鼻咽清毒颗粒通过调节铜离子转运蛋白CTR1逆转鼻咽癌细胞顺铂耐药的作用效应及机制研究  

Effect and mechanism of Biyan Qingdu Granules on reversing cisplatin resistance in nasopharyngeal carcinoma cells by regulating copper ion transporter CTR1

作　　者：石子叶 林娟[2] 杨冰 徐科一 张铁军[3,4] 李鹏 徐旭[3,4] SHI Ziye;LIN Juan;YANG Bing;XU Keyi;ZHANG Tiejun;LI Peng;XU Xu(Graduate School,Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China;Guangzhou Baiyunshan Hutchison Whampoa Chinese Medicine Co.,Ltd.,Guangzhou 510515,China;Tianjin Key Laboratory of Quality Marker of Traditional Chinese Medicine,Institute of Pharmaceutical Research,Tianjin 300301,China;National and Local Joint Engineering Laboratory of Modern Preparation and Quality Control Technology,Institute of Pharmaceutical Research,Tianjin 300301,China)

机构地区：[1]天津中医药大学研究生院,天津301617 [2]广州白云山和记黄埔中药有限公司,广东广州510515 [3]天津药物研究院天津市中药质量标志物重点实验室,天津300301 [4]天津药物研究院现代制剂与质量控制技术国家地方联合工程实验室,天津300301

出　　处：《现代药物与临床》2024年第8期1937-1943,共7页Drugs & Clinic

基　　金：广州市科技计划项目(20212210006)。

摘　　要：目的观察鼻咽清毒颗粒逆转鼻咽癌细胞顺铂耐药的作用效应,并探讨其作用机制。方法采用MTS法检测系列质量浓度鼻咽清毒颗粒对人鼻咽癌CNE-2、人鼻咽癌顺铂耐药CNE2/DDP细胞增殖的抑制作用;计算鼻咽清毒颗粒对CNE-2、CNE2/DDP细胞的耐顺铂半数抑制浓度(IC_(50))的影响;流式细胞技术测定CNE2/DDP的细胞凋亡情况;采用Western blotting法检测CNE2/DDP细胞中铜转运蛋白和抗凋亡蛋白的表达。结果CNE2/DDP细胞的顺铂耐药指数为RI值为3.03;与顺铂组比较,鼻咽清毒颗粒0.1、1、10 mg/mL组分别与系列质量浓度的顺铂联用,对CNE-2细胞顺铂IC50分别降低12.19%、46.37%、63.81%;CNE2/DDP细胞顺铂IC50分别降低61.75%、70.42%、83.82%。流式细胞技术检测结果表明,与顺铂组比,顺铂+鼻咽清毒颗粒0.1、1、10 mg/mL组CNE2-DDP晚期凋亡或坏死细胞增加(P<0.01);Western blotting法检测结果表明,与顺铂组比较,顺铂+鼻咽清毒颗粒0.1、1、10 mg/mL组铜特异性转运蛋白(CTR1)蛋白的表达均显著升高,B细胞淋巴瘤2(Bcl-2)、骨髓细胞白血病序列1(Mcl-1)的表达显著降低(P<0.05、0.01)。结论鼻咽清毒颗粒具有增强鼻咽癌细胞顺铂敏感性、逆转CNE2/DDP细胞顺铂耐药作用,其作用机制可能与增加CTR1蛋白表达,降低Bcl-2、Mcl-1蛋白表达有关。Objective To observe the effect of Biyan Qingdu Granules on reversing cisplatin resistance in nasopharyngeal carcinoma cells,and to explore its mechanism.Methods To detect the inhibitory effect of Biyan Qingdu Granules on the proliferation of CNE-2 and CNE2/DDP cells by MTS method.To calculate the effect of Biyan Qingdu Granules on cisplatin inhibiting IC_(50) in CNE-2 and CNE2/DDP cells.The apoptosis of CNE2/DDP cells was determined by flow cytometry.The expressions of copper transporter and anti-apoptotic protein in CNE2/DDP cells were detected by Western Blotting.Results The cisplatin resistance index of CNE2/DDP cells was 3.03.Compared with cisplatin,the IC50 of cisplatin in CNE-2 cells of 0.1,1 and 10 mg/mL of Biyan Qingdu Granules was decreased by 12.19%,46.37%and 63.81%,respectively.Cisplatin IC50 in CNE2/DDP cells decreased by 61.75%,70.42%and 83.82%,respectively.The results of flow cytometry showed that compared with cisplatin group,CNE2/DDP in Biyan Qingdu Granules 0.1,1,10 mg/mL groups increased late apoptosis or necrosis cells(P<0.01).Western Blotting test results showed that compared with cisplatin group,the expression of copper specific transport protein(CTR1)in cisplatin+Biyan Qingdu Granules 0.1,1 and 10 mg/mL groups was significantly increased.The expressions of B-cell lymphoma 2(Bcl-2)and myeloid leukemia sequence 1(Mcl-1)were significantly decreased(P<0.05,0.01).Conclusion Biyan Qingdu Granules can enhance the cisplatin sensitivity of nasopharyngeal carcinoma cells and reverse the cisplatin resistance of CNE2/DDP cells,and its mechanism may be related to increasing the expression of CTR1 protein and decreasing the expression of Bcl-2 and Mcl-1 protein.

关 键 词：鼻咽清毒颗粒 鼻咽癌 顺铂 耐药 铜特异性转运蛋白 B细胞淋巴瘤2 骨髓细胞白血病序列1 

分 类 号：R979.1[医药卫生—药品]