左金丸调控PI3K/AKT/NF-κB信号通路对人大肠癌干细胞细胞耐药性的影响  

Effects of Zuojin Pill on drug resistance of human colorectal cancer stem cells via PI3K/AKT/NF-κB signaling pathway

作　　者：隋华[1] 卫真真 王子元[2] 孙健 刘宁宁[1] 冯媛媛[1] 刘宣[1] 朱惠蓉[1] 李琦[1] SUI Hua;WEI Zhenzhen;WANG Ziyuan;SUN Jian;LIU Ningning;FENG Yuanyuan;LIU Xuan;ZHU Huirong;LI Qi(Department of Medical Oncology,Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Department of Pathology,Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Clinical Laboratory,Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]上海中医药大学附属曙光医院肿瘤科,上海201203 [2]上海中医药大学附属曙光医院病理科,上海201203 [3]上海中医药大学附属曙光医院检验科,上海201203

出　　处：《上海中医药杂志》2020年第S01期127-131,共5页Shanghai Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(81573805,81874399,81703893,81673783,81874405).

摘　　要：目的探讨左金丸(ZJW)通过PI3K/AKT/NF-κB信号通路对人结肠癌干细胞耐药的调节机制。方法使用无血清培养法从人结肠癌HCT-116细胞中富集得到悬浮肿瘤细胞球,并将其传代扩增;同时在含有奥沙利铂(L-OHP)的培养基常规培养HCT-116/L-OHP耐药细胞。采用Cell Counting Kit-8(CCK-8)法检测ZJW联合L-OHP对细胞化疗耐受性的影响,使用real-time PCR检测ZJW干预肿瘤干细胞后细胞干性标志CD133、Nanog、Oct4、CD44、SOX2等的表达情况;应用Western blot检测ZJW对干细胞和耐药细胞PI3K/AKT/NF-κB信号通路中AKT、p-AKT(Thr308、Ser473)、NF-κB以及p-NF-κB蛋白表达水平的影响。结果ZJW可增加HCT-116/SP细胞对化疗药物的敏感性,并呈一定的量-效关系;ZJW能明显抑制HCT-116/SP细胞CD44和SOX2的mRNA的表达,降低HCT-116/SP和HCT-116/L-OHP细胞的p-AKT和p-IκB的表达水平(P<0.01),并呈现剂量依赖性。结论ZJW可通过降低p-AKT、p-IκB蛋白的磷酸化水平,抑制PI3K/AKT/NF-κB信号通路的激活,降低CD44、SOX2等干性标志的表达,逆转HCT-116/SP细胞的耐药。Objective To investigate the regulation mechanism of Zuojin Pill on drug resistance of human colorectal cancer stem cells via PI3 K/Akt/NF-κB signaling pathway.Methods Suspended tumor cells were enriched from human colorectal cancer HCT-116 cells which cultured with serum-free medium,and used for cell expansion.HCT-116/oxaliplatin(L-OHP)resistant cells were cultured in the medium containing oxaliplatin.The effect of Zuojin Pill combined with L-OHP on chemotherapy tolerance was detected by cell counting kit-8(CCK-8).The expressions of CD133,Nanog,Oct4,CD44 and SOX2 in tumor stem cells after treatment with Zuojin Pill were detected by real-time PCR.The effects of Zuojin Pill on the protein expressions of AKT,p-AKT(Thr308 and Ser473),NF-κB and p-NF-κB in PI3 K/AKT/NF-κB signaling pathway in stem cells and drug resistant cells were detected by Western blot.Results Zuojin Pill could increase the sensitivity of HCT-116/SP cells to chemotherapeutics,in a dose-dependent manner.Zuojin Pill could significantly inhibit the mRNA expressions of CD44 and SOX2 in HCT-116/SP cells,and reduce the expressions of p-AKT and p-IκB in HCT-116/SP and HCT-116/L-OHP cells(P<0.01),in a dose-dependent manner.Conclusion Zuojin Pill can reverse the drug resistance of HCT-116/SP cells by decreasing the phosphorylation levels of p-AKT and p-IκB,inhibiting the activation of PI3 K/AKT/NF-κB signaling pathway,and reducing the expressions of stem cell marker CD44 and SOX2.

关 键 词：左金丸 结肠癌 肿瘤耐药 干细胞 PI3K/AKT NF-ΚB 

分 类 号：R285.5[医药卫生—中药学]