摘要
目的:研究金喜素抑制人肺癌SH-77细胞增殖及诱导其凋亡的作用,并探讨其机制。方法:体外培养肺癌细胞,以20mg·L-1金喜素处理细胞24小时后,MTT法检测金喜素对肺癌细胞增殖的影响;TUNEL法和流式细胞仪检测肺癌细胞凋亡情况以及Caspase-3特异性拮抗剂Ac-DEVD-CHO(50μmol·L)对金喜素诱导凋亡作用的影响;放射性同位素法检测肺癌细胞胞膜蛋白激酶C(PKC)活性。结果:金喜素处理细胞24小时后,肺癌细胞的存活率为59.8%±5.5%,未经金喜素处理的对照组细胞存活率为98.3%±9.4%,两组间有显著差异性(P<0.01);TUNEL法观察到典型的凋亡细胞形态学特征,流式细胞仪检测肺癌细胞凋亡率为14.5%±3.1%,显著高于对照组细胞(P<0.01);Ac-EDVD-CHO和金喜素共同处理细胞24小时其凋亡率为7.1%±1.3%,较单纯金喜素作用组明显降低(P<0.01);肺癌细胞膜PKC活性(pmol/mgprotein·min)为45.3±10.7,较对照组(141.8±30.4)显著降低(P<0.01)。结论:金喜素可抑制SH-77肺癌细胞增殖,并通过Caspase-3途径诱导其凋亡而发挥抗肿瘤作用,其作用机制与降低肺癌细胞PKC活性有关。
Objective:To study the effect of topotecan on the proliferation and apoptosis of human lung cancer cells SPC-A-1and its possible mechanism.Methods:Cells were incubat-ed with topotecan20mg.L-1for24h.Cell viability was measured by MTT assay.Morphologi-cal change of apoptotic cells was studied by TUNEL staining.Flow cytometry was used to detect the apoptotic rate of cells incubated with topotecan or topotecan and Ac-DEVD-CHO (50umol-L-1),a specific Capspase-3inhibitor,and protein kinase C(PKC)activity of cell membrane was de-tected.Results:The cell viability with20mg /L TPT for24h was59.8%±5.5%which was high-er than that of cells without topotecan(98.3%±9.4%P<0.01).Typical morphological features of apoptotic cells were detectede by TUNEL staining.apoptotic rate of cells with topotecan was14.5%±3.1%which was higher than that of cells treated with topotecan and Ac-DEVD-CHO(7.1%±1.3%P<0.01).PKC activity(pmol/mg protein.Min)of cells with topotecan was45.310.7which was much lower than that of control cells(141.8±30.4P<0.01).Conclusion:Topotecan can inhibit the cell proliferaton and induce apoptosis of SH-77human lung cancer cells through caspase-3activation.The possible mechanism might be related to the inhibition of PKC activity.
出处
《中国肿瘤临床》
CAS
CSCD
北大核心
2002年第5期361-364,共4页
Chinese Journal of Clinical Oncology
