Smac多肽对食管鳞状细胞癌耐药性调节机制的研究  

Research on the regulatory mechanism of Smac peptide on drug resistance in esophageal squamous cell carcinoma

作　　者：杨勇伟[1] 艾力江·多力坤 金澄宇[1] YANG Yong-wei;AILIJIANG Duolikun;JIN Cheng-yu(Department of Thoracic Surgery,People′s Hospital of Xinjiang Uygur Autonomous Region,Uygur Autonomous Region 830000,Xinjiang,China)

机构地区：[1]新疆维吾尔自治区人民医院胸外科,新疆乌鲁木齐830000

出　　处：《广东医学》2024年第4期425-432,共8页Guangdong Medical Journal

基　　金：新疆维吾尔自治区自然科学基金(2020D01C119)。

摘　　要：目的探究Smac多肽(Smac-N7)对食管癌鳞状细胞系Eca-109和紫杉醇(PTX)耐药株(Eca-109/PTX)耐药性的影响和作用机制。方法采用小剂量间歇诱导法建立Eca-109/PTX细胞株,CCK-8法检测PTX对Eca-109和Eca-109/PTX的半数抑制浓度(IC_(50))。使用PTX和Smac-N7分别或联合处理Eca-109或Eca-109/PTX细胞,并将细胞分为:Eca-109组、Eca-109+PTX组、Eca-109+Smac-N7组、Eca-109+PTX+Smac-N7组;Eca-109/PTX组、Eca-109/PTX+PTX组、Eca-109/PTX+Smac-N7组、Eca-109/PTX+PTX+Smac-N7组。采用流式细胞术、试剂盒和Western blot法分别检测各组Eca-109和Eca-109/PTX细胞的凋亡率和细胞中caspase-3活性与Bcl-2、Bax蛋白的表达水平。结果CCK-8法检测结果显示,随浓度的增加,PTX对Eca-109与Eca-109/PTX的生长抑制率均明显增加(P<0.05),且PTX对Eca-109的IC_(50)[(0.08±0.01)μg/mL]显著低于其对Eca-109/PTX的IC_(50)[(2.47±0.11μg/mL)](P<0.01)。与Eca-109组或Eca-109/PTX组相比,Eca-109+PTX组、Eca-109+PTX+Smac-N7组和Eca-109/PTX+Smac-N7组、Eca-109/PTX+PTX+Smac-N7组细胞中Bcl-2蛋白表达水平显著降低(P<0.05),但细胞凋亡率、细胞中caspase-3活性和Bax蛋白表达水平明显升高(P<0.05)。与Eca-109+PTX组或Eca-109/PTX+PTX组相比,Eca-109+PTX+Smac-N7组和Eca-109/PTX+PTX+Smac-N7组细胞中Bcl-2蛋白表达水平明显降低(P<0.05),而细胞凋亡率、细胞中caspase-3活性和Bax蛋白表达水平明显升高(P<0.05)。结论Smac-N7可在体外通过促进细胞凋亡来改善Eca-109细胞及其耐药株对PTX的敏感度。Objective To investigate the effects and the underlying mechanisms of Smac peptide(Smac-N7)on the resistance of esophageal squamous cell carcinoma cell line Eca-109 and paclitaxel(PTX)-resistant strain(Eca-109/PTX).Methods The Eca-109/PTX cell line was established using a low-dose intermittent induction method.The half-maximal inhibitory concentration(IC_(50))of PTX on Eca-109 and Eca-109/PTX was detected using the CCK-8 assay.Eca-109 and Eca-109/PTX cells were treated with PTX and Smac-N7 alone or in combination and were divided into groups:Eca-109 group,Eca-109+PTX group,Eca-109+Smac-N7 group,and Eca-109+PTX+Smac-N7 group;Eca-109/PTX group,Eca-109/PTX+PTX group,Eca-109/PTX+Smac-N7 group,and Eca-109/PTX+PTX+Smac-N7 group.Flow cytometry,assay kits,and Western blot were used to detect apoptosis rates and the expression levels of caspase-3,Bcl-2,and Bax proteins in Eca-109 and Eca-109/PTX cells.Results The CCK-8 assay results showed that with increasing concentrations,the growth inhibition rates of PTX on Eca-109 and Eca-109/PTX increased significantly(P<0.05),and the IC_(50) of PTX on Eca-109[(0.08±0.01)μg/mL]was significantly lower than that on Eca-109/PTX[(2.47±0.11)μg/mL](P<0.01).Compared with the Eca-109 group or Eca-109/PTX group,the expression levels of Bcl-2 protein in Eca-109+PTX group,Eca-109+PTX+Smac-N7 group,Eca-109/PTX+Smac-N7 group,and Eca-109/PTX+PTX+Smac-N7 group cells decreased significantly(P<0.05),while the apoptosis rate,caspase-3 activity,and Bax protein expression levels increased significantly(P<0.05).Compared with the Eca-109+PTX group or Eca-109/PTX+PTX group,the expression levels of Bcl-2 protein in Eca-109+PTX+Smac-N7 group and Eca-109/PTX+PTX+Smac-N7 group cells decreased significantly(P<0.05),while the apoptosis rate,caspase-3 activity,and Bax protein expression levels increased significantly(P<0.05).Conclusion Smac-N7 can improve the sensitivity of Eca-109 cells and its PTX-resistant strain to PTX in vitro by promoting cell apoptosis.

关 键 词：食管鳞状细胞癌 Smac多肽 耐药性 

分 类 号：R735.1[医药卫生—肿瘤] R329.2[医药卫生—临床医学]