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细梗香草皂苷联合吉西他滨对胰腺癌细胞BxPC-3抑制作用的研究
于贤金, 何亚红, 张筱凤
杭州市第一人民医院消化内科
摘要:
目的观察细梗香草皂苷(LC)和吉西他滨(GEM)单药及联合用药对人胰腺癌BxPC-3细胞增殖和凋亡的影响,探究LC抗胰腺癌的作用机制。方法培养人胰腺癌BxPC-3细胞时加入不同浓度梯度的LC与GEM,采用MTS法检测细胞增殖抑制率,计算两种药物的半抑制浓度(IC50)和联合作用指数(CI)。将细胞分为4组,A组(对照空白)、B组(GEM1滋mol/L)、C组(LC15滋g/ml)和D组(GEM1滋mol/L+LC15滋g/ml联合用药),培养后采用AnnexinⅤ-FITC/PI法观察LC与GEM对BxPC-3细胞的诱导凋亡作用;Westernblot法检测抗多聚(ADP-核糖)聚合酶(PARP)、抗半胱氨酸蛋白酶-3(Caspase-3)的表达。结果8~64滋g/ml的LC及1.25~20滋mol/L的GEM均对BxPC-3细胞的增殖有抑制作用,且呈浓度依赖性,其IC50分别为16.55滋g/ml和1.27滋mol/L。15滋g/ml的LC与各个浓度的的GEM协同作用最强,CI值为0.53~0.65。B、C、D组的早、晚期凋亡率与A组比较差异均有统计学意义(均P<0.01),且D组与B、C组的早期凋亡率比较差异均有统计学意义(均P<0.01)。C、D组与A组比较,PARP蛋白表达水平明显降低(均P<0.05);B、C、D组与A组比较,Caspase-3蛋白表达水平增加(均P<0.05)。结论LC有抑制胰腺癌细胞生长的作用,与GEM联合作用效果更好;其作用机制可能是通过激活Caspase-3表达,分解PARP,从而诱导细胞凋亡。
关键词:  胰腺癌 细梗香草皂苷 吉西他滨 PARP Caspase-3 细胞凋亡
DOI:10.12056/j.issn.1006-2785.2018.40.2.2017-920
分类号:
基金项目:浙江省中医药科技计划项目(2013ZZ012)
Effect of lysimachia capillipes with gemcitabine on proliferation and apoptosis of pancreatic cancer BxPC-3 cells and related mechanism
Hangzhou First People's Hospital
Abstract:
Objective To investigate the effect of lysimachia capillipes(LC) and gemcitabine(GEM) on proliferation and apoptosis of human pancreatic cancer cell line BxPC-3 in vitro. Methods The cultured BxPC-3 cells were treated with GEM (1滋mol/L, group B), LC (15滋g/ml, group C) and GEM with LC (15滋g/ml and 1滋mol/L, group D) for 48h, respectively. The untreated BxPC-3 cells served as controls (group A). The proliferation of BxPC-3 cells was measured by MTS method, half maximal inhibitory concentration(IC50) and combined index(CI) of the two drugs were analyzed with Calcusyn software. Cell apoptosis was examined by using flow cytometry(FCM) with AnnexinⅤ/PI staining solution. The expression of proteins PARP and Capase-3 was detected by Western blot. Results The proliferation of BxPC-3 cells was inhibited by LC and GEM. The IC50 value of LC and GEM were 16.55滋g/ml and 1.27滋mol/L, the CI value was 0.53-0.65, when BxPC-3 cells treated with 15-20滋g/ml LC and 1-30滋mol/L GEM. The apoptosis rate of BxPC-3 cells was increased in group B, C and D compared with group A (all P<0.01), and the effect of group D is stronger than that of group B and C (both P<0.01). The expression of Capase-3 protein was up-regulated in group B, C and D. Compared with A (all P<0.05), and the expression of PARP protein was down-regulated in group C and D Cormpared with A(both P<0.05). Conclusion LC and GEM can inhibit the proliferation of BxPC-3 cells, and the effect of combination of the two drugs is more marked. LC can induce apoptosis of BxPC-3 cells by activating the Caspase-3 protein expression and decomposing PARP protein.
Key words:  Pancreatic cancer Lysimachia capillipes Gemcitabine PARP Capase-3 Cell apoptosis