| 连翘脂素通过抑制Wnt/β-catenin信号通路诱导结肠癌HCT116细胞凋亡和焦亡的机制 |
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| DOI： |
| 中文关键词: 结肠癌 连翘脂素 细胞凋亡 细胞焦亡 Wnt/β-catenin信号通路 |
| 英文关键词: Colon cancer Phillygenin Apoptosis Pyroptosis Wnt/β-catenin signaling pathway |
| 基金项目:1.江西省中医药管理局科技计划(编号:2023B0863)；2.江西省卫生健康委科技计划(编号:202410270)； |
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| 中文摘要: |
| 【】目的 探讨连翘脂素通过抑制Wnt/β-catenin信号通路诱导结肠癌HCT116细胞凋亡和焦亡的机制。方法? 在体外实验中,选用人结肠癌细胞HCT116进行实验,运用CCK-8法、流式细胞仪检测连翘脂素对HCT116细胞存活率、凋亡率并筛选连翘脂素的最适浓度,观察细胞增殖、侵袭、迁移、凋亡、焦亡等表型变化,采用JC-1染色观察HCT116细胞线粒体膜电位变化,采用Western blotting法、ELISA法、qRT-PCR法检测连翘脂素对HCT116细胞凋亡和焦亡相关蛋白表达、炎性因子表达等指标的影响,并使用平板克隆实验、划痕实验、Transwell实验检测HCT116细胞增殖、侵袭、迁移能力。在体内实验中,构建结肠癌裸鼠成瘤模型,分为对照组和实验组各5例,对照组注射0.1% DMSO,实验组注射连翘脂素,观察裸鼠瘤体大小、体重、生存情况及瘤体组织相关蛋白表达和炎性因子活性。结果 连翘脂素对HCT116细胞的最适加药浓度为20mmol/L。连翘脂素显著抑制HCT116的细胞克隆形成率、细胞侵袭个数和划痕愈合率(IC50为10.75~18.80 mmol/L),并诱导细胞凋亡和焦亡,表现为Bax、cleaved Caspase-3、cleaved PARP、GSDME-N、cleaved IL-1β、IL-1β、IL-18、IL-1β mRNA和IL-18mRNA表达升高,Bcl-2、β-catenin、c-Myc、MMP2表达降低。结论 连翘脂素能够通过抑制Wnt/β-catenin信号通路诱导结肠癌HCT116细胞凋亡和焦亡,为连翘脂素类抗肿瘤药物的开发提供理论依据和实验支持。 |
| 英文摘要: |
| Objective? To investigate the mechanism by which Phillygenin induces apoptosis and pyroptosis in HCT116 colon cancer cells by inhibiting the Wnt/β-catenin signaling pathway. Method? In vitro experiments were conducted using human colon cancer cell line HCT116. The CCK-8 assay and flow cytometry were employed to determine the optimal concentration of Phillygenin by measuring the cell viability and apoptosis rate of HCT116 cells. Phenotypic changes in cell proliferation, invasion, migration, apoptosis, and pyroptosis were observed. JC-1 staining was used to monitor the mitochondrial membrane potential of HCT116 cells. Western blotting, ELISA, and qRT-PCR were utilized to assess the effects of Phillygenin on the expression of apoptosis- and pyroptosis-related proteins, and inflammatory factors. The proliferative, invasive, and migratory abilities of HCT116 cells were evaluated using colony formation assays, scratch assays, and Transwell assays. In vivo experiments involved the establishment of a colon cancer xenograft model in nude mice. The mice were divided into control and experimental groups (n=5 each). The control group was injected with 0.1% DMSO, while the experimental group was injected with Phillygenin. Tumor size, body weight, survival status, and the expression of related proteins and inflammatory factor activity in tumor tissues were monitored. Results? The optimal concentration of Phillygenin for HCT116 cells was 20 mmol/L. Phillygenin significantly inhibited the colony formation rate, number of invasive cells, and scratch wound healing rate of HCT116 cells (IC50: 10.75–18.80 mmol/L). It also induced apoptosis and pyroptosis, as evidenced by increased expression of Bax, cleaved Caspase-3, cleaved PARP, GSDME-N, cleaved IL-1β, IL-1β, IL-18, IL-1β mRNA, and IL-18 mRNA, and decreased expression of Bcl-2, β-catenin, c-Myc, and MMP2. Conclusion? Phillygenin can induce apoptosis and pyroptosis in HCT116 colon cancer cells by inhibiting the Wnt/β-catenin signaling pathway, providing theoretical basis and experimental support for the development of Phillygenin-based anti-tumor drugs. |
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