摘要
目的探讨麦胚凝集素(WGA)对人乳腺癌MDA-MB-231细胞的生长及凋亡基因表达的影响。方法实验分为对照组、0.035μmol/LWGA组、0.07μmol/LWGA组、0.14μmol/LWGA组和0.28μmol/LWGA组,应用酸性磷酸酶法(APA)、流式细胞术(FCM)和Western blotting法,探讨WGA对MDA-MB-231细胞生长、细胞凋亡率、线粒体膜电位、细胞凋亡相关基因蛋白聚腺苷酸二磷酸核糖转移酶(PARP)、细胞色素C表达的影响。结果不同浓度WGA作用24、48和72h时可抑制MDA-MB-231细胞生长;与人乳腺上皮细胞HBL-100相比,WGA对MDA-MB-231细胞的抑制作用尤为显著。光镜下观察0.14μmol/LWGA组与0.28μmol/LWGA组作用24h时,细胞密度稀疏,体积缩小;早期凋亡率分别为(28.7±3.48)%和(30.15±3.96)%;晚期凋亡率分别为(53.66±4.21)%和(63.18±5.53)%;线粒体膜电位平均荧光强度显著降低;凋亡相关蛋白PARP呈现剪切带,细胞色素C表达上调。结论 WGA抑制人乳腺癌MDA-MB-231细胞增殖,通过干预线粒体途径诱导细胞发生凋亡。
Objective To study the influence of wheat germ agglutinin （WGA） on cell growth and expressions of apoptosis genes in MDA-MB-231 cells. Methods MDA-MB-231 cells were cultured in DMEM containing 10% fetal bovine serum at 37℃ in a humidified atmosphere with 5% CO2. The experiment was performed on five groups： WGA groups were given 0. 035, 0. 07,0.14,0. 28μmol/L WGA respectively, while in the control group the cells were merely grown in DMEM medium solution. Acid phosphatase assay （APA） , flow cytometry（FCM） and Western blotting were used to detect the changes of cell viability, apoptosis ratio, mitochondrial transmembrane potential, the expressions of apoptosisrelated protein poly ADP-ribose polymerase （PARP） and cytochrome C. Each experiment was done four times independently. Results Cell growth was inhibited after MDA-MB-231 cells were co-cultured with different concentrations of WGA for 24, 48, and 72 hours. Compared with human breast epithelia cells line HBL-100, MDA-MB-231 cells were more sensitive to WGA. After cells were treated with WGA for 24 hours, cell numbers reduced, volumes diminished. Early apoptosis ratio caused by WGA （0. 14μmo//L, 0. 28μmol/L, 24 hours） were （ 28.7 ± 3.48 ）% and （ 30. 15 ± 3.96）% , while the late apoptosis rates were （ 53.66 ± 4.21 ） % and （ 63.18 ± 5.53 ） %. Mean fluorescence intensity （MFI） of mitochondrial transmembrane potentials in MDA-MB-231 cells decreased. Apoptosis-related protein PARP showed an cleavage and cytochrome C expression increased in a dose-depandance manner. Conclusion WGA could significally restrain cell growth of breast cancer cell line MDA-MB-231. WGA can induce cleavage of PARP and activate mitochondrial signaling pathway which would induce apoptosis,but the mechanism of WGA is yet to be studied in detail.
出处
《解剖学报》
CAS
CSCD
北大核心
2010年第4期554-558,共5页
Acta Anatomica Sinica
基金
教育部教育振兴行动计划专项资助项目(985222016224)
