肿瘤药学
腫瘤藥學
종류약학
ANTI-TUMOR PHARMACY
2014年
5期
332-335
,共4页
F200%乳腺癌%凋亡%细胞周期蛋白依赖性激酶
F200%乳腺癌%凋亡%細胞週期蛋白依賴性激酶
F200%유선암%조망%세포주기단백의뢰성격매
F200%Breast cancer%Apoptosis%Cell cycle dependent kinase
目的：研究细胞周期依赖性激酶（cell cycle dependent kinase，CDK）CDK9抑制剂F200对乳腺癌细胞MCF7凋亡的影响。方法 MCF7细胞培养于含0.01 mg·mL-1人重组胰岛素及10％胎牛血清的RPMI 1640培养液中，待对数生长期时接种细胞进行实验，24 h贴壁后给药：分为阴性对照组（0.5％ DMSO）、阳性对照组（R-Roscovitine 5.66μM）及药物组（F2000.1μM，0.71μM），给药48 h后利用TUNEL法染色观察细胞凋亡DNA断裂情况及细胞凋亡形态学变化、流式细胞技术检测细胞的凋亡比率、免疫印迹法检测凋亡标志蛋白PARP的表达情况。结果 TUNEL结果显示，随着F200浓度增加，细胞出现明显的固缩变圆，细胞核可见深染致密颗粒，细胞核质分界明显等凋亡表现；流式细胞仪结果显示，0.71μM的F200能够诱导32.6％的细胞凋亡；Western Blot结果显示，随着F200浓度增加，PARP蛋白剪切水平明显增加。结论 F200能有效促进乳腺癌细胞MCF7的凋亡。
目的：研究細胞週期依賴性激酶（cell cycle dependent kinase，CDK）CDK9抑製劑F200對乳腺癌細胞MCF7凋亡的影響。方法 MCF7細胞培養于含0.01 mg·mL-1人重組胰島素及10％胎牛血清的RPMI 1640培養液中，待對數生長期時接種細胞進行實驗，24 h貼壁後給藥：分為陰性對照組（0.5％ DMSO）、暘性對照組（R-Roscovitine 5.66μM）及藥物組（F2000.1μM，0.71μM），給藥48 h後利用TUNEL法染色觀察細胞凋亡DNA斷裂情況及細胞凋亡形態學變化、流式細胞技術檢測細胞的凋亡比率、免疫印跡法檢測凋亡標誌蛋白PARP的錶達情況。結果 TUNEL結果顯示，隨著F200濃度增加，細胞齣現明顯的固縮變圓，細胞覈可見深染緻密顆粒，細胞覈質分界明顯等凋亡錶現；流式細胞儀結果顯示，0.71μM的F200能夠誘導32.6％的細胞凋亡；Western Blot結果顯示，隨著F200濃度增加，PARP蛋白剪切水平明顯增加。結論 F200能有效促進乳腺癌細胞MCF7的凋亡。
목적：연구세포주기의뢰성격매（cell cycle dependent kinase，CDK）CDK9억제제F200대유선암세포MCF7조망적영향。방법 MCF7세포배양우함0.01 mg·mL-1인중조이도소급10％태우혈청적RPMI 1640배양액중，대대수생장기시접충세포진행실험，24 h첩벽후급약：분위음성대조조（0.5％ DMSO）、양성대조조（R-Roscovitine 5.66μM）급약물조（F2000.1μM，0.71μM），급약48 h후이용TUNEL법염색관찰세포조망DNA단렬정황급세포조망형태학변화、류식세포기술검측세포적조망비솔、면역인적법검측조망표지단백PARP적표체정황。결과 TUNEL결과현시，수착F200농도증가，세포출현명현적고축변원，세포핵가견심염치밀과립，세포핵질분계명현등조망표현；류식세포의결과현시，0.71μM적F200능구유도32.6％적세포조망；Western Blot결과현시，수착F200농도증가，PARP단백전절수평명현증가。결론 F200능유효촉진유선암세포MCF7적조망。
Objective This study aimed to investigate the effects of cell cycle dependent kinase inhibitor F200 on ap-optosis of human breast cancer cell MCF7. Methods MCF7 cells were cultured in RPMI-1640 medium with 0.01 mg·mL-1 human recombinant insulin and 10%fetal bovine serum. Cells were subcultured at exponential growth phase. 24 h later, cells were adherent to the plate and treatments were given according to groups setup, negative control (0.5%DMSO), posi-tive control (R-Roscovitine 5.66μM), F200 groups (F200 0.1μM, 0.71μM). At 48 h after treatment, cell morphological change of apoptosis was measured by TUNEL methods and the apoptosis rate was detected by flow cytometry. The expres-sion of PRAP was measured by Western Bolt. Results The TUNEL results showed that as the F200 concentration increased, the cell apoptosis features like cell pyknosis and dense granule in nucleus were significantly clearer. More cells were stained with DAB after treated with F200. The flow cytometry results showed 0.71μM F200 could induce 32.6%cell apoptosis. Moreover, the Western Blot results showed cleaved PARP was increased along with the increase of F200 concentration. Conclusion CDK inhibitor F200 could inhibit MCF7 cell growth and induce the cell apoptosis.
