摘要
目的观察表皮生长因子受体（EGFR）抑制剂埃罗替尼对体外培养的胰腺癌细胞BxPC3生长的影响，并探讨其作用机制。方法应用MTT法检测埃罗替尼作用后BxPC3细胞的增殖情况；用流式细胞分析、透射电镜和原位末端标记（TUNEL）法观察细胞凋亡和细胞周期的变化；RT—PCR法检测细胞bcl-2、bax、bcl-xl、bak mRNA表达。结果埃罗替尼呈剂量和时间依赖性抑制BxPC3细胞生长。72h后，1、100μmol／L的埃罗替尼处理组细胞存活率分别为（90．25±2．62）％和（40．75±2．98）％。两者比较具有统计学意义（P〈0．01）。50μmol／L埃罗替尼处理BxPC3后24、96h的细胞存活率分别为（74．0±4．08）％和（49．50±1．29）％，两者比较也具有统计学意义（P〈0．01）。50μmol／L埃罗替尼处理组的细胞凋亡率为（11．0±1．1）％，显著高于对照组的（6．2±1．1）％（P〈0．01）；G0／G1细胞占（73．4±1．3）％，也显著高于对照组的（63．3±1．0）％；透射电镜可见细胞呈现明显凋亡形态，并见凋亡小体形成。埃罗替尼处理组细胞bcl-2、bcl—xl mRNA表达下调，bax mRNA表达轻微上调，bak mRNA的表达不受影响。结论EGFR抑制剂埃罗替尼体外可抑制胰腺癌细胞系BxPC3的生长，其机制可能与阻滞细胞周期，上调促凋亡蛋白和下调凋亡抑制蛋白有关.
Objective To investigate the effect and possible mechanism of erlotinib, an epidermal growth factor receptor inhibitor, on human pancreatic cancer cell lines BxPC3 in vitro. Methods Methylthiazolyltetrazolium （MTY） assay was used to detected the proliferation of BxPC3 after exposure to erlotinib, apoptosis and cell cycle changes were studied by flow cytometry and terminal deoxynucleotidyl transferase-mediated nick end labeling assay （TUNEL）. The expressions of bel-2 mRNA, bax mRNA, bcl-xL mRNA and bak mRNA were determined by reverse transcriptase polymerase chain reaction （RT-PCR）. Results Erlotinib inhibited BxPC3 cells growth in a dose and time dependent manner in vitro. The cell viabilities in erlotinib 1 μmol/L and 100 μ moL/L groups 72 h later were （90.25 ± 2.62 ）% and （40.75 ± 2.98 ） % , and the difference was statistically significant （ P 〈 0.01 ）. The cell viability in erlotinib 50 μmol/L groups 24 h and 96 h after BxPC3 exposure were （74.0 ± 4.08 ） % and （49.50 ± 1.29 ） % , and the difference was statistically significant （P 〈0.01 ）. Cell apoptosis rate in erlotinib 50 μmol/L group was（ 11.0 ± 1.1 ）%, which was significantly higher than （6.2 ± 1. 1 ） % in control group （P 〈 0.01 ）. G0/G1 cell accounted for （73.4 ± 1.3） % of all the cells, which was significantly higher than （63.3 ± 1.0） % in control group. With transmission eleetron microscope, the morphology of BxPC3 cells showed typical apoptosis and apoptotic body. The expressions of bcl-2 mRNA, bel-xl mRNA were down-regulated, while the expression of bax mRNA was slightly up-regulated, and the expression of bak mRNA was not affected. Conclusions The growth of BxPC3cells could be suppressed by erlotinib and possible mechanisms involved blocking cell cycle, up-regulating apoptosis proteins and down-regulating apoptosis inhibitor proteins.
出处
《中华胰腺病杂志》
CAS
2009年第6期395-398,共4页
Chinese Journal of Pancreatology
关键词
胰腺肿瘤
受体
表皮生长因子
细胞凋亡
埃罗替尼
Pancreatic neoplasms
Receptor, epidermal growth factor
Apoptosis
Erlotinib
