硒化卡拉胶联合阿霉素对HepG-2/ADR细胞的协同抗肿瘤效应及逆转机制研究  

Synergistic antitumor effect and reversal mechanism ofκ-selenocarrageenan in combination with adriamycin on HepG-2/ADR cells

作　　者：田海燕 凌娜 高铭泽[1,2] 徐贵国 郭春秋 TIAN Haiyan;LING Na;GAO Mingze;XU Guiguo;GUO Chunqiu(Pharmaceutical Engineering Technology Research Center,Harbin University of Commerce,Harbin 150076,China;Engineering Research Center of Anti-Tumor Natural Drugs,Ministry of Education,Harbin 150076,China)

机构地区：[1]哈尔滨商业大学药物工程技术研究中心,黑龙江哈尔滨150076 [2]国家教育部抗肿瘤天然药物工程研究中心,黑龙江哈尔滨150076

出　　处：《中国海洋药物》2024年第3期29-36,共8页Chinese Journal of Marine Drugs

基　　金：黑龙江省双一流学科协同创新成果项目(LJGXCG2023-039);黑龙江省中医药科研项目(ZYW2022-093);黑龙江省省属本科高校基本科研业务费项目(2020CX38,2023-KYYWF-1088)资助。

摘　　要：目的 探究硒化卡拉胶(KSC)联合阿霉素(ADR)对肝癌耐药细胞HepG-2/ADR的协同抗肿瘤效应及其逆转作用机制。方法 MTT法分别检测HepG-2/ADR耐药细胞对ADR、顺铂(DDP)、紫杉醇(TAX)3种化疗药物的多药耐药性,并判断KSC对HepG-2/ADR耐药细胞的逆转作用;流式细胞术检测KSC和ADR对HepG-2/ADR耐药细胞周期和细胞凋亡的影响;Western blot检测KSC和ADR对HepG-2/ADR细胞中耐药相关蛋白、细胞周期和细胞凋亡蛋白的影响。RT-qPCR进一步验证HepG-2/ADR细胞中耐药相关基因的表达。结果MTT结果显示,HepG-2/ADR耐药细胞对ADR、TAX、DDP均具有耐药性,且KSC具有逆转肝癌HepG-2/ADR细胞多药耐药性的作用,两药联合表现为相加作用。流式细胞术表明KSC和ADR均可诱导HepG-2/ADR细胞发生凋亡和S期周期阻滞。Western blot结果显示,KSC和ADR显著下调耐药蛋白P-gp、MRP1、ABCG2以及细胞周期蛋白Cyclin A、Cyclin E、CDK2、Survivin和抑凋亡因子Bcl-2的表达(P<0.05或P<0.01),显著上调促凋亡蛋白Bax、Caspase-3、Caspase-9、Cleaved-Caspase-3和Cleaved-Caspase-9的表达(P<0.05或P<0.01);联合组可协同促进或抑制细胞凋亡和细胞周期相关蛋白的表达。RT-qPCR表明KSC和ADR单独及联合应用均能显著下调P-gp、MRP1和ABCG2耐药基因的表达。结论 KSC可有效逆转肝癌多药耐药性,协同提高HepG-2/ADR细胞对ADR的化疗敏感性,其机制可能与诱导细胞凋亡和细胞周期阻滞有关,通过下调多药耐药相关蛋白P-gp、MRP1和ABCG2的表达,进而逆转肝癌多药耐药性。Objective To explore the synergistic antitumor effect and reversal mechanism ofκ-selenocarrageenan(KSC)in combination with adriamycin(ADR)on hepatoma resistant cell line HepG-2/ADR.Methods The multidrug resistance of HepG-2/ADR resistant cells to adriamycin(ADR),cisplatin(DDP)and paclitaxel(TAX)was detected by MTT assay,in order to evaluate the reversal effect of KSC on HepG-2/ADR cells.The effects of KSC and ADR on the cell cycle and apoptosis of HepG-2/ADR cells were detected by flow cytometry.Western blot was used to detect the expressions of drug-resistant proteins,cyclins and apoptosis-related proteins in HepG-2/ADR cells exposed to KSC and ADR.Moreover,the expressions of drug resistance related genes in HepG-2/ADR cells were further confirmed by RT-qPCR.Results MTT results showed HepG-2/ADR cells were resistant to ADR,TAX and DDP,demonstrating multi-drug resistance.KSC could reverse the multidrug resistance of HepG-2/ADR cells,and exhibited an additive effect while in combined with ADR on HepG-2/ADR cells.Flow cytometry indicated both KSC and ADR could induce apoptosis and S phase arrest of HepG-2/ADR cells.Additionally,western blot analysis showed KSC and ADR significantly downregulated the expressions of multidrug-resistant protein P-gp,MRP1 and ABCG2,as well as Cyclin A,Cyclin E,CDK2,Survivin and anti-apoptotic protein Bcl-2(P<0.05 or P<0.01),and significantly upregulated the expressions of pro-apoptotic proteins such as Bax,Caspase-3,Caspase-9,Cleaved-Caspase-3,and Cleaved-Caspase-9(P<0.05 or P<0.01).Besides,the combined group could synergistically promote or inhibit the expressions of apoptosis and cell cycle-related proteins.Furthermore,RT-qPCR demonstrated KSC and ADR alone or in combination could significantly downregulate the mRNA expression of P-gp,MRP1 and ABCG2.Conclusion KSC could effectively reverse multidrug resistance of hepatoma and synergistically enhance the sensitivity of HepG-2/ADR cells to adriamycin.The mechanism may be related to the induction of apoptosis and cell cycle arrest,as w

关 键 词：硒化卡拉胶 阿霉素 肝癌耐药细胞HepG2/ADM 多药耐药性 细胞凋亡 

分 类 号：R931.77[医药卫生—生药学] R96[医药卫生—药学]