膀胱癌细胞凋亡检测方法的比较研究

目的 比较几种检测羟基喜树碱（ＨＰＴ）诱导的膀胱癌细胞凋亡的方法。方法 通过流式细胞分析观察ＨＰＴ诱导Ｔ２４膀胱癌细胞凋亡的量效关系;通过荧光染色对凋亡细胞的形态特征进行分析;直接使用苏木素染色进行细胞凋亡的检测;应用ＤＮＡ分析进行ＤＮＡｌａｄｄｅｒ检测。结果 （０．０１￣１０）μｇ／ｍｌ的ＨＰＴ在体外能够诱导Ｔ２４细胞凋亡,其最佳诱导时间在３小时以后。流式细胞（０．０１￣１０）μｇ／ｍｌ的ＨＰ

Detection of apoptosis exposed to 10-hydroxycamptothecin in T24 human urinary bladder cancer cells

OBJECTIVE: To study whether 10-hydroxycamptothecin (HPT) can induce apoptosis in bladder cancer cell and establish methods for detecting apoptotic cells. METHODS: Human urinary bladder cancer cell line (T24) was exposed in vitro to different concentrations of 10-hydroxycamptothein for various lengths of time. Flow cytometry, Hochest 33258 and Hematoxylin staining were used to determine the induction of apoptosis after use of HPT. DNA gel analysis was also carried out to detect DNA fragmentation. RESULTS: Cell shrinage, nuclear fragmentation and condensed chromosomes showed that apoptosis can be induced by HPT within the concentration of 0.01 - 10 microg/ml. The flow cytometry analysis showed that the percentage of apoptotic cells were related to the concentration and the time of induction. T24 cell line exposed to HPT experienced internucleosomal DNA fragmentation by producing a typical ladder pattern on agarose gel electrophoresis. The detection of minimum exposure time for HPT-induced apoptosis in T24 cells showed that 3 hours of exposure to HPT were enough to trigger internucleosomal DNA fragmentation. Compared to Hochest 33258 staining, Hematoxylin staining was more easy, rapid and accurate to detect apoptosis. CONCLUSIONS: The induction of apoptosis exposed to HPT in T24 human urinary bladder cancer cells is a good model for further studying urinary bladder cancer. Hematoxylin staining is a useful method for detecting apoptosis.