NF-κB亚单位p50/p65激活促进肺腺癌H1650细胞吉非替尼耐药  

Activation of nuclear factor-κB subunit p50/p65 enhances gefitinib resistance of lung adenocarcinoma H1650 cell line

作　　者：潘莹 黄思超 王霞 龚五星 梁翠微 杜均祥 彭东旭 谢云 郑礼平 张楠 全文 PAN Ying;HUANG Sichao;WANG Xia;GONG Wuxing;LIANG Cuiwei;DU Junxiang;PENG Dongxu;XIE Yun;ZHENG Liping;ZHANG Nan;QUAN Wen(Department of Oncolog;Department of Pharmacy, Zhuhai People＇s Hospital, Zhuhai 519000, China;population and Family Planning Service Center of Zhuhai, Zhuhai 519000, China)

机构地区：[1]珠海市人民医院肿瘤科,广东珠海519000 [2]珠海市人民医院药学部,广东珠海519000 [3]珠海市人口和计划生育服务中心,广东珠海519000

出　　处：《南方医科大学学报》2018年第5期584-590,共7页Journal of Southern Medical University

基　　金：珠海市科技计划医疗卫生项目(一般项目)(20161027E030026)

摘　　要：目的探究NF-κB经典通路激活与人肺腺癌H1650细胞吉非替尼耐药的内在联系。方法选择肺腺癌HCC827和H1650亲代细胞株,吉非替尼连续处理60 d,检测细胞株生长情况和各细胞株595nm吸光度,确定各细胞株细胞存活率和H1650亲代细胞被诱导成H1650耐药细胞株。Western blot检测各细胞株P-IκBα、NF-κB P-p50和P-p65表达量,Image J灰度仪计算出胞浆和胞核P-p50和P-p65与内参蛋白灰度比值,比较吉非替尼和吉非替尼加吡咯烷二硫代氨基甲酸盐(PDTC)联合用药状态下,各细胞株细胞存活率和P-IκB、P-p50和P-p65的表达。结果在无吉非替尼和吉非替尼加PDTC药物干预下,H1650耐药细胞PIκBα表达增加,胞浆和胞核P-p50和P-p65表达,H1650耐药细胞株>H1650亲本细胞株>吉非替尼敏感HCC827细胞株(P<0.05,P<0.01);单独使用吉非替尼干预,H1650耐药细胞株细胞抑制率较H1650亲本细胞株和HCC827细胞株降低(P<0.05,P<0.01),H1650耐药细胞株P-p50和P-p65表达较其他两类细胞株升高(P<0.05);与无吉非替尼干预比较,H1650亲代和耐药细胞P-p50和P-p65均有显著降低(P<0.05,P<0.01);吉非替尼联合PDTC干预,显示H1650亲代和耐药细胞存活率、胞浆和胞核Pp50和P-p65表达均较吉非替尼组降低(P<0.01,P<0.01)。结论 NF-κB经典传导通路激活是H1650亲代细胞残存和转化为吉非替尼耐药细胞的主要因素之一,吉非替尼联合PDTC用药可抑制P-IκBα生成和P-p50、P-p65的激活,从而降低H1650残存细胞生存率和耐药细胞的产生。Objective To explore the intrinsic connection between activation of classical nuclear factor-κB（NF-κB） pathway and gefitinib resistance in human lung adenocarcinoma H1650 cells. Methods Human lung adenocarcinoma H1650 cells were exposed to gefitinib continuously for 60 days to obtain resistant H1650 cells. The expressions of P-IκBα, P-p50 and P-p65 in the cytoplasm or nuclei were detected using Western blotting in human lung adenocarcinoma HCC827 cells, parental H1650 cells and gefitinib-resistant H1650 cells. The effects of gefitinib alone or in combination with PDTC on the survival rate and expressions of NF-κB P-p50 and P-p65 were compared among the 3 cell lines. Results Gefitinib-resistant H1650 cells showed increased cytoplasmic and nuclear P-IκBα expressions. The expressions of P-p50 and P-p65 differed significantly among the 3 cell line, decreasing in the order of resistant H1650 cells, parental H1650 cells, and gefitinib sensitive HCC827 cell lines（P〈0.05 or 0.01）. Treatment with gefitinib alone resulted in a significantly lower cell inhibition rate in resistant H1650 cells than in the parental H1650 cells（P〈0.05） and HCC827 cells（P〈0.01）. The resistant H1650 cells had a significantly higher expression of P-p50 and P-p65 than other two cell lines（P〈0.05）. In both the resistant and parental H1650 cells, gefitinib significantly lowered P-p50 and P-p65 expressions（P〈0.05 or 0.01）, and the combined treatment with gefitinib and PDTC significantly decreased the cell survival rate and further lowered the cytoplasmic and nuclear expressions of P-p50 and P-p65（P〈0.01 or 0.01）. Conclusion The activation of classical NF-κB pathway is a key factor contributing to transformation of the parental H1650 cells into gefitinib-resistant cells. Gefitinib combined with PDTC can inhibit P-IκBα production and NF-κB P-p50 and P-p65 activation to suppress the survival of residual H1650 cells and the generation of gefitinib-resistant cells.

关 键 词：肺腺癌 活化的B细胞核因子κ-轻链增强子 磷酸化p50和p65 H1650细胞株 吡咯烷二硫代氨基甲酸盐 

分 类 号：R734.2[医药卫生—肿瘤]