CAJ | 학술논문

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。
목적：연구지방산합매（FAS）억제제천람균소（Cer）대이선암BXPC-3세포체외생장화조망적영향，병초보탐토가능적작용궤제。방법채용불동농도（0、2.5、5、10、20、40μg/ml）Cer 처리BXPC-3，세포증식여독성검측법（CCK8）검측OD치，계산대조조화각실험조적억제솔，병계산Cer IC50；취Cer농도위0、5、10、20μg/ml처리BXPC-3세포후진행류식세포술급AnnexinⅤ/PI조기조망검측세포조망솔화세포주기적변화；반전록취합매련반응（RT-PCR）검측FAS mRNA、Bcl-2 mRNA、Bax mRNA표체수평적변화；면역인적법（Western blot）검측세포조망인자Bcl-2、Bax단백표체정황。결과（1）Cer가명현억제BXPC-3세포적생장，병수착농도적증가화작용시간적연장이명현증강（P＜0.05）。（2）BXPC-3세포경Cer작용48 h후，세포주기S기비례상승，G2기세포비례하강（P＜0.05）。세포유명현조망，이조기조망위주。（3）BXPC-3세포경Cer작용48 h후，FAS mRNA、Bcl-2 mRNA적표체정농도의뢰성하강，Bax mRNA정농도의뢰성상승，각농도조여대조조비교차이유통계학의의（P＜0.05）。（4）Cer작용우BXPC-3세포48 h가사세포중적Bcl-2단백표체하강，Bax단백표체승고，차이구유통계학의의（P＜0.05）。결론 Cer 이시간、농도의뢰방식억제이선암 BXPC-3세포적생장，기궤제가능여조공세포조망상관기인Bcl-2、Bax적표체、조체세포주기、유도세포조망유관。
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(0,5,10,20μg/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 after 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-regulated 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.