摘要
背景与目的:活化的核转录因子-kappa B(nuclear factor-kappa B,NF-κB)信号通路在多种肿瘤的发生、发展中起着重要的作用,参与调控多种与炎症、抗凋亡、肿瘤形成和转化有关基因的表达。目前其在食管鳞癌中的作用尚不清楚。本研究旨在探讨p65siRNA和姜黄素能否通过阻断NF-κB信号通路,促进食管鳞癌细胞的凋亡及增强对化疗药的敏感性,并将两种方法加以比较。方法:Western blot检测p65siRNA转染到食管鳞癌细胞EC9706和Eca109中p65蛋白的表达;EC9706和Eca109细胞中加入姜黄素(50μmol/L)后检测pIκBα蛋白的表达。转染p65siRNA的食管鳞癌细胞,单独或联合使用氟尿嘧啶(5-fluorouracil,5-FU)治疗72 h;EC9706和Eca109细胞中加入姜黄素培养72 h,或联合使用姜黄素和5-FU培养72 h,流式细胞仪检测各组细胞凋亡情况。显微镜下观察各组食管鳞癌细胞形态学特性的改变。结果:Western blot结果显示,转染p65siRNA的EC9706和Eca109细胞中,p65蛋白的表达水平下调,而非靶向蛋白MAPK的表达无影响;姜黄素组中,EC9706和Eca109细胞中pIκBα蛋白的表达水平随着姜黄素作用时间的延长逐渐下降。将siRNA转染组与姜黄素组所引起的食管鳞癌细胞的凋亡及联合应用化疗药所引起的细胞凋亡加以比较,结果发现,与siRNA转染组相比,单独使用姜黄素可引起细胞凋亡数增加(P<0.05);但姜黄素与5-FU联合应用时,可明显提高肿瘤细胞对化疗药的敏感性,凋亡和坏死的细胞均显著增加(P<0.05)。结论:p65siRNA和姜黄素都可以通过阻断NF-κB信号通路,促进食管鳞癌细胞的凋亡及增强对5-FU的敏感性;RNA干扰虽特异,但用于临床还需要一段时间。姜黄素不良反应小,有望成为治疗食管癌的辅助用药。
Background and purpose:The activation of NF-κB signaling pathway plays a critical role in the initiation and progression of carcinogenesis.Although constitutive NF-κB activation has been reported in many human tumors,the role of the NF-κB pathway in esophageal squamous cell carcinoma(ESCC) is not clear.The purpose of this study was to detect whether p65siRNA and curcumin could promote ESCC cells apoptosis as well as to increase the ESCC cells' sensitivity to chemotherapeutic drugs by inhibiting the NF-κB signaling pathway.These 2 outcomes will be compared.Methods:The expression of p65 was detected in ESCC cells transfected with p65siRNA by Western blot.After being treated with curcumin,ESCC cells were examined for the expression of pIκBα using Western blot.After ESCC cells were treated with p65siRNA and curcumin alone or combined with 5-FU for 72 h,the cells were analyzed for cell apoptosis using flow cytometry.Morphological changes of ESCC cells were observed under microscope.Results:Western blot results indicate that the protein level of p65 decreased after being transfected under p65siRNA,while the protein level of MARK was not affected.Curcumin seemed to have inhibited IκBα phosphorylation and degradation in the 2 ESCC cell lines in a time-dependent manner.Compared with p65siRNA,curcumin alone could increase cell apoptosis(P<0.05),but when combined with 5-FU,the results were more prominent(P<0.05).The proliferation of EC9706 and Eca109 was slow after being treated with p65siRNA and curcumin in combination with 5-FU.Conclusion:Both p65siRNA and curcumin could promote ESCC cells apoptosis and enhance sensitivity to 5-FU through suppression of the NF-κB signaling pathway.There is still a long way for RNA interference practicing in clinic.Therefore,curcumin is proved to be useful in the treatment of ESCC because it is a pharmacologically safe compound with less side effects.
出处
《中国癌症杂志》
CAS
CSCD
北大核心
2011年第5期326-331,共6页
China Oncology
基金
河南省医学科技攻关项目(No:200803005)
郑州市科技攻关项目(No:0910SGYS33389-7)
