CAJ | 학술논문

目的：探讨二十碳五烯酸（EPA）诱导人结肠癌SW480细胞凋亡的作用机制。方法体外培养SW480细胞，不同浓度 EPA 作用于 SW480细胞后，采用流式细胞仪检测 SW480细胞线粒体膜电位的变化，利用试剂盒检测细胞色素 C 含量变化，Western blot 法检测活化的 caspase-9和 caspase-3蛋白表达水平。结果经不同剂量 EPA（0μg／mL、42．1μg／mL、84．2μg／mL、168．4μg／mL）处理后，SW480细胞增殖受到明显抑制， EPA 呈剂量依赖性的诱导 SW480细胞线粒体膜电位下降，分别为（99．71±0．04）％、（95．04±0．10）％、（88．65±0．41）％、（73．60±1．20）％，与对照组相比，差异均有统计学意义（t ＝5．161、6．302、4．601、5．198，均P ＜0．05）。细胞浆中细胞色素 C 含量分别为（12．8±1．2）ng／mL、（115．5±3．5）ng／mL、（290．5±5．2）ng／mL、（262．0±12．5）ng／mL），均明显高于对照组，差异均有统计学意义（t ＝6．345、6．013、5．846、4．613，均 P ＜0．01）；随着 EPA 浓度的增加，活化 caspase-9和 caspase-3蛋白水平呈上调趋势。结论EPA 体外能够诱导SW480细胞发生凋亡，其作用机制主要通过线粒体介导的内源性凋亡途径，细胞色素 C 的释放，caspase-9和caspase-3蛋白活化。
목적：탐토이십탄오희산（EPA）유도인결장암SW480세포조망적작용궤제。방법체외배양SW480세포，불동농도 EPA 작용우 SW480세포후，채용류식세포의검측 SW480세포선립체막전위적변화，이용시제합검측세포색소 C 함량변화，Western blot 법검측활화적 caspase-9화 caspase-3단백표체수평。결과경불동제량 EPA（0μg／mL、42．1μg／mL、84．2μg／mL、168．4μg／mL）처리후，SW480세포증식수도명현억제， EPA 정제량의뢰성적유도 SW480세포선립체막전위하강，분별위（99．71±0．04）％、（95．04±0．10）％、（88．65±0．41）％、（73．60±1．20）％，여대조조상비，차이균유통계학의의（t ＝5．161、6．302、4．601、5．198，균P ＜0．05）。세포장중세포색소 C 함량분별위（12．8±1．2）ng／mL、（115．5±3．5）ng／mL、（290．5±5．2）ng／mL、（262．0±12．5）ng／mL），균명현고우대조조，차이균유통계학의의（t ＝6．345、6．013、5．846、4．613，균 P ＜0．01）；수착 EPA 농도적증가，활화 caspase-9화 caspase-3단백수평정상조추세。결론EPA 체외능구유도SW480세포발생조망，기작용궤제주요통과선립체개도적내원성조망도경，세포색소 C 적석방，caspase-9화caspase-3단백활화。
Objective To investigate the effect of eicosapentaenoic acid(EPA)on the apoptosis of human colon cancer SW480 cells and the mechanisms.Methods Mitochondrial membrane potential was detected,the quan-tity of cytochrome C was analyzed by Elisa kit,and the expression of cleaved caspase -9 and caspase -3 was detected by Western Blot.Results After treatment with EPA (0μg/mL,42.1μg/mL,84.2μg/mL,168.4μg/mL),the mitochondrial membrane potential(Δψm)of SW480 cells were declined (P <0.05 ),the values were (99.71 ± 0.04)%,(95.04 ±0.10)%,(88.65 ±0.41)% and (73.60 ±1.20)%(t =5.161,6.302,4.601,5.198,all P <0.05).The quantity of cytochrome C in cytosol was increased significantly compared with no treatment group,and the values were (12.8 ±1.2)ng/mL,(115.5 ±3.5)ng/mL,(290.5 ±5.2)ng/mL and (262.0 ±12.5 )ng/mL in different EPA treatment groups(t =6.345,6.013,5.846,4.613,all P <0.01).The expression of cleaved caspase -9 and caspase -3 were significantly increased.Conclusion EPA inhibits SW480 cells growth and induces apoptosis in a dose dependent manner.This action may be mediated by mitochondria -mediated intrinsic apoptosis pathway,cyto-chrome C release,and caspase -9 and caspase -3 activation.