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石丹,马旖旎.汉防己甲素体外辅助多柔比星杀伤胃癌细胞及机制研究[J].浙江中西医结合杂志,2018,28(11):
汉防己甲素体外辅助多柔比星杀伤胃癌细胞及机制研究
Effect and mechanical research of tetrandrine on promoting doxorubicin-induced cytotoxicity against gastric cancer in vitro
投稿时间:2018-04-25  修订日期:2018-09-27
DOI:
中文关键词:  汉防己甲素  caveolin-1  多柔比星  胃癌  凋亡
英文关键词:tetrandrine  caveolin-1  doxorubicin  gastric cancer  apoptosis
基金项目:
作者单位E-mail
石丹* 西溪街道社区卫生服务中心 myt22@163.com 
马旖旎 杭州甄元健康科技有限公司  
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中文摘要:
      目的 探讨中药活性成分汉防己甲素是否能辅助多柔比星杀伤胃癌细胞并研究其机制。方法: 胃癌细胞系MGC-803的相对细胞活力用MTT法进行检测。MGC-803细胞中caveolin-1的表达水平,AKT和Bad的磷酸化水平,caspase-9和caspase-3的活化水平用Western blot实验进行检测。Bad与Bcl-xl及Bcl-2的相互作用用免疫共沉淀法进行检测。MGC-803细胞的凋亡用流式细胞术进行检测。结果: 汉防己甲素能增强多柔比星的抗胃癌活性,汉防己甲素联合多柔比星组MGC-803的相对细胞活力(0.34±0.03)显著低于多柔比星单处理组(0.79±0.06)(P<0.05),而汉防己甲素联合多柔比星组MGC-803的细胞凋亡率(40.3±3.5)%显著高于多柔比星单处理组(12.2±1.1)%(P<0.05)。汉防己甲素联合多柔比星组MGC-803细胞的小窝蛋白-1(caveolin-1)表达水平,AKT和Bad的磷酸化水平均低于多柔比星组,而汉防己甲素联合多柔比星组MGC-803细胞的Bad与Bcl-xl及Bcl-2的结合水平,caspase-9和caspase-3的活化水平均均高于多柔比星组。汉防己甲素发挥对多柔比星的辅助治疗作用依赖于caveolin-1的抑制,汉防己甲素+多柔比星+caveolin-1质粒组MGC-803的相对细胞活力(0.71±0.06)显著高于多柔比星+多柔比星组(0.34±0.03)(P<0.05),而其细胞凋亡率(15.9±1.2)%显著低于汉防己甲素+多柔比星组(40.3±3.5)%(P<0.05)。结论: 汉防己甲素通过caveolin-1/AKT/Bad途径增强多柔比星对胃癌细胞的凋亡诱导活性。
英文摘要:
      objective To explore the adjuvant effect and mechanisms of tetrandrine on enhancing the doxorubicin-induced cytotoxicity against gastric cancer in vitro. Methods MTT assay was performed to detect the relative cell viability of MGC-803 cells. Western blot analysis was conducted to detect the expression of caveolin-1, phosphorylation of AKT and Bad, and activation of caspase-9 and caspase-3. Co-immunoprecipitation assay was performed to evaluate the interaction with Bad and Bcl-2/Bcl-xl. Flow cytometry analysis was performed to detect the apoptosis of MGC-803. Results Tetrandrine enhanced the anti-tumor activity of doxorubicin against gastric cancer in vtiro. Relative cell viability of MGC-803 in tetrandrine plus doxorubicin group (0.34±0.03) was significantly lower than the doxorubicin group (0.79±0.06)(P<0.05), whereas the apoptotic rate of MGC-803 in tetrandrine plus doxorubicin group (40.3±3.5) was significantly higher than the doxorubicin group (12.2±1.1)(P<0.05). Expression of caveolin-1 and phosphorylation of AKT and Bad in was tetrandrine plus doxorubicin group was lower than the doxorubicin group, whereas the interaction of Bad with Bcl-xl/Bcl-2, and activation of caspase-9 and caspase-3 was higher than the doxorubicin group. Adjuvant effect of tetrandrine on doxorubicin was dependent on the inhibition of caveolin-1. Relative cell viability of MGC-803 in tetrandrine + doxorubicin + caveolin-1 plasmid group (0.71±0.06) was significantly higher than the tetrandrine plus doxorubicin group (0.34±0.03)(P<0.05), whereas the apoptotic rate of MGC-803 in tetrandrine + doxorubicin + caveolin-1 plasmid group (15.9±1.2) was significantly lower than the tetrandrine plus doxorubicin group (40.3±3.5)(P<0.05). Conclusion tetrandrine enhanced the doxorubicin-induced apoptosis through the caveolin-1/AKT/Bad pathway.
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