葛根素与尿路上皮癌胚抗原1表达在食管癌细胞凋亡和自噬中的关系  

Relationship between puerarin and urothelial carcinoembryonic antigen 1 expression in apoptosis and autophagy of esophageal cancer cells

作　　者：李晶 王菊美[1] 田敬荣 毕丽华 郜园园 尚红 LI Jing;WANG Jumei;TIAN Jingrong;BI Lihua;GAO Yuanyuan;SHANG Hong(The Fifth Department of Oncology,Handan Central Hospital,Handan,Hebei 056000,China)

机构地区：[1]邯郸市中心医院肿瘤内五科,河北邯郸056000

出　　处：《中国临床研究》2025年第1期82-86,共5页Chinese Journal of Clinical Research

基　　金：河北省卫生健康委员会科研项目(20220316);邯郸市科学技术研究与发展计划项目(22422083023ZC)。

摘　　要：目的 探讨葛根素影响食管癌细胞凋亡和自噬的分子机制。方法 使用浓度为0、200、500、800μmol/L的葛根素处理人食管癌细胞EC109,CCK-8法检测细胞增殖活性;划痕试验检测细胞迁移能力;Transwell试验检测细胞侵袭能力;Western blot法检测细胞凋亡相关蛋白(促凋亡蛋白Bax和Bad、抗凋亡蛋白Bcl-2)及自噬相关蛋白p62、Beclin-1和ATG5的表达水平;实时荧光定量聚合酶链反应检测尿路上皮癌胚抗原1(UCA1)表达水平。通过慢病毒转染细胞的方法构建稳定过表达UCA1的EC109细胞株,分为稳定过表达UCA1的细胞(UCA1-OE)及阴性对照细胞(UCA1-OENC),以相同500μmol/L葛根素对两组细胞进行处理,细胞免疫荧光试验检测两组细胞凋亡和自噬蛋白表达水平。结果 200、500、800μmol/L葛根素处理显著抑制EC109细胞的增殖、迁移和侵袭活性(P<0.01),促进Bax、Bad、p62、Beclin-1、ATG5蛋白表达(P<0.01),抑制Bcl-2蛋白表达(P<0.01),且均呈浓度依赖性;与0μmol/L(1.833±0.061)相比,200、500、800μmol/L葛根素处理显著抑制UCA1蛋白表达(UCA1相对表达水平分别为1.490±0.056、1.120±0.044和0.783±0.015,F=274.876,P<0.01),也呈浓度依赖性。500μmol/L葛根素处理后,与UCA1-OENC细胞相比,过表达UCA1的UCA1-OE细胞呈现更低的Bax、Bad、p62、Beclin-1、ATG5表达水平,更高的Bcl-2表达水平(P<0.05)。结论 葛根素通过抑制UCA1的表达诱导食管癌细胞凋亡和自噬。Objective To explore the molecular mechanism of puerarin on apoptosis and autophagy in esophageal cancer cells.Methods Human esophageal cancer EC109 cells were treated with puerarin at 0,200,500,800μmol/L.The cell proliferation,migration and invasion ability were measured by CCK-8 method,the scratch assay and transwell test,respectively.The expression levels of apoptosis-related proteins(pro-apoptotic proteins Bax and Bad,anti-apoptotic protein Bcl-2)and autophagy-related proteins p62,Beclin-1 and ATG5 were detected by Western blot.The expression level of urothelial carcinoembryonic antigen 1(UCA1)was detected by real-time fluorescence quantitative polymerase chain reaction.EC109 cell lines stably overexpressing UCA1 were constructed by lentiviral transfection and divided into stable overexpressing UCA1 cells(UCA1-OE group)and negative control cells(UCA1-OENC group).The two groups of cells were treated with the same 500μmol/L puerarin,and the expression levels of cells apoptosis and autophagy protein were detected using cellular immunofluorescence assay.Results Puerarin treatment with 200,500 and 800μmol/L significantly inhibited the proliferation,migration and invasion activities of EC109 cells(P<0.01),promoted the expression of Bax,Bad,p62,Beclin-1 and ATG5 proteins(P<0.01)and inhibited the expression of Bcl-2 protein(P<0.01),all of which were concentration-dependent.Compared with 0μmol/L(1.833±0.061),puerarin treatments of 200,500 and 800μmol/L significantly inhibited the expression of UCA1 protein(the relative expression levels of UCA1 were 1.490±0.056,1.120±0.044 and 0.783±0.015,respectively,F=274.876,P<0.01),which was also concentration-dependent.After 500μmol/L puerarinin treatment,UCA1-OE cells overexpressing UCA1 showed lower expression levels of Bax,Bad,p62,Beclin-1 and ATG5,as well as higher expression levels of Bcl-2 compared to UCA1-OENC cells(P<0.05).Conclusion Puerarin induces apoptosis and autophagy in esophageal cancer cells by inhibiting the expression of UCA1.

关 键 词：葛根素 尿路上皮癌胚抗原1 食管癌 凋亡 自噬 

分 类 号：R735.1[医药卫生—肿瘤]