摘要
目的探讨核因子-κB(NF-κB)抑制剂吡咯烷二硫代氨基甲酸(PDTC)对顺铂抑制胃癌细胞SGC-7901增殖的影响。方法不同浓度的PDTC(25,50,75μmol/L)联合顺铂(4mg/L)作用于胃癌细胞SGC-7901,采用细胞增殖/毒性检测试剂盒(CCK-8试剂盒)检测细胞增殖抑制率;实时荧光定量多聚酶链反应(RTPCR)检测NF-κB p65 mRNA的表达情况;免疫荧光细胞化学(IF)检测NF-κB p65蛋白的表达变化。结果CCK-8结果显示,SGC-7901细胞增殖抑制率随PDTC浓度的增加而增加,且高于单独顺铂组,差异有统计学意义。RT-PCR结果显示,与对照组比较,顺铂(4mg/L)能增加p65 mRNA的表达,差异有统计学意义;当与不同浓度PDTC(25,50,75μmol/L)联合作用时,p65 mRNA相对表达量呈剂量依赖性降低,且低于顺铂单独组。免疫荧光结果与RT-PCR结果一致,与对照组比较,顺铂(4mg/L)能增加NF-κB p65蛋白的表达;与PDTC联合时,随PDTC浓度的增加,p65蛋白的表达量降低,积分光密度(IOD)逐渐降低,且低于顺铂单独组。结论 NF-κB抑制剂PDTC能增加顺铂对胃癌细胞SGC-7901化疗的敏感性,提示NF-κB途径是胃癌治疗的重要靶点。
Objective To investigate the effect of the nuclear factor-KB inhibitor pyrrolidine dithiocarbamate （ PDTC ） on the pro- liferation of human gastric cancer cell SGC-7901 inhibited by cisplatin. Methods Different concentrations of PDTC （25,50,751mol/L） in combination with eisplatin（4mg/L） were used in gastric cancer cell SGC-7901. The inhibition rate of gastric cancer cell proliferation was de- termined by cell proliferation/toxicity testing kits （ CCK-8 ） assay. The mRNA expressions of NF-κB p65 were determined by Real-Time poly- merase chain reaction. The changes of NF-κB p65 protein were observed by immunofluoreseence assay. Results CCK-8 showed that the cell proliferation inhibition rate increased along with the increasing of PDTC concentration, and higher than the cisplatin single group, the differ- ence was statistically significant. RT-PCR showed that cisplatin（4mg/L） could increase the p65 mRNA expression compared with the control group ,and the difference was statistically significant. When cisplatin combined with different concentrations of PDTC（25,50,75μmol/L） ,the p65 mRNA expression were reduced in a dose-dependent manner,and lower than that in the cisplatin single group. The results of IF were in agreement with RT-PCR. Compared with the control group,cisplatin（4mg/L） could increase the p65 mRNA expression. When combined with PDTC（25,50,75μmol/L） ,with the increase of PDTC, the 1365 protein expression and integral optical density（IOD） were reduced, and lower than the cisplatin single group. Conclusion NF-κB inhibitor PDTC could increase the sensitivity of cisplatin in the treatment of gastric canc- er cell SGC-7901 ,suggesting that NF-κB pathway is an important target in the treatment of gastric cancer.
作者
王燕泽
崔璨
张红梅
张丽
焦建新
高志星
季万胜
WANG Yan-ze CUI Can ZHANG Hong-mei ZHANG Li JIAO Jian-xin GAO Zhi-xing JI Wan-sheng(Department of Internal Medicine, Weifang Medical University, Weifang 261053, China Department of Gastroenterology, the Affiliated Hospital of Weifang Medical University)
出处
《潍坊医学院学报》
2016年第4期297-300,共4页
Acta Academiae Medicinae Weifang
基金
山东省优秀中青年科学家科研奖励基金资助项目(项目编号:BS2010SW034)
