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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 ,可用于大批量抗肿瘤药物的筛选
Objective To establish a simple and reliable method for screening anti-tumor cell growth and apoptosis-inducing drugs. Methods Based on the close relationship between acid phosphatase activity and cell proliferation in the cytoplasm, the correlation between cell number and acid phosphatase activity was determined. A 96-well micro-measurement method was established. Flow cytometry was used to detect cells at the same time. Cycle and apoptosis, determine the relationship between apoptotic cells and acid phosphatase, and compare sensitivity with thiazolyl (MTT) method. RESULTS: In two hepatocellular carcinoma cell lines (HepG2 cells and CBRH 7919 cells), the activity of acid phosphatase was positively correlated with the number of cells, and was linear in the range of (0.5 - 7) x 10 3 cells. The correlation coefficient (CV) was 0.994; after 1h of phorbol ester (TPA)-stimulated hepatoma cells, the activity of acid phosphatase increased significantly. On the other hand, after the action of arsenic trioxide, the activity of acid phosphatase decreased significantly, and the higher the concentration, the more significant the decline. The arsenic trioxide exposed HepG2 cells for 24 hours, and the apoptotic cells accounted for only 3.98%. The effect of CBRH 7919 cells on the acid phosphatase activity was significantly decreased when no apoptosis occurred. Conclusion The detection of acid phosphatase activity in cells can be used as an indicator of cell proliferation and apoptosis. This method is simple and rapid and can be used for the screening of large-volume antitumor drugs.