二十二碳六烯酸对人胆囊癌GBC-SD细胞系生长的抑制作用

目的探讨二十二碳六烯酸（DHA）对体外培养的人胆囊癌GBC-SD细胞增殖的影响。方法采用体外细胞培养的方法,在处于指数生长期的GBC-SD细胞培养剂RPMI 1640中分别添加12.5μg/ml、25μg/ml、50μg/ml的DHA,培养24～72小时后,采用噻唑蓝法（MTT法）检测GBC-SD细胞的增殖情况,光镜下观察细胞形态变化。采用DNA琼脂糖凝胶电泳、Annexin V-FITC/PI染色荧光显微镜检测GBC-SD细胞的凋亡情况。结果经12.5μg/ml的DHA作用24～72小时后GBC-SD细胞的A值与对照组比较差别无统计学意义（P〉0.05）,光镜下细胞形态无明显改变.经25μg/ml、50μg/ml的DHA作用24～72小时后GBC-SD细胞的A值与对照组比较差别有统计学意义（P〈0.05）,光镜下见细胞皱缩、核染色质凝聚、边集。细胞经25μg/ml、50μg/ml的DHA作用24小时后,DNA琼脂糖凝胶电泳显示DNA裂解片段出现阶梯状条带,经Annexin V-FITC/PI染色荧光显微镜下观察与对照组比较见较多绿色荧光。结论DHA可以抑制人胆囊癌GBC-SD细胞系的增殖并诱导其凋亡。

The inhibitory effect of docosahexaenic acid on the growth of human gallbladder carcinoma GBC-SD cell line

Objective To investigate the effect of docosahexaenic acid（DHA） on the growth of human gallbladder carcinoma GBC-- SD cell. Methods By cells culturing in vitro, the human gallbladder carcinoma GBC--SD cells of logarithmic growth phase was cultured in RPMI1640 medium supplemented with DHA at concentration 12. 5ug/ml, 25 ug/ml. 50ug/ml respectively. After incubation from 24h to 72h,cell viability and proliferation was determined by MTT colourimetric assay, the cellular morphology was observed through microscope. Agarose gel electrophoresis and stained by Annexin V--FITC/PI fluorescence microscopy were used to detect apoptosis. Results After incubation with 12.5ug/ml DHA for 24h,48h and 72h, the absorbance of GBC--SD cells was not affected,the cellular morphology was not changed significantly through microscope. After incubation with 25 ug/ml. 50ug/ml DHA for 24h, 48h and 72h, the absorbance of GBC--SD cells was inhibited significantly, cell shrinkage and chromatin con- densation could be seen through microscope. After incubation with25 ug/ml.50 ug/ml DHA for 24h,a DNA ladder was appeared when ran on an agarose gel. After incubation with25ug/ml.50 ug/ml DHA for 24h, GBC--SD cell stained by Annexin V--FITC/PI,more green fluorescence could be seen through fluorescence microscopy. Conclusions DHA can inhibit the growth of human gallbladder carcinoma GBC- SD cell line and induce the apoptosis.