-regulated the mRNA levels of CDK4, Bax, caspase-3 while down-regulated the levels of survivin, Bcl-2 and Cyclin D1; the protein levels of CDK4 and Bax were increased and cyclin D1, Bcl-2 and surviving expressions were decreased, but caspase-3 expression showed no significant changes following the treatment.
Lycorine has obvious antitumor effect against ACHN cells, suggesting its value as a new therapeutic agent for renal cell carcinoma.
研究石蒜碱对肾细胞癌ACHN细胞的抗肿瘤作用，并探讨其可能的作用机制。
我们采用流式细胞术检测石蒜碱对ACHN细胞周期和凋亡的影响。分别用MTS法、伤口愈合试验和Transwell试验评估细胞增殖、迁移和侵袭能力。进行集落形成试验，并用qRT-PCR和蛋白质印迹法检测Bax、Bcl-2、survivin、caspase-3、细胞周期蛋白D1和细胞周期蛋白依赖性激酶4（CDK4）的mRNA和蛋白质水平。
石蒜碱明显抑制ACHN细胞的增殖，IC50为24.34 μmol/L。石蒜碱还诱导ACHN细胞凋亡，使细胞周期停滞于G0/G1期，并以剂量依赖性方式抑制细胞的集落形成能力。5 μmol/L石蒜碱显著抑制ACHN细胞的迁移和侵袭。石蒜碱上调CDK4、Bax、caspase-3的mRNA水平，同时下调survivin、Bcl-2和细胞周期蛋白D1的水平；处理后CDK4和Bax的蛋白质水平升高，细胞周期蛋白D1、Bcl-2和survivin的表达降低，但caspase-3的表达无明显变化。
石蒜碱对ACHN细胞具有明显的抗肿瘤作用，提示其作为肾细胞癌新型治疗药物的价值。
