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投稿时间：2024-11-06 网络发布日期：2025-01-20
投稿时间：2024-11-06 网络发布日期：2025-01-20
中文摘要: 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、800 μmol/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500 μmol/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、800 μmol/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、800 μmol/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。500 μmol/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。指标中的差异亦有统计学意义（P＜0.05）。不同年龄组患者部分指标的差异有统计学意义。结论 不同类型、不同性别错(牙合)畸形患者的下牙槽神经前环及其他颏孔区解剖存在差异。正(牙合)手术时应予以足够重视。
Abstract: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, 800 μmol/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 500 μmol/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 800 μmol/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 0 μmol/L (1.833±0.061), puerarin treatments of 200, 500 and 800 μmol/ 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 500 μmol/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.
文章编号： 中图分类号：R735.1 文献标志码：A
基金项目: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）
附件
| Author Name | Affiliation |
| LI Jing, WANG Jumei, TIAN Jingrong, BI Lihua, GAO Yuanyuan, SHANG Hong | The Fifth Department of Oncology, Handan Central Hospital, Handan, Hebei 056000, China |
引用文本：
李晶，王菊美，田敬荣，毕丽华，郜园园，尚红.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[J].中国临床研究,2025,38(1):82-86.
李晶，王菊美，田敬荣，毕丽华，郜园园，尚红.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[J].中国临床研究,2025,38(1):82-86.