中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2012年
5期
809-811
,共3页
卢振铎%刘真真%黄涛%焦得闯%乔江华%宋珂%崔树德
盧振鐸%劉真真%黃濤%焦得闖%喬江華%宋珂%崔樹德
로진탁%류진진%황도%초득틈%교강화%송가%최수덕
3′-脱氧腺苷%乳腺癌%脱噬作用%p53基因
3′-脫氧腺苷%乳腺癌%脫噬作用%p53基因
3′-탈양선감%유선암%탈서작용%p53기인
Cordycepin%Breast cancer%Apoptosis%p53 gene
目的 观察3′-脱氧腺苷对MDA-MB-231细胞生长、细胞周期、凋亡和p53表达的影响.方法 噻唑蓝(MTT)比色法观察3′-脱氧腺苷对体外培养MDA-MB-231细胞的生长抑制作用；流式细胞仪检测3′-脱氧腺苷作用于MDA-MB-231细胞48 h后细胞凋亡率和细胞周期变化；免疫组织化学方法检测MDA-MB-231细胞p53蛋白表达的变化.结果 MDA-MB-231细胞经3′-脱氧腺苷作用后,生长明显抑制,呈剂量和时间依赖性,以80 mg/L时作用最强；不同浓度(2、20、40、80、160mg/L)的3′-脱氧腺苷作用48h后,细胞凋亡率明显升高,其中在40 mg/L( 11.23±0.98)组、80 mg/L (15.21 ±2.0l)和160 mg/L( 13.27±2.26)与对照组(3.25±0.12)比较差异有统计学意义(P＜0.05).G0/G1期细胞比例逐渐升高,同时S期、G2/M期细胞比例相应减少(P＜0.05)；免疫组织化学染色结果表明,随着3′-脱氧腺苷浓度的增加,MDA-MB-231细胞p53蛋白表达水平逐渐降低.结论 3′-脱氧腺苷对MDA-MB-231细胞具有明显的生长抑制和促凋亡作用,这种作用可能是通过下调p53蛋白表达以及阻滞细胞周期而实现.
目的 觀察3′-脫氧腺苷對MDA-MB-231細胞生長、細胞週期、凋亡和p53錶達的影響.方法 噻唑藍(MTT)比色法觀察3′-脫氧腺苷對體外培養MDA-MB-231細胞的生長抑製作用；流式細胞儀檢測3′-脫氧腺苷作用于MDA-MB-231細胞48 h後細胞凋亡率和細胞週期變化；免疫組織化學方法檢測MDA-MB-231細胞p53蛋白錶達的變化.結果 MDA-MB-231細胞經3′-脫氧腺苷作用後,生長明顯抑製,呈劑量和時間依賴性,以80 mg/L時作用最彊；不同濃度(2、20、40、80、160mg/L)的3′-脫氧腺苷作用48h後,細胞凋亡率明顯升高,其中在40 mg/L( 11.23±0.98)組、80 mg/L (15.21 ±2.0l)和160 mg/L( 13.27±2.26)與對照組(3.25±0.12)比較差異有統計學意義(P＜0.05).G0/G1期細胞比例逐漸升高,同時S期、G2/M期細胞比例相應減少(P＜0.05)；免疫組織化學染色結果錶明,隨著3′-脫氧腺苷濃度的增加,MDA-MB-231細胞p53蛋白錶達水平逐漸降低.結論 3′-脫氧腺苷對MDA-MB-231細胞具有明顯的生長抑製和促凋亡作用,這種作用可能是通過下調p53蛋白錶達以及阻滯細胞週期而實現.
목적 관찰3′-탈양선감대MDA-MB-231세포생장、세포주기、조망화p53표체적영향.방법 새서람(MTT)비색법관찰3′-탈양선감대체외배양MDA-MB-231세포적생장억제작용；류식세포의검측3′-탈양선감작용우MDA-MB-231세포48 h후세포조망솔화세포주기변화；면역조직화학방법검측MDA-MB-231세포p53단백표체적변화.결과 MDA-MB-231세포경3′-탈양선감작용후,생장명현억제,정제량화시간의뢰성,이80 mg/L시작용최강；불동농도(2、20、40、80、160mg/L)적3′-탈양선감작용48h후,세포조망솔명현승고,기중재40 mg/L( 11.23±0.98)조、80 mg/L (15.21 ±2.0l)화160 mg/L( 13.27±2.26)여대조조(3.25±0.12)비교차이유통계학의의(P＜0.05).G0/G1기세포비례축점승고,동시S기、G2/M기세포비례상응감소(P＜0.05)；면역조직화학염색결과표명,수착3′-탈양선감농도적증가,MDA-MB-231세포p53단백표체수평축점강저.결론 3′-탈양선감대MDA-MB-231세포구유명현적생장억제화촉조망작용,저충작용가능시통과하조p53단백표체이급조체세포주기이실현.
Objective To investigate the effect of cordycepin on the expression of p53 gene and the growth,cell cycle and apoptosis of MDA-MB-231 cells.Methods The methyl thiazol tetrazolium (MTT) assay was applied to explore the impacts of cordycepin on the proliferation of MDA-MB-231 cells.After MDA-MB-231 cells were cultured with cordycepin (2,20,40,80,160 mg/L) for 48 h,the cell apoptosis rate and cell cycle were analyzed by flow cytometry.Immunocytochemistry was used to detect the protein expression of p53.Results Cordycepin could inhibit the growth of MDA-MB-231 cells in a dose-dependent manner,and there was the strongest effect in 80 mg/L concentration.Flow cytometry revealed cordycepin could increase the cell apoptosis rate,and there was significant difference between 40 mg/L (11.23 ±0.98),80 mg/L (15.21±2.01) and 160 mg/L (13.27 ±2.26) groups vs control group (3.25 ±0.12,P＜0.05),and the proportion of MDA-MB-231 cells in S phase and G2/M phase was gradually decreased but increased in G1 phase after culture with cordycepin (2,20,40,80,160 mg/L)for 48 h.Immunocytochemical results indicated that the p53 protein expression level in MDA-MB-231 cells was gradually reduced with the increase of the cordycepin concentration.Conclusion These results suggest that cordycepin can inhibit the cell growth and induce the cell apoptosis,which may be related to inhibition of the cell cycle and decrease in the expression of p53 in MDA-MB-231 cells.
