酸性磷酸酶法检测肿瘤细胞的增殖状态  

Acid phosphatase assay for measuring the proliferation of tumor cells

作　　者：卢虹[1] 吴兴中[1] 

机构地区：[1]复旦大学基础医学院生化教研室，上海 200032

出　　处：《中华检验医学杂志》2001年第2期84-86,共3页Chinese Journal of Laboratory Medicine

基　　金：上海市教委重点学科基金项目(B980801)

摘　　要：目的　建立一种简便可靠的筛选抗肿瘤细胞生长和诱导凋亡药物的方法。方法　以细胞浆中酸性磷酸酶活力和细胞增殖之间具有紧密关系为依据 ,确定细胞数目与酸性磷酸酶活力之间的相关性 ,建立 96孔板微量测定方法 ,同时运用流式细胞仪检测细胞周期和凋亡 ,确定凋亡细胞与酸性磷酸酶的关系 ,并与噻唑兰 (MTT)方法进行灵敏度的比较。结果　在 2种肝癌细胞株 (HepG2 细胞和CBRH 7919细胞 )中 ,酸性磷酸酶的活力与细胞数目呈正比关系 ,在 (0 .5～ 7)× 10 3 细胞数范围内呈直线关系 ,二者相关系数 (CV)达 0 .994;在佛波酯 (TPA)刺激肝癌细胞 1h后 ,酸性磷酸酶的活力即显著上升。相反在三氧化二砷作用后 ,酸性磷酸酶的活力即显著下降 ,浓度越高 ,下降越显著。三氧化二砷作用HepG2 细胞 2 4h ,凋亡细胞仅占 3.98% ,作用CBRH 7919细胞 ,在还没有出现凋亡时 ,酸性磷酸酶的活力即已显著下降。结论　检测细胞的酸性磷酸酶活力可作为细胞增殖和凋亡的指标 ,此方法简便、快速 。Objective To establish a simple assay for quick scanning of effective agents in growth inhibition and apoptosis induction. Methods After observing the correlation between cell number and acid phosphatase activity, and between cell apoptosis and acid phosphatase activity, we established a microplate densimetry method to measure the acid phosphatase activity. We also compared the sensitivity with MTT assay and confirmed cell apoptosis by flow cytometry. Results In two cell lines of cancer (Hep G 2 and CBRH 7919 cells), the acid phosphatase activity was correlated well with the cell number, and was directly proportional to the cell number in the range of 0.5×10 3～0.7×10 3. The coefficient reached to 0.994. After stimulation of phorbel ester (TPA) for one hour, the acid phosphatase activity rose significantly. In contrast, after the inhibition of cell growth by As 2O 3, the acid phosphatase activity decreased significantly. The higher the concentration of As 2O 3, the lower the acid phophatase activity. The acid phophatase activity went down significantly when apoptosis in Hep G 2 cells was only 3.98% and no apoptosis in CBRH 7919 cells after 24 hour treatment with As 2O 3. Conclusions The acid phophatase activity of cell may be used to measure cell proliferation and apoptosis. Because of the easiness and quickness, acid phophatase assay might be an ideal approach to scan the effective growth inhibitors and apoptosis inducers in a large number of drugs.

关 键 词：酸性磷酸酶 肿瘤细胞 细胞培养 流式细胞术 抗癌药 细胞周期 药物筛选 细胞增殖 

分 类 号：R730.4[医药卫生—肿瘤]