摘要
目的 :探讨ATP及其衍生物对不死化人成纤维细胞的增殖抑制作用 ,并通过P2 嘌呤能受体从而了解ATP发挥细胞毒性作用的途径。方法 :使用ATP及其衍生物ATP -Na2 ,ATP -Mg ,ADP ,MeATP ,BzATP ,ATPγS ,2 -MeSATP和UTP在不同条件培养KMST - 6人不死化成纤维细胞 ,检测细胞增殖状况 ,Westernblot分析、流式细胞仪检测细胞增殖周期以及Hoechst 3 3 2 5 8特异性细胞染色和DNA电泳检测细胞凋亡。结果 :ATP及其衍生物对细胞增殖抑制作用的程度依次为 :ATP =ADP >ATPγS >MeATP =BzATP ;而 2 -MeSATP和UTP却未显示任何细胞毒性作用 ;0 4mmol/LATP培养 4 8h时P2 1表达未见增高 ,细胞增殖停止于G1/S期 ;1mmol/LATP培养 4 8h时未发现细胞凋亡。结论 :通过与P2X或P2Y嘌呤能受体相结合 ,ATP激活细胞内某些信号传导发挥细胞增殖的抑制作用 ;ATP引起增殖抑制不是通过细胞凋亡或者是周期素 /CDK激酶抑制剂P2 1所致。
AIM: To investigate the inhibitory effects of ATP on proliferation of immortalized human fibroblasts, and learn what subtypes of P-2 purinergic receptors are involved in ATP cytotoxicity. METHODS:The immortalized human fibroblast cell line KMST-6 was cultured with ATP and its derivatives including ADP, β,γ-methyleneadenosine5'-triphosphate(MeATP), 2'&3'-O-(4-benzoylbenzoyl) adenosine, triethylammonium salt (BzATP), adenosine 5'-O-(3-methiotriphosphate)(ATPγS), 2-methylthioadenosine 5'-triphosphate(2-MeATP) and UTP, and the cell cycle analysis was finished using flow-cytometer, the cell apoptosis analysis was made using Hoechst 33258 and examination of DNA ladder. RESULTS: The extent of cytotoxicity induced by these drugs was found to be in order of : ATP=ADP>ATPγS>MeATP=BzATP, neither 2-MeSATP nor UTP showed any cytotoxicity. No enhanced expression of P 21 was observed and the cell cycle was held in G 1/S in the cells treated with 0.4mmol/L;no cell apoptosis was detected in the cells treated with 1mmol/L ATP. CONCLUSION: Connecting with P 2X or P 2Y purinergic receptors,ATP activated some intracellular signals to inhibit cell growth, the growth inhibition caused by ATP was not due to apoptosis or induction of cyclin/CDK kinase inhibitor,P 21 .
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2001年第2期150-153,共4页
Chinese Journal of Pathophysiology
关键词
腺苷三磷酸
细胞
受体
嘌呤能
Adenosine triphosphate
Cells
Receptors,purinergic
