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目的 :研究羟喜树碱 (HCPT)体外抑制人胰腺癌细胞株 SW- 1990增殖并诱导其凋亡的作用。方法 :体外培养的胰腺癌细胞经不同质量浓度 (1.5 6 2 5 ,3.12 5 ,6 .2 5 ,12 .5 ,2 5 ,5 0 ,10 0μg/ m l)的 HCPT处理 2 0～ 2 4h后 ,用 MTT法测定细胞增殖情况 ,再分别以 Annexin 早期凋亡检测试剂盒、电子显微镜、流式细胞术和 TU NEL标记以及 bcl- 2免疫细胞化学标记检测细胞凋亡情况。 结果 :(1) 3.12 5～ 10 0μg/ ml HCPT作用 2 4h后 ,肿瘤细胞的增殖抑制率均非常显著高于对照组 (P<0 .0 1)。(2 ) 5 0μg/ m l HCPT作用 2 0 h后 ,经流式细胞术及 TUNEL标记检测 ,肿瘤细胞的凋亡率分别为 13.2 %和 15 .3% ;10 0μg/ m l HCPT作用 2 0 h后 ,两种方法检测的细胞凋亡率分别为 2 6 .1%和 2 8.4%。 (3)电镜及 Annexin 标记荧光检测观察表明 ,5 0μg/ m l HCPT作用下 ,肿瘤细胞主要呈早期凋亡表现 ,凋亡率为 18.4%。 (4 ) 5 0μg/ ml HCPT作用 2 4h后 ,肿瘤细胞bcl- 2表达较对照组明显减弱。 结论 :HCPT在体外可抑制 SW- 1990细胞的增殖 ,部分作用机制是诱导细胞凋亡 ;提示 HCPT可作为细胞凋亡诱导剂用于胰腺癌的治疗。
Objective: To study the effect of hydroxycamptothecin (HCPT) on the proliferation and induction of apoptosis of human pancreatic cancer cell line SW-1990 in vitro. METHODS: Pancreatic cancer cells cultured in vitro were treated with HCPT at different mass concentrations (1.5625, 3.125, 6.5%, 12.5%, 2.5, 50, 100μg/ml) for 20 to 24 hours. The cell proliferation was measured by MTT assay. Apoptosis was detected by Annexin early apoptosis detection kit, electron microscopy, flow cytometry and TUNEL labeling, and bcl-2 immunocytochemistry. RESULTS: (1) After 3.12 5～10 0μg/ml HCPT for 24 hours, the proliferation inhibition rate of tumor cells was significantly higher than that of the control group (P<0.01). (2) After 50 μg/ml HCPT treatment for 20 h, the apoptotic rate of tumor cells was 13.2% and 15.3%, respectively, by flow cytometry and TUNEL labeling; 100 μg/ml HCPT effect was 20 h The rate of cell apoptosis detected by the two methods was 26.1% and 28.4%, respectively. (3) Fluorescence electron microscopy and annexin labeling observation showed that, under the effect of 50 μg/ml HCPT, the tumor cells mainly showed early apoptosis, and the apoptosis rate was 18.4%. (4) After 40 μg/ml HCPT for 24 h, the expression of bcl-2 in tumor cells was significantly decreased compared with the control group. Conclusion: HCPT can inhibit the proliferation of SW-1990 cells in vitro, and part of the mechanism is to induce apoptosis. It is suggested that HCPT can be used as an apoptosis inducer for the treatment of pancreatic cancer.