摘要
目的 探讨去甲斑蝥素 (NCTD)对人胆囊癌GBC -SD细胞系生长的影响及其机制。方法 实验分NCTD组 ( 7个浓度梯度组 ,每组 6孔 )和对照组 (n =6 ) ,分别应用MTT、流式细胞术、光学和电子显微镜、SABC法检测NCTD对体外培养GBC -SD细胞的杀伤抑制率、细胞周期和凋亡、细胞形态学改变和增殖相关基因蛋白PCNA、Ki- 6 7表达。结果 NCTD在浓度 10μg/ml、时间 6h时即对GBC -SD细胞生长有抑制作用 ,随浓度升高、时间延长作用增强 ,呈剂量 -时间效应关系。在实验组 ,流式细胞仪示GBC -SD细胞的G2 +M期细胞明显增多 ,S期细胞减少 ,凋亡率上升 ;光镜下出现细胞固缩 ,胞膜突出 ,核碎裂和凋亡小体 ;电镜示微绒毛卷缩、高尔基体、线粒体等细胞器衰退现象和典型凋亡细胞 ;SABC法检测示PCNA、Ki- 6 7表达阳性的细胞数和阳性指数明显少于对照组 (PCNA :0 .932± 0 .0 31vs 0 .318± 0 .0 2 3;Ki- 6 7:0 .96 4± 0 .0 92vs 0 .2 97± 0 .0 18;P <0 .0 0 1)。结论 NCTD能抑制人原发性胆囊癌GBC -SD细胞的生长 ;其机制可能与其抑制GBC -SD细胞增殖、干扰生长周期、抑制DNA合成代谢、诱导细胞凋亡和影响细胞增殖相关基因蛋白PCNA、Ki- 6
Objective To study the influence of norcantharidin (NCTD) on growth of GBC-SD cell line with human gallbladder carcinoma and its mechanism. Methods GBC-SD cells for human gallbladder carcinoma were cultured by cell culture technique. The experiment was divided into NCTD group and control group. The tetrazolium-based colorimetric assay was used to evaluate the inhibitory effect of NCTD for growth of GBC-SD cells. Flow cytometry was used to measure the induction of cell cycle arrest and apoptosis. Microscope and scan electron microscope were used to observe the morphological changes of the cells. Results NCTD showed the inhibitory effect on growth of GBC-SD cells from 10 μg/ml after 6 hours. It was in a dose- and time-effective dependent manner. The flow cytometric profiles revealed that NCTD led to the shift from 28.54% up to 58.71% of the G 2/M phase and from 24.47% down to 6.33% of the S phase cells in percentage of GBC-SD cells, with the increased rate of cell apoptosis. In NCTD group, cell shrinkage, membrane budding, and karyorrhexis of GBC-SD cells could been seen by microscope; microvillus decreasing, atrophy of cell apparatus including Golgi and mitochondria as well as typical apoptosis cells could also been seen by electron microscope; the expressions and their positive index (PCNA: 0.932 vs 0.318, Ki-67: 0.964 vs 0.297; P <0.001) of PCNA, Ki-67 gene proteins of GBC-SD cells were decreased significantly. Conclusion NCTD can inhibit the growth of GBC-SD cell lines with human gallbladder carcinoma. The mechanism might correlate with inhibition of cell proliferation, blockade of cell cycle, inhibition of DNA synthesis and metabolism, induction of cell apoptosis and inhibition of PCNA and Ki-67 expressions in GBC-SD cells.
出处
《肿瘤》
CAS
CSCD
北大核心
2004年第4期358-361,共4页
Tumor
