DUSP10通过增强肝癌细胞干性促进对仑伐替尼耐药的实验研究*

doi:10.3969/j.issn.1672-5069.2025.04.004

摘要/Abstract

摘要： 目的探讨双特异性蛋白磷酸酶10(DUSP10)通过调控肝癌细胞干性特征介导对仑伐替尼耐药的作用机制。方法构建Huh7-Resistant和Hep3B-Resistant仑伐替尼耐药细胞系,建立稳定过表达DUSP10的Huh7和PLC/PRF/5细胞及敲减DUSP10的Huh7-Resistant、Hep3B-Resistant和Hep-12细胞模型。采用Western blot法检测干性标志物(Nanog、BMI1和ABCG2)表达,采用CCK8法测定IC50值并计算耐药指数(RI)。结果耐药细胞系DUSP10表达量较野生型上调2.1～3.8倍(P<0.01);DUSP10过表达使Huh7和PLC/PRF/5细胞对仑伐替尼IC50分别从1.376 μM升至28.44 μM(RI=20.67)和4.118 μM升至18.01 μM(RI=4.37),干性基因表达上调1.5～2.3倍;敲减DUSP10使Huh7-Resistant、Hep3B-Resistant和Hep-12细胞的IC50分别降低6.53倍、12.02倍和3.29倍(均P<0.001),干性基因表达下调约40%～60%。结论 DUSP10通过上调Nanog/BMI1/ABCG2等干性基因表达显著降低肝癌细胞对仑伐替尼的敏感性,靶向干预DUSP10-干性通路可能逆转癌细胞耐药。

关键词: 肝细胞癌细胞, 双特异性蛋白磷酸酶10, 肝癌干细胞, 干性, 仑伐替尼, 耐药, 体外

Abstract: Objective This experiment aimed to investigate the mechanism by which dual specificity protein phosphatases 10(DUSP10) mediates lenvatinib resistance by through regulating stemness characteristics in hepatocellular carcinoma (HCC) in vitro. Methods Lenvatinib-resistant cell lines, e.g., Huh7-resistant and Hep3B-resistant, were established, and stable DUSP10-overexpressing (Huh7 and PLC/PRF/5) and knockdown (Huh7-resistant, Hep3B-resistant, Hep-12) cell models were constructed. Western blot was conducted to detect stemness markers (Nanog, BMI1, ABCG2) expression, and CCK-8 assay was performed to determine IC50 values and calculate the resistance index (RI). Results DUSP10 expression in resistant cell lines was up-regulated by 2.1 to 3.8 fold compared to in wild-type cells (P<0.01); overexpression of DUSP10 increased the IC50 of lenvatinib in Huh7 cells from 1.376 μM to 28.44 μM (RI=20.67) and in PLC/PRF/5 cells from 4.118 μM to 18.01 μM (RI=4.37), accompanied by a 1.5 to 2.3 fold up-regulation of stemness genes; conversely, DUSP10 knockdown reduced the IC50 in Huh7-resistant, Hep3B-resistant, and Hep-12 cells by 6.53 fold, 12.02 fold, and 3.29 fold, respectively (all P<0.001), with a 40% to 60% down-regulation of stemness genes. Conclusion DUSP10 significantly decreases the sensitivity of HCC cells to lenvatinib by probably up-regulating stemness-related genes, such as Nanog/BMI1/ABCG2, and targeting the DUSP10-stemness pathway might reverse drug resistance.

Key words: Hepatocellular carcinoma, Cancer stem cells, Dual specificity protein phosphatases 10, Stemness, Lenvatinib, Drug resistance, In vitro

李昂, 杨晓丹. DUSP10通过增强肝癌细胞干性促进对仑伐替尼耐药的实验研究^*[J]. 实用肝脏病杂志, 2025, 28(4): 493-496.

Li Ang, Yang Xiaodan. Mechanistic of DUSP10-mediated lenvatinib resistance in hepatocellular carcinoma by through cancerous stem cell regulation[J]. Journal of Practical Hepatology, 2025, 28(4): 493-496.

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DUSP10通过增强肝癌细胞干性促进对仑伐替尼耐药的实验研究^*

Mechanistic of DUSP10-mediated lenvatinib resistance in hepatocellular carcinoma by through cancerous stem cell regulation

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