Li Miao, Li Ping, Zhang Mei, Ma Feng, Su Li
Department of Chinese Medicine Tumor, the Affiliated Provincial Hospital of Anhui Medical University, Hefei 230001, China.
Zhongguo Fei Ai Za Zhi. 2014 Jun 20;17(6):444-50. doi: 10.3779/j.issn.1009-3419.2014.06.02.
It has been proven that Cyclin D1 and Cyclin E are the important positive regulators of cell cycle, they are closely related to the tumor proliferation. The aim of this study is to explore the relationship between Brucine and the proliferation in human lung cancer cell line PC-9, and the effect of it on the expression of Cyclin D1 and Cyclin E.
PC-9 cells were divided to 4 groups: the normal control group, the DMSO control group (2‰), the 150 μM Brucine group, and the 300 μM Brucine group. The proliferation rate of PC-9 cells was determined by The CellTiter-Glo Luminescent Cell Viability Assay and Colony Formation assay. The change of cell cycle was detected by Flow cytome try. Expressions of cell cycle regulators Cyclin D1, Cyclin E mRNA were determined by qRT-PCR. Protein expression of cell cycle regulators Cyclin D1, Cyclin E were determined by Western blot.
Compared with the control, Brucine remarkably inhibited the proliferation of PC-9 cells in a dose- and time-dependent manner (P<0.01); Flow cytome try showed that Brucine blocked the cell cycle of PC-9 cells at G0/G1, and the differences were statistically significant (P<0.01); qRT-PCR showed that the expression of Cyclin D1, Cyclin E mRNA were down-regulated; Western blot showed that the protein expression of Cyclin D1, Cyclin E were down-regulated.
Brucine can inhibit the proliferation of human lung cancer cell line PC-9 mainly by blocking the cell cycle at G0/G1 via down-regulating the expression of Cyclin D1, Cyclin E.
已证实细胞周期蛋白D1（Cyclin D1）和细胞周期蛋白E（Cyclin E）是细胞周期重要的正调控因子，它们与肿瘤增殖密切相关。本研究旨在探讨马钱子碱与人类肺癌细胞系PC-9增殖的关系，及其对Cyclin D1和Cyclin E表达的影响。
将PC-9细胞分为4组：正常对照组、二甲基亚砜（DMSO）对照组（2‰）、150μM马钱子碱组和300μM马钱子碱组。采用CellTiter-Glo发光细胞活力测定法和集落形成试验测定PC-9细胞的增殖率。通过流式细胞术检测细胞周期的变化。采用实时定量聚合酶链反应（qRT-PCR）测定细胞周期调节因子Cyclin D1、Cyclin E mRNA的表达。采用蛋白质印迹法测定细胞周期调节因子Cyclin D1、Cyclin E的蛋白表达。
与对照组相比，马钱子碱以剂量和时间依赖性方式显著抑制PC-9细胞的增殖（P<0.01）；流式细胞术显示马钱子碱将PC-9细胞的细胞周期阻滞在G0/G1期，差异具有统计学意义（P<0.01）；qRT-PCR显示Cyclin D1、Cyclin E mRNA的表达下调；蛋白质印迹法显示Cyclin D1、Cyclin E的蛋白表达下调。
马钱子碱可通过下调Cyclin D1、Cyclin E的表达，将细胞周期阻滞在G0/G1期，从而抑制人类肺癌细胞系PC-9的增殖。
