浅蓝菌素对人胰腺癌细胞株BXPC-3的影响及机制  

Effect and mechanism of cerulenin on human pancreatic carcinoma cell line BXPC-3

作　　者：许乐乐[1] 许春芳[2] 

机构地区：[1]苏州市立医院本部重症医学科,江苏省215006 [2]苏州大学附属第一医院消化内科

出　　处：《中华临床医师杂志（电子版）》2013年第19期86-89,共4页Chinese Journal of Clinicians(Electronic Edition)

基　　金：苏州市社会发展项目(SYS201115)

摘　　要：目的研究脂肪酸合酶(FAS)抑制剂浅蓝菌素(Cer)对胰腺癌BXPC-3细胞体外生长和凋亡的影响,并初步探讨可能的作用机制。方法采用不同浓度(0、2.5、5、10、20、40μg/ml)Cer处理BXPC-3,细胞增殖与毒性检测法(CCK8)检测OD值,计算对照组和各实验组的抑制率,并计算Cer IC50;取Cer浓度为0、5、10、20μg/ml处理BXPC-3细胞后进行流式细胞术及AnnexinⅤ/PI早期凋亡检测细胞凋亡率和细胞周期的变化;反转录聚合酶链反应(RT-PCR)检测FAS mRNA、Bcl-2 mRNA、Bax mRNA表达水平的变化;免疫印迹法(Western blot)检测细胞凋亡因子Bcl-2、Bax蛋白表达情况。结果 (1)Cer可明显抑制BXPC-3细胞的生长,并随着浓度的增加和作用时间的延长而明显增强(P<0.05)。(2)BXPC-3细胞经Cer作用48 h后,细胞周期S期比例上升,G2期细胞比例下降(P<0.05)。细胞有明显凋亡,以早期凋亡为主。(3)BXPC-3细胞经Cer作用48 h后,FAS mRNA、Bcl-2 mRNA的表达呈浓度依赖性下降,Bax mRNA呈浓度依赖性上升,各浓度组与对照组比较差异有统计学意义(P<0.05)。(4)Cer作用于BXPC-3细胞48 h可使细胞中的Bcl-2蛋白表达下降,Bax蛋白表达升高,差异具有统计学意义(P<0.05)。结论 Cer以时间、浓度依赖方式抑制胰腺癌BXPC-3细胞的生长,其机制可能与调控细胞凋亡相关基因Bcl-2、Bax的表达、阻滞细胞周期、诱导细胞凋亡有关。Objective To study the effect of Cerulenin on the growth and apoptosis of human pancreatic cancer cell line BXPC-3 in vitro and explore its possible mechanisms. Methods The growth of pancreatic cancer cell BXPC-3 treated by different concentrations of Cerulenin（0,2.5,5,10,20,40μg/ml） was evaluated by CCK8 assay and inhibition ratios were calculated in all groups and relative IC50 of Cerulenin were worked out by calculation; Using different concentrations of Cerulenin（0,5,10,20 μg/ml） to treat BXPC-3, the apoptosis ratioes and cell cycle shift were detected by flow cytometry and AnnexinV/PI; and the expression of FAS mRNA,Bcl-2 mRNA and Bax mRNA were detected by RT-PCR;Bcl-2,Bax proteins were detected by Western blot in vitro. Results （1） Cerulenin could significantly inhibit the growth of the human pancreatic cancer cell line BXPC-3, and the inhibitory effect had dose- and time- dependence （P〈0.05）. （2）The early stage of apoptosis and cyclomorphosis of BXPC-3 cells was detected atter they were incubated with cerulenin for 48 hours. With the increase of Cer concentration, the ratio of S phase increased and ratio of G2 phase declined homologously（P〈0.05）, Appearantly, the apoptosis rate was positively correlated to Cer concentration.（3）The expression of FAS mRNA and Bcl-2 mRNA were decreased with concentration-dependence after BXPC-3 cells were incubated 48 hours with Cer, but the expression of Bax mRNA was increased. Comparisons of all Cer groups and control group have significant difference（P〈 0.05）.（4）The protein level of Bcl-2 was down-regulated and that of Bax was up-regnlated significantly after BXPC-3 cells were incubated with Cerfor 48 hours（P〈0.05）. Conclusion Cerulenin could inhibit the growth of human pancreatic cancer cell line BXPC-3 in a dose-and-time dependent manner, and its mechanisms may directly inhibit tumor cells to proliferate and arrest cell cycle in S stage, and induce early apoptosis.

关 键 词：胰腺肿瘤 变蓝菌素 BCL-2相关X蛋白质 Bcl-2 

分 类 号：R735.9[医药卫生—肿瘤]