ASPP2重组腺病毒通过调控NF-κB信号通路抑制DEN诱导的小鼠肝细胞癌发生*

doi:10.3969/j.issn.1672-5069.2024.02.003

实用肝脏病杂志 ›› 2024, Vol. 27 ›› Issue (2): 169-172, 173.doi: 10.3969/j.issn.1672-5069.2024.02.003

ASPP2重组腺病毒通过调控NF-κB信号通路抑制DEN诱导的小鼠肝细胞癌发生^*

高明慧, 柴梦音, 寇卜心, 豆双双, 刘晓霓

北京市首都医科大学附属北京佑安医院北京肝病研究所

收稿日期:2023-05-15 出版日期:2024-02-10 发布日期:2024-03-08
通讯作者: 刘晓霓,E-mail: liuxiaoni888@ccmu.edu.cn
作者简介:高明慧,女,23岁,硕士研究生。E-mail: minghui512484@163.com
共同第一作者:柴梦音,女,33岁,检验师。E-mail: mengyin618@126.com
基金资助:
^*北京市自然科学基金资助项目(编号:7192084);首都卫生发展科研专项基金资助项目(编号:2020-2-1152);北京市公共医学研究所发展改革试点研究项目(编号:京医研2021-10)

ASPP2 recombinant adenovirus inhibits DEN-induced hepatocarcinogenesis in mice by regulating NF-κB signaling pathway

Gao Minghui, Chai Mengyin, Kou Buxin, et al.

Beijing Institute of Liver Diseases, You 'an Hospital, Capital Medical University, Beijing 100069, China

Received:2023-05-15 Online:2024-02-10 Published:2024-03-08

摘要/Abstract

摘要： 目的研究ASPP2重组腺病毒(ASPP2-ad)对二乙基亚硝胺(DEN)诱导的小鼠肝细胞癌(HCC)发生的抑制作用及其可能的作用机制。方法采用DEN腹腔注射联合0.005% DEN饮水方法构建小鼠HCC模型。将动物分为DEN处理组和DEN-ASPP2-ad处理组, 每组10只。使用小动物超声成像系统动态观察小鼠HCC形成情况, 大体记数肝脏肿瘤数量, 采用免疫组化法检测肿瘤组织Ki67表达, 采用流式多重蛋白定量技术检测小鼠血清IL-1β、IL-6、KC、IL-2和TNFα水平, 采用Western blot法检测组织AFP、caspase3、cyclinD1、PCNA、p-IKK、IKK、p-IκB、IκB、p-p65和p65蛋白表达, 采用实时定量PCR法检测Nfatc1 mRNA水平。结果 DEN诱导24周后, DEN组小鼠肝脏肿瘤数为(9.9±1.9)个, 显著多于DEN联合ASPP2-ad组;DEN组小鼠Ki67阳性细胞数为(91.4±9.1)个, 显著多于DEN联合ASPP2-ad组;在实验40周末, DEN组小鼠生存率为65.2%, 显著低于DEN联合ASPP2-ad组的90.0%(P <0.05);DEN组小鼠血清ALT和AST水平分别为(271.5±143.8)U/L和(299.3±221.4)U/L, 均显著高于DEN-ASPP2-ad干预组;DEN联合ASPP2-ad组血清IL-6和TNFα水平分别为(8.1±1.6)MFI和(8.1±1.0)MFI, 均显著低于DEN组;DEN/ASPP2-ad处理组AFP、Cyclin D1和PCNA表达减弱, 而caspase-3表达增强, NF-κB信号通路蛋白(p-IKK、p-IκB和p-p65)表达减弱, p-IKK/IKKα、p-IκB/IκB和p-p65/p65比值也降低, NF-κB下游癌基因Nfatc1表达减弱(P<0.05)。结论 ASPP2-ad可能通过调控NF-κB通路显著抑制DEN诱导的炎性增殖反应, 阻抑HCC的发生, 值得深入研究。

关键词: 肝细胞癌, P53凋亡刺激蛋白2 , 二乙基亚硝胺, 核因子κB, 小鼠

Abstract: Objective The aim of this experiment was to investigate the inhibitory effect of apoptosis stimulating protein 2 of P53 (ASPP2) recombinant adenovirus (ASPP2-ad) on hepatocellular carcinogenesis induced by diethylnitrosamine (DEN) in mice. Methods The mouse liver cancer model was established by intraperitoneal injection of DEN at dose of 25 mg·kg^-1 and drinking water with 0.005% of DEN. The experiment was divided into two groups with 10 mice in each group, receiving DEN or DEN and ASPP2-ad combination intervention. The formation of liver cancer in mice was observed by ultrasonography. The number of liver tumors was recorded, and the expression of Ki67 was detected by immunohistochemistry. Serum IL-1β, IL-6, KC, IL-2 and TNFα levels were detected by flow multiprotein quantification technology. The protein expressions of AFP, caspase3, cyclin D1, PCNA, p-IKK, IKK, p-IκB, IκB, p-p65 and p65 were observed by Western blot, and the Nfatc1 mRNA level was assayed by real-time quantitative PCR. Results After 24 weeks of DEN induction, the number of liver tumors in DEN-intervened group was (9.9±1.9), significantly greater than in the DEN and ASPP2-ad combination intervened group; the number of Ki67 positive cells in DEN-intervened group was (91.4±9.1), significantly greater than in DEN/ASPP2-ad-intervened group; the survival rate in the DEN-intervened group was 65.2%, significantly lower than 90.0%(P <0.05)in DEN/ASPP2-ad-intervend group (P <0.05); serum ALT and AST levels were (271.5±143.8)U/L and (299.3±221.4)U/L, both significantly higher than in DEN/ASPP2-ad-intervend group; serum IL-6 and TNFα levels in DEN/ASPP2-ad-intervend group were (8.1±1.6)MFI and (8.1±1.0)MFI, both much lower than in DEN-intervened group; the cancerous AFP, Cyclin D1 and PCNA expression became weaker, the p-IKK, p-IκB and p-p65 expression down-regulated, the p-IKK/IKKα, p-IκB/IκB and p-p65/p65 ratio down-regulated, the Nfatc1 expression weaker, while the caspase-3 expression intensified (P<0.05) in DEN/ASPP2-ad-intervend group. Conclusion The ASPP2-ad could remarkably inhibit DEN-induced inflammatory proliferation reaction and the occurrence of liver cancer, which might be related to the regulation of NF-κB signal pathway, and warrants further investigation.

Key words: Hepatoma, p53 apoptosis-stimulating protein 2, Diethylnitrosamine , Nuclear factor κB, Mice

高明慧, 柴梦音, 寇卜心, 豆双双, 刘晓霓. ASPP2重组腺病毒通过调控NF-κB信号通路抑制DEN诱导的小鼠肝细胞癌发生^*[J]. 实用肝脏病杂志, 2024, 27(2): 169-172, 173.

Gao Minghui, Chai Mengyin, Kou Buxin, et al.. ASPP2 recombinant adenovirus inhibits DEN-induced hepatocarcinogenesis in mice by regulating NF-κB signaling pathway[J]. Journal of Practical Hepatology, 2024, 27(2): 169-172, 173.

参考文献

[1] Stauffer J K, Scarzello A J, Jiang Q, et al. Chronic inflammation, immune escape, and oncogenesis in the liver: a unique neighborhood for novel intersections. Hepatology, 2012, 56(4):1567-1574.
[2] He Q, Yang J, Jin Y. Development and validation of TACE refractoriness-related diagnostic and prognostic scores and characterization of tumor microenvironment infiltration in hepatocellular carcinoma. Front Immunol, 2022, 13:869993.
[3] Craig AJ, von Felden J, Garcia-Lezana T,et al. Tumour evolution in hepatocellular carcinoma. Nat Rev Gastroenterol Hepatol, 2020, 17(3):139-152.
[4] Majumder S, Roy S, Kaffenberger T, et al. Loss of metallothionein predisposes mice to diethylnitrosamine-induced hepatocarcinogenesis by activating NF-kappaB target genes. Cancer Res, 2010, 70(24):10265-10276.
[5] Yunfei Huo, Ke Cao, Buxin Kou, et al. TP53BP2: roles in suppressing tumorigenesis and therapeutic opportunities. Genes & Diseases, https://doi.org/10.1016/j.gendis.
[6] Liang B, Jiang Y, Song S, et al. ASPP2 suppresses tumour growth and stemness characteristics in HCC by inhibiting Warburg effect via WNT/β-catenin/HK2 axis. J Cell Mol Med, 2023, 27(5): 659-671.
[7] Wang S, Kou B, Chai M, et al. Knockout of ASPP2 promotes DEN-induced hepatocarcinogenesis via the NF-κB pathway in mice. Cancer Gene Ther, 2022, 29(2): 202-214.
[8] Chen R, Wang H, Liang B, et al.Downregulation of ASPP2 improves hepatocellular carcinoma cells survival via promoting BECN1-dependent autophagy initiation. Cell Death Dis, 2016, 7(12):e2512.
[9] Patel KD, Barasiya YV, Patel JB, et al. Apoptosis stimulating protein of p53 (ASPP) 1 and ASPP2 mRNA expression in oral cancer. Arch Oral Biol, 2020, 119:104920.
[10] Jiang F, Bian G, Li J.ASPP2 promotes cell apoptosis in cervical cancer through inhibiting autophagy. Exp Ther Med, 2022, 24(6): 726.
[11] Liu D, Ertay A, Hill C, et al. ASPP1 deficiency promotes epithelial-mesenchymal transition, invasion and metastasis in colorectal cancer. Cell Death Dis, 2020, 11(4):224.
[12] Wang CH, Li QY, Nie L,et al. LncRNA ANRIL promotes cell proliferation, migration and invasion during acute myeloid leukemia pathogenesis via negatively regulating miR-34a. Int J Biochem Cell Biol, 2020, 119:105666.
[13] Mori T, Okamoto H, Takahashi N, et al. Aberrant overexpression of 53BP2 mRNA in lung cancer cell lines. FEBS Lett, 2000, 465(2-3):124-128.
[14] Wu F, Zhang Y, Fang Y, et al. Elevated expression of inhibitor of apoptosis-stimulating protein of p53 (iASPP) and methyltransferase-like 3 (METTL3) correlate with poor prognosis in FIGO Ib1-IIa squamous cell cervical cancer. J Cancer, 2020, 11(9):2382-2389.
[15] Li S, Shi G, Yuan H, et al. Abnormal expression pattern of the ASPP family of proteins in human non-small cell lung cancer and regulatory functions on apoptosis through p53 by iASPP.Oncol Rep, 2012, 28(1):133-140.
[16] Lossos I S, Natkunam Y, Levy R, et al.Apoptosis stimulating protein of p53 (ASPP2) expression differs in diffuse large B-cell and follicular center lymphoma: correlation with clinical outcome. Leuk Lymphoma, 2002, 43(12): 2309-2317.
[17] 寇卜心, 柴梦音, 豆双双等. p53凋亡刺激蛋白2基因肝脏条件性敲除小鼠的构建和鉴定. 实用肝脏病杂志, 2022, 25(2): 170-173.
[18] Mak V C, Lee L, Siu M K, et al. Downregulation of ASPP2 in choriocarcinoma contributes to increased migratory potential through src signaling pathway activation. Carcinogenesis, 2013, 34(9): 2170-2177.
[19] Ju H, Lee K A, Yang M, et al. TP53BP2 locus is associated with gastric cancer susceptibility. Int J Cancer, 2005, 117(6): 957-960.
[20] Meng W D, Chu R X, Wang B Z, et al.Helicobacter pylori infection and expressions of apoptosis-related proteins p53, ASPP2 and iASPP in gastric cancer and precancerous lesions. Pathol Biol (Paris), 2013, 61(5):199-202.

ASPP2重组腺病毒通过调控NF-κB信号通路抑制DEN诱导的小鼠肝细胞癌发生^*

ASPP2 recombinant adenovirus inhibits DEN-induced hepatocarcinogenesis in mice by regulating NF-κB signaling pathway

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	哈鹏, 苏晓晨, 付宾鹏. 三期增强CT扫描联合血清AFP和VEGF水平诊断肝细胞癌患者临床研究^*[J]. 实用肝脏病杂志, 2024, 27(2): 251-254.
[2]	张亚龙, 胡冬梅, 赵晟, 孟海生. 超声引导下射频消融治疗小原发性肝癌疗效及死亡危险因素分析^*[J]. 实用肝脏病杂志, 2024, 27(2): 271-274.
[3]	崔甜甜, 张海波, 陈莉莉. 能谱CT增强扫描参数鉴别诊断肝血管瘤与肝细胞癌价值研究^*[J]. 实用肝脏病杂志, 2024, 27(2): 275-278.
[4]	魏野, 汪施妤, 葛高华, 井桂银. CT增强扫描与磁共振检查诊断肝血管瘤和肝细胞癌价值分析^*[J]. 实用肝脏病杂志, 2024, 27(2): 279-282.
[5]	何圣科, 蒲江汉, 冯海玲, 吴大平, 陈绵军, 陈辉. 肝细胞癌组织HOXC9和KIF4A表达及其临床意义探讨^*[J]. 实用肝脏病杂志, 2024, 27(1): 84-87.
[6]	郝永欣, 孟祥安, 骆允, 高美丽, 刘欢, 刘磊, 沈荣. 超声造影与增强CT检查诊断小肝癌效能比较^*[J]. 实用肝脏病杂志, 2024, 27(1): 88-91.
[7]	许梁, 梁宏伟, 杜圣, 黄爱娜, 张涛. 影响肝细胞癌患者肿瘤微血管侵犯的多因素分析^*[J]. 实用肝脏病杂志, 2024, 27(1): 92-95.
[8]	叶萌萌, 邓泽润, 张韬. 卡瑞利珠单克隆抗体联合阿帕替尼治疗中晚期肝细胞癌患者疗效及预后分析^*[J]. 实用肝脏病杂志, 2024, 27(1): 96-100.
[9]	仲金龙, 施琳. miR-556-3p通过调控PTEN/AKT信号通路影响肝细胞癌的转移活性^*[J]. 实用肝脏病杂志, 2023, 26(6): 781-784.
[10]	杨婉, 丁莉, 罗珂, 刘美. 肝细胞癌患者癌组织CK7和Glypican-3表达及其临床意义探讨^*[J]. 实用肝脏病杂志, 2023, 26(6): 883-886.
[11]	于双杰, 李元元, 毕京峰, 福军亮, 宓余强. ICIs联合TKIs治疗HBV相关肝细胞癌患者疗效研究^*[J]. 实用肝脏病杂志, 2023, 26(6): 891-894.
[12]	周莉, 丁云清, 袁凤英, 陈竹, 冯志伟. 超声造影判断乙型肝炎肝硬化患者肝内占位性病变性质价值研究^*[J]. 实用肝脏病杂志, 2023, 26(6): 903-906.
[13]	秦浩, 方春华, 张磊, 冉斌, 汪丽萍, 罗凌, 李应, 黄育红. 乙型肝炎病毒相关肝细胞癌癌组织ARHGAP18水平及其基因调控网络分析^*[J]. 实用肝脏病杂志, 2023, 26(5): 694-697.
[14]	武向慧, 李龙虎, 景斐华, 武鹏. 经导管动脉化疗栓塞联合射频消融治疗肝细胞癌患者疗效研究^*[J]. 实用肝脏病杂志, 2023, 26(5): 702-705.
[15]	刘清桂, 王紫君, 王敏君, 杜涵, 陈费. CDE诱导慢性肝损伤过程中肝细胞特异性过表达细胞周期抑制因子P21促进胆管上皮细胞转分化为成熟肝细胞研究^*[J]. 实用肝脏病杂志, 2023, 26(4): 466-471.