| 摘要: |
| 目的 探讨三氧化二砷(ATO)在肝癌中对肽基脯氨酰基顺反异构酶 NIMA 相互作用蛋白 1(Pin1)表达的抑制作用及其分子机制。方法 使用 DepMap 数据库和 GEPIA 数据库中的数据分析 Pin1 在人肝癌细胞系和肝癌组织中的表达情况。以人肝癌细胞系 Huh7 和小鼠肝癌细胞系 H22 为细胞模型,通过 ATP 法检测 ATO 对肿瘤细胞活力的影响;通过蛋白质印迹法、免疫荧光染色和 qPCR 检测 ATO 对 Pin1 在蛋白质水平和转录水平表达的作用。用氯喹预处理 Huh7 细胞抑制溶酶体途径后,通过蛋白质印迹法和免疫荧光染色检测 ATO 对 Pin1 表达的调控作用。通过皮下荷瘤小鼠模型和免疫组织化学染色验证 ATO 在体内对肝癌细胞生长和 Pin1 表达的影响。采用 RNA 测序分析 ATO 可能影响的信号通路,并通过蛋白质印迹法和 qPCR 验证。结果 在 DepMap 数据库 23 种人肝癌细胞系和GEPIA 数据库人肝癌组织中, Pin1 的表达均呈现较高水平。在体外实验中, ATO 处理后 Huh7 和 H22 细胞的细胞活力均降低,细胞中 Pin1 在蛋白质和转录水平的表达均降低,但用氯喹抑制溶酶体途径逆转了 ATO 对 Pin1 表达的影响。在皮下荷瘤小鼠模型中, ATO 表现出一定的抗肿瘤效果,免疫组织化学染色显示 ATO 处理后 Pin1 表达和肿瘤细胞增殖受到抑制。在 ATO 处理的 H22 细胞中 Wnt/β- 连环素通路相关基因富集,抑制 H22 细胞中 Pin1 的表达后 β- 连环素表达减少。结论 ATO 通过溶酶体途径抑制 Pin1 表达,以及影响 Wnt/β- 连环素信号通路来抑制肝癌细胞增殖。 |
| 关键词: 三氧化二砷 肽基脯氨酰基顺反异构酶NIMA相互作用蛋白1 肝癌 Wnt/β-连环素 |
| DOI：10.16781/j.CN31-2187/R.20230592 |
| 投稿时间：2023-10-30修订日期：2024-01-18 |
| 基金项目:上海市卫生健康委员会卫生行业临床研究专项(202340091). |
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| Arsenic trioxide inhibits proliferation of hepatocellular carcinoma cells by decreasing expression of Pin1 and regulating Wnt/β-catenin pathway |
| LI Xin,WAN Qianqian,HUANG Nian*,WAN Xuying* |
(Department of Integrative Medicine, The Third Affiliated Hospital of Naval Medical University(Second Military Medical University), Shanghai 200438, China
*Corresponding authors) |
| Abstract: |
| Objective To investigate the inhibitory effect of arsenic trioxide (ATO) on the expression of peptidylprolyl cis-trans isomerase NIMA-interacting 1 (Pin1) in liver cancer and its molecular mechanism. Methods The expression of Pin1 in human hepatocellular carcinoma cell lines and liver cancer tissue was analyzed using DepMap database and GEPIA database. Human hepatocellular carcinoma cell line Huh7 and mouse hepatocellular carcinoma cell line H22 were used as cell models to detect the effect of ATO on tumor cell viability by adenosine triphosphate method. The effects of ATO on Pin1 expression at protein and transcription levels were detected by Western blotting, immunofluorescence staining and quantitative polymerase chain reaction (qPCR). After pretreatment with chloroquine to inhibit lysosome pathway in Huh7 cells, the regulation effect of ATO on Pin1 expression was detected by Western blotting and immunofluorescence staining. The effects of ATO on hepatoma cell growth and Pin1 expression in vivo were verified by subcutaneous tumor-bearing mouse model and immunohistochemical staining. The possible signaling pathways affected by ATO were analyzed by RNA sequencing and were verified by Western blotting and qPCR. Results The expression of Pin1 in 23 kinds of human hepatocellular carcinoma cell lines in DepMap database and human hepatocellular carcinoma tissue in GEPIA database showed a high level. In vitro, the viability of Huh7 and H22 cells was decreased after ATO treatment, and the protein and transcription levels of Pin1 were decreased. The effect of ATO on Pin1 expression was reversed after inhibiting lysosomal pathway by chloroquine. In the subcutaneous tumor-bearing mouse model, ATO showed certain anti-tumor effects, and immunohistochemical staining showed that Pin1 expression and tumor cell proliferation were inhibited after ATO treatment. The genes related to Wnt/β-catenin pathway were enriched in ATO-treated H22 cells, and the expression of β-catenin was decreased after inhibiting Pin1 expression in H22 cells. Conclusion ATO inhibits proliferation of hepatocellular carcinoma cells by inhibiting Pin1 expression through lysosomal pathway and affecting Wnt/β-catenin signaling pathway. |
| Key words: arsenic trioxide peptidyl-prolyl cis-trans isomerase NIMA-interacting 1 liver cancer Wnt/β-catenin |
