| HBV HBx 蛋白调控肝癌细胞增殖的机制研究 |
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| 引用本文： | 宋平辉,张毅,吴忠坤,李春博,李勤新. HBV HBx 蛋白调控肝癌细胞增殖的机制研究[J]. 医学分子生物学杂志, 2023, 20(1): 70-77. DOI: 10.3870/j.issn.1672-8009.2023.01.012 |
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| 作者姓名： | 宋平辉 张毅 吴忠坤 李春博 李勤新 |
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| 作者单位： | 1 陕西省核工业二一五医院普通外科 陕西省咸阳市, 712000 2 兴平市人民医院儿科 陕西省兴平市, 713199 |
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| 摘 要： | 目的 探讨 HBV HBx 蛋白调控肝癌细胞增殖的潜在机制。 方法 过表达 HBx 后, 检测肝癌细胞HepG2 的增殖水平。 在过表达 Flag-HBx 的肝癌细胞 HepG2 中免疫共沉淀 HBx 后进行蛋白质质谱鉴定,Score 阈值设定为大于 50, Coverage 阈值设定为大于 10。 过表达 HBx 的肝癌细胞 HepG2 中, 使用 siRNA 敲低质谱鉴定的相应基因, 检测 HepG2 的增殖水平。 结果 过表达 Flag-HBx 后, 肝癌细胞 HepG2 的增殖水平上升 (P< 0. 05)。 与对照组比较, 敲低 MCUR1 后肝癌细胞 HepG2 的增殖水平下降 (P< 0. 05)。 与对照组比较, 同时过表达 MCUR1 和 HBx 能够促进肝癌细胞 HepG2 的增殖 (P< 0. 05), 而仅过表达 MCUR1 对肝癌细胞 HepG2 的增殖无显著影响。 与过表达 HBx 组比较, 过表达 HBx 的同时敲低 MCUR1 时肝癌细胞HepG2 的增殖水平下降 (P< 0. 05)。 与对照组比较, 过表达 HBx 后肝癌细胞 HepG2 的 ROS 水平上升 (P<0. 05); 与对照组比较, 过表达 HBx 的同时敲低 MCUR1 时肝癌细胞 HepG2 的 ROS 水平无显著变化。 与对照组或过表达 HBx 组比较, 同时过表达 MCUR1 和 HBx 能够增加肝癌细胞 HepG2 的 ROS 水平 (P< 0. 05)。与过表达 HBx 组比较, 过表达 HBx 的同时敲低 MCUR1 时肝癌细胞 HepG2 的增殖水平下降 (P< 0. 05), 但这种下降能被 H2O2恢复。 与对照组比较, 过表达 HBx 后肝癌细胞 HepG2 中转录因子 Nrf2 在细胞核中的定位增加 (P< 0. 05), 而过表达 HBx 的同时敲低 MCRU1 后 Nrf2 在细胞核中的定位无显著变化。 与对照组比较, 过表达 HBx 后肝癌细胞 HepG2 中转录因子 Notch 的激活形式 NICD1 在细胞核中的定位增加 ( P<0. 05), 而过表达 HBx 的同时 Nrf2 抑制剂 ML385 处理后 NICD1 在细胞核中的定位无显著变化。 与过表达HBx 组比较, 过表达 HBx 的同时 ML385 处理后肝癌细胞 HepG2 的增殖水平下降 (P< 0. 05)。 与过表达HBx 组比较, 过表达 HBx 的同时 Notch 抑制剂 IMR-1 处理后肝癌细胞 HepG2 的增殖水平下降 (P< 0. 05)。 结论 HBx 促进 HepG2 细胞增殖与 MCUR1 / ROS / Nrf2 / Notch 轴有关。
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| 关 键 词： | HBx 肝癌 增殖 ROS MCUR1 |
Mechanism of HBV HBx Protein on Liver Cancer Cell ProliferationRegulation |
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| SONG Pinghui,ZHANG Yi,WU Zhongkun,LI Chunbo,LI Qinxin. Mechanism of HBV HBx Protein on Liver Cancer Cell ProliferationRegulation[J]. Journal of Medical Molecular Biology, 2023, 20(1): 70-77. DOI: 10.3870/j.issn.1672-8009.2023.01.012 |
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| Authors: | SONG Pinghui ZHANG Yi WU Zhongkun LI Chunbo LI Qinxin |
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| Affiliation: | 1 Department of General Surgery, the 215th Hospital of Shaanxi Provincial Nuclear Industry, Xianyang, Shaanxi, 712000, China 2 Department of Pediatrics, Xingping Municipal People’ s Hospital, Xingping, Shaanxi, 713199,China |
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| Abstract: | Objective To explore the potential mechanism of HBV HBx protein on liver cancercell proliferation regulation. Methods After HBx overexpression, the proliferation of HEPG2 livercancer cells was detected. HBx was immunomotized co-precipitated in the Flag-HBx overexpressedHEPG2 cells, and then protein mass spectrometry was performed, the SCORE threshold was set tobe greater than 50, and the Coverage threshold was set to be greater than 10. In HBx overexpressedHEPG2 cells, siRNAs were used to knock down the corresponding genes identified by the massspectrometry, and the proliferation of cells was detected. Results The proliferation level of HEPG2cells was increased after the expression of Flag-HBX, ( P < 0. 05). Compared with the controlgroup, the proliferation of HEPG2 cells was decreased after the knock-down of Mcur1 (P< 0. 05).Compared with the control group, the overexpression of Mcur1 and HBx at the same time can promote the proliferation of HEPG2 (P< 0. 05), and the overexpression of MCUR1 only has no effecton the proliferation of HEPG2. Compared with the HBx group, the proliferation of HEPG2 cells withoverexpressed HBx and lowly expressed MCUR1 was decreases (P< 0. 05). Compared with the control group, the ROS level of HEPG2 after HBx overexpression was increased (P < 0. 05), and ithad no significant change in HEPG2 with overexpression of HBx and knock-down of MCUR1 at thesame time. Compared with the control group and the HBx group, the overexpression of MCUR1 andHBx at the same time can increase the ROS level of HEPG2 (P < 0. 05). Compared with the HBxgroup, the proliferation of HEPG2 was decresed when HBx was overexpressed and the MCUR1 wasknock down (P< 0. 05), however, this decreasing could be restored by H2O2treatment. Comparedwith the control group, the positioning of nuclear transcription factor Nrf2 was increases in the HBxoverexpressed group (P< 0. 05), while it had no significant change in cells with HBx overexpression and MCUR1 knock-down at the same time. Compared with the control group, the activated form ofNOCD1 in the nucleus was increased, and it had no significant change after using ML385 in HBx overexpressed cells. Compared with the HBx group, the proliferation level of HEPG2, HEPG2 after ML385expressed HBX at the same time as HBX (P<0. 05). Compared with the HBX group, the proliferationof HEPG2 was decreased after treatment of NOTCH inhibitor IMR-1 or ML385. Conclusion HBx promotes HepG2 cell proliferation through the MCUR1/ ROS/ Nrf2/ Notch axis |
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| Keywords: | HBX liver cancer proliferation ROS MCUR1 |
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