«上一篇/Previous Article|本期目录/Table of Contents|下一篇/Next Article»

《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:

30卷

期数:

2024年01期

页码:

11-14,34

栏目:

肝脏疾病专题

出版日期:

2024-01-01

文章信息/Info

Title:

The experimental study of miR-129-5p inhibiting proliferation， migration， and invasion of hepatocellular carcinoma by targeting to SALL4

文章编号:

1006-8147(2024)01-0011-05

作者:

张华¹; 2; 范松³; 刘冀琴²; 张学军¹

（1.天津医科大学基础医学院免疫学系，天津 300070；2.武警特色医学中心检验科，天津 300162；3.武警河北省总队医院检验科，石家庄 050000）

Author(s):

ZHANG Hua¹; 2; FAN Song³; LIU Jiqin²; ZHANG Xuejun¹

（1.Department of Immunology，School of Basic Medical Sciences，Tianjin Medical University，Tianjin 300070，China；2.Department of Clinical Laboratory，Characteristic Medical Center of Chinese People′s Armed Police Force，Tianjin 300162，China；3.Department of Clinical Laboratory，Armed Police Corps Hospital of Hebei Province，Shijiazhuang 050000，China）

关键词:

肝细胞癌; miR-129-5p; SALL4; 增殖; 侵袭; 迁移

Keywords:

hepatocellular carcinoma; miR-129-5p; SALL4; proliferation; invasion; migration

分类号:

R575.7

DOI:

10.20135/j.issn.1006-8147.2024.01.0011

文献标志码:

A

摘要:

目的：探讨miR-129-5p在肝细胞癌（HCC）发展中的作用及其机制。方法：通过实时荧光定量- PCR（qRT-PCR）检测miR-129-5p在正常血清及肝癌患者血清、人正常肝细胞 LO2 和人肝癌细胞系中的表达情况。采用脂质体转染技术将 miR-129-5p mimics 转染至人肝癌细胞HCCLM3 细胞，应用 Western 印迹检测人类婆罗双树样基因-4 （SALL4）和程序性死亡受体-配体1（PD-L1）蛋白的表达。通过 CCK-8 实验、克隆形成实验、细胞周期检测、细胞划痕实验和Transwell 实验等检测在体外过表达miR-129-5p对肝癌细胞增殖、迁移及侵袭能力的影响。此外，通过生物信息学工具、数据库分析和荧光素酶报告基因检测实验，探索miR-129-5p在HCC中的靶点，并探讨靶点SALL4是否介导miR-129-5p对HCC细胞的作用。结果：与正常血清相比，肝癌患者血清miRNA-129-5p表达量显著降低（t=13.32，P<0.001），与LO2相比，肝癌细胞系HepG2、Hep3B、HCCLM3、BEL-7402和QGY-7703的miR-129-5p表达量均显著降低（t=30.35、33.08、37.11、32.87、8.2，均P<0.05）。miR-129-5p过表达可以抑制肝癌细胞增殖、侵袭和转移。StarBase预测显示miR-129-5p与SALL4有潜在结合位点，双荧光素酶报告基因检测证明miR-129-5p与SALL4直接结合。与对照组相比，miR-129-5p表达后SALL4和PD-L1的蛋白水平明显降低（t=12.68、t=8.798，均P<0.05）。miR-129-5p可以通过调控SALL4的表达，抑制HCC细胞的活力、增殖、迁移和侵袭（F=26.11、147.2、4.302、321.3，均P<0.05）。结论：过表达miR-129-5p通过抑制SALL4表达，抑制肝癌细胞增殖、迁移和侵袭。

Abstract:

Objective： To explore the role of miR-129-5p in hepatocellular carcinoma（HCC） development and its mechanism. Methods： The expression of miR-129-5p in normal serum and serum of hepatocellular carcinoma patients， human normal hepatocyte LO2 and human hepatocellular carcinoma cell line was detected by quantitative real-time PCR（qRT-PCR）. Human hepatocellular carcinoma cells HCCLM3 were selected as the research objects， and miR-129-5p mimics was transfected into HCCLM3 cells by liposome transfection technology. Western blotting was used to detect the protein expression of Sal-like gene 4（SALL4） and programmed death receptor-ligand 1（PD-L1）. CCK-8 assay， colony formation assay， cell cycle detection， cell scratch assay and Transwell assay were used to detect the effects of miR-129-5p overexpression on the proliferation， migration and invasion of HCC in vitro. In addition， bioinformatics tools， database analysis， luciferase reporter gene detection and rescue experiments were used to explore the target of miR-129-5p in HCC and to explore whether the target SALL4 mediated the effect of miR-129-5p on HCC cells. Results： The expression of miR-129-5p was signifi-cantly decreased in the serum of HCC patients compared with normal serum（t=13.32，P<0.001）. Compared with LO2， the expression of miR-129-5p in liver cancer cell lines including HepG2， Hep3B， HCCLM3， BEL-7402 and QGY-7703 was significantly decreased（t=30.35， 33.08， 37.11， 32.87， 8.2， all P<0.05）. Overexpression of miR-129-5p inhibited the proliferation， invasion and metastasis of hepatocellular carcinoma cells. StarBase predicted that miR-129-5p had potential binding sites with SALL4， and dual luciferase reporter gene assay confirmed that miR-129-5p directly bound to SALL4. Compared with the control group， the protein levels of SALL4 and PD-L1 were significantly decreased after miR-129-5p expression（t=12.68， 8.798， both P<0.05）. miR-129-5p could inhibit the viability， proliferation， migration and invasion of HCC cells by regulating the expression of SALL4（F=26.11，147.2，4.302，321.3，all P<0.05）. Conclusion： Overexpression of miR-129-5p inhibits the proliferation， migration， and invasion of hepatocellular carcinoma by inhibiting SALL4 expression.

参考文献/References:

[1] PI?譙ERO F，DIRCHWOLF M，PESS A M G. Biomarkers in hepatocellular carcinoma: diagnosis, prognosis and treatment response assessment[J]. Cells，2020，9（6）：1370.
[2] LEE S K，LEE S W，JANG J W，et al. Immunological markers, prognostic factors and challenges following curative treatments for hepatocellular carcinoma[J]. Int J Mol Sci，2021，22（19）： 10271.
[3] CHEN Z，XIE H，HU M，et al. Recent progress in treatment of hepatocellular carcinoma[J]. Am J Cancer Res，2020，10（9）： 2993-3036.
[4] CUCARULL B，TUTUSAUS A. Hepatocellular carcinoma: molecular pathogenesis and therapeutic advances[J]. Cancer，2022，14（3）：621.
[5] KHARE S，KHARE T，RAMANATHAN R. Hepatocellular carcinoma: the role of microRNAs[J]. Biomolecules，2022，12（5）：645.
[6] ZENG A，YIN J，LI Y，et al. miR-129-5p targets Wnt5a to block PKC/ERK/NF-κB and JNK pathways in glioblastoma[J]. Cell Death Dis，2018，9（3）：394.
[7] GAO G，XIU D，YANG B，et al. MiR-129-5p inhibits prostate cancer proliferation via targeting ETV1[J]. Onco Targets Ther，2019， 12：3531-3544.
[8] XU C，DU Z，REN S，et al. MiR-129-5p sensitization of lung cancer cells to etoposide-induced apoptosis by reducing YWHAB[J]. J Ca-ncer， 2020，11（4）：858-866.
[9] HUGE N，REINKENS T，BUURMAN R，et al. MiR-129-5p exerts Wnt signaling-dependent tumor-suppressive functions in hepatocellular carcinoma by directly targeting hepatoma-derived growth factor HDGF[J]. Cancer Cell Int，2022，22（1）：192.
[10] GAO B，WANG L，ZHANG N，et al. miR-129-5p inhibits clear cell renal cell carcinoma cell proliferation, migration and invasion by targeting SPN[J]. Cancer Cell Int，2021，21（1）：263.
[11] FU X，YANG Y，ZHANG D. Molecular mechanism of albumin in suppressing invasion and metastasis of hepatocellular carcinoma [J].Liver Int，2022，42（3）：696-709.
[12] XU S，LI W，WU J，et al. The role of miR-129-5p in cancer: a novel therapeutic target[J]. Curr Mol Pharmacol，2022，15（4）： 647-657.
[13] LIU S，LIAO Q，XU W，et al. MiR-129-5p protects H9c2 cardiac myoblasts from hypoxia/reoxygenation injury by targeting TRPM7 and inhibiting NLRP3 inflammasome activation[J]. J Cardiovasc Pharmacol，2021，77（5）：586-593.
[14] HUGE N，REINKENS T，BUURMAN R，et al. MiR-129-5p exerts Wnt signaling-dependent tumor-suppressive functions in hepatocellular carcinoma by directly targeting hepatoma-derived growth factor HDGF[J]. Cancer Cell Int，2022，22（1）：192.
[15] SUN B，XU L，BI W，et al. SALL4 Oncogenic function in cancers: mechanisms and therapeutic relevance[J]. Int J Mol Sci，2022， 23（4）：2053.
[16] MOEIN S，TENEN D G，AMABILE G，et al. SALL4: an intriguing therapeutic target in cancer treatment[J]. Cells，2022，11（16）：2601.
[17] YIN F，HAN X，YAO S K，et al. Importance of SALL4 in the development and prognosis of hepatocellular carcinoma[J]. World J Gastroenterol， 2016，22（9）：2837-2843.
[18] SUN C，LAN P，HAN Q，et al. Oncofetal gene SALL4 reactivation by hepatitis B virus counteracts miR-200c in PD-L1-induced T cell exhaustion[J]. Nat Commun，2018，9(1）：1241.

相似文献/References:

[1]周冷潇,韩 涛.慢性乙型病毒性肝炎肝硬化发生肝细胞癌的危险因素分析[J].天津医科大学学报,2017,23(03):214.
　ZHOU Leng-xiao,HAN Tao.Risk factors of hepatocellular carcinoma in patients with hepatitis B virus-related liver cirrhosis[J].Journal of Tianjin Medical University,2017,23(01):214.
[2]周冷潇,韩 涛,刘 芳.无创肝纤维化指标结合甲胎蛋白对乙型肝炎相关肝细胞癌的评估[J].天津医科大学学报,2017,23(05):415.
　ZHOU Leng-xiao,HAN Tao,LIU Fang.Assessment of non-invasive fibrosis indexes with alpha-fetoprotein?for? hepatitis B virus-related hepatocellular carcinoma[J].Journal of Tianjin Medical University,2017,23(01):415.
[3]张自立,石文霞,李 霖,等.肝癌血清中miRNA-183的表达及临床意义[J].天津医科大学学报,2017,23(06):519.
　ZHANG Zi-li,?SHI Wen-xia,LI Lin,et al.Expression and significance of serum microRNA-183 in hepatocellular carcinoma[J].Journal of Tianjin Medical University,2017,23(01):519.
[4]侯振宇,孔银龙,孙 林,等.92例晚期肝细胞癌患者肝切除预后及危险因素分析[J].天津医科大学学报,2018,24(05):425.
　HOU Zhen-yu,KONG Yin-long,SUN Lin,et al.Prognosis and survival risk factors for 92 advanced hepatocellular carcinoma patients after hepatectomy[J].Journal of Tianjin Medical University,2018,24(01):425.
[5]胡 源,许戈良,荚卫东,等.C1QL1蛋白在原发性肝细胞癌中的表达及其临床意义[J].天津医科大学学报,2019,25(04):329.
　HU Yuan,XU Ge-liang,JIA Wei-dong,et al.Expressions of C1QL1 protein inprimaryhepatocellular carcinoma and its clinical significance[J].Journal of Tianjin Medical University,2019,25(01):329.
[6]张萃萃,邓为民.基因表达谱分析肝细胞癌的特征基因[J].天津医科大学学报,2020,26(06):514.
　ZHANG Cui-cui,DENG Wei-min.Gene expression profiling reveals important characteristic genes in hepatocellular carcinoma[J].Journal of Tianjin Medical University,2020,26(01):514.
[7]张杨,游阿彬,齐寒,等.负载化疗药物的外泌体对肝癌的靶向治疗研究[J].天津医科大学学报,2021,27(03):229.
　ZHANG Yang,YOU A-bin,QI Han,et al.Tumor-derived exosomes mediate targeted therapy in hepatocellular carcinoma mice[J].Journal of Tianjin Medical University,2021,27(01):229.
[8]王凤松,朱刘洋,白易,等.基于肿瘤突变负荷构建肝细胞癌风险评分预后模型[J].天津医科大学学报,2022,28(01):20.
　WANG Feng-song,ZHU Liu-yang,BAI Yi,et al.Identification of a risk score prognostic model of hepatocellular carcinoma based on tumor mutation burden[J].Journal of Tianjin Medical University,2022,28(01):20.
[9]赵耕,张盈莹,卓永,等.藏区慢性乙型肝炎患者使用PAGE-B模型对肝细胞癌的风险预测研究[J].天津医科大学学报,2022,28(06):654.
　ZHAO Geng,ZHANG Ying-ying,ZHUO Yong,et al.Risk prediction of hepatocellular carcinoma using the PAGE-B model in Tibetan patients with chronic hepatitis B[J].Journal of Tianjin Medical University,2022,28(01):654.
[10]元喆悦,白易,童文,等.基于COVID-19相关基因的肝细胞癌分子分型及预后模型构建与验证[J].天津医科大学学报,2024,30(01):15.[doi:10.20135/j.issn.1006-8147.2024.01.0015]
　YUAN Zheyue,BAI Yi,TONG Wen,et al.Molecular typing of hepatocellular carcinoma based on COVID-19 related genes and construction and validation of prognostic model[J].Journal of Tianjin Medical University,2024,30(01):15.[doi:10.20135/j.issn.1006-8147.2024.01.0015]

备注/Memo

备注/Memo:

作者简介 张华（1989-），男，主管技师，硕士在读，研究方向：免疫学；通信作者：张学军，E-mail：xjzh@tmu.edu.cn。

常用功能

更新日期/Last Update: 2024-01-01