ZNF384调控GCLM对肝细胞癌转移的机制研究

张明; 于成龙; 曹锦鹏

doi:10.12259/j.issn.2095-610X.S20231108

ZNF384调控GCLM对肝细胞癌转移的机制研究

doi: 10.12259/j.issn.2095-610X.S20231108

1.
青岛市市南区人民医院检验科
2.
青岛市第三人民医院健康管理中心，山东青岛　266000

基金项目: 青岛市输血协会科技计划项目（2020-qdsx11）

详细信息

作者简介:
张明（1971～），男，山东青岛人，医学学士，副主任技师，主要从事临床基础检验工作

通讯作者:
曹锦鹏，E-mail：epeng12345@sina.com

中图分类号: R735.7
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- 被引次数: 0
出版历程
- 收稿日期: 2023-09-05
- 网络出版日期: 2023-11-04
- 刊出日期: 2023-11-30

ZNF384 Regulates Metastasis of Hepatocellular Carcinoma via GCLM

1.
Dept. of Clinical Laboratory，Shinan District People’s Hospital of Qingdao
2.
Health Management Center Affiliated Qingdao Third People’s Hospital，Qingdao University，Shandong Qingdao 266000，China

摘要

摘要: 目的探讨ZNF384通过调控GCLM对肝细胞癌（hepatocellular carcinoma，HCC）增殖和转移的影响及具体调控机制。方法分别在HCC细胞系HepG2和HuH7中转染si-ZNF384和pcDNA-GCLM。Western blot检测ZNF384和GCLM蛋白表达以及上皮间质转化（epithelial-mesenchymal transition，EMT）标志物N-cadherin、Vimentin、E-cadherin的表达，CCK-8检测细胞增殖活力，Transwell检测细胞迁移。双荧光素酶报告基因实验验证ZNF384和GCLM结合关系。荧光原位杂交（fluorescence in situ hybridization，FISH）技术检测ZNF384和GCLM在HepG2细胞中的定位。结果 ZNF384和GCLM在HepG2和HuH7细胞中表达升高（P < 0.05），敲降ZNF384抑制HepG2和HuH7细胞增殖（P < 0.001）、迁移（P < 0.001）以及间质标志物N-cadherin（P < 0.01）和Vimentin（P < 0.01）的表达，促进上皮标志物E-cadherin的表达（P < 0.01）。数据库以及双荧光素酶实验证实ZNF384与GCLM启动子结合。过表达GCLM逆转敲降ZNF384对HepG2和HuH7细胞增殖、迁移以及EMT的抑制作用（P < 0.05）。结论敲降ZNF384通过抑制GCLM，抑制HCC细胞增殖和转移。
- 肝细胞癌 /
- ZNF384 /
- GCLM /
- 增殖 /
- 转移
Abstract: Objective To investigate the effect of ZNF384 on the proliferation and metastasis of HCC by regulating GCLM and demonstrate the specific mechanism. Methods si-ZNF384 and pcDNA-GCLM were transfected into HepG2 and HuH7 cell lines of HCC, respectively. Western blot was carried out to detect the expressions of ZNF384 and GCLM proteins as well as the expressions of EMT-markers N-cadherin, Vimentin and E-cadherin. CCK-8 assessed cell proliferative activity. Transwell assay was conducted to measure cell migration. Dual-luciferase experiment verified the binding relationship between ZNF384 and GCLM. The localization of ZNF384 and GCLM in HepG2 cells was detected by FISH assay. Results The expressions of ZNF384 and GCLM in HepG2 and HuH7 cells were increased（P < 0.05）. Knockdown of ZNF384 inhibited the proliferation（P < 0.001） and migration（P < 0.001） of HepG2 and HuH7 cells, suppressed the expression of interstitial markers N-cadherin（P < 0.01） and Vimentin（P < 0.01）, and promoted the expression of epithelial marker E-cadherin（P < 0.01）. Database and dual-luciferase experiments confirmed that ZNF384 is bound to the promoter of GCLM. Overexpression of GCLM reversed the inhibitory effect of knocking down ZNF384 on the proliferation, migration and EMT of HepG2 and HuH7 cells（P < 0.05）. Conclusion Knocking down ZNF384 can repress the proliferation and metastasis of HCC cells by inhibiting GCLM.
- Hepatocellular carcinoma /
- ZNF384 /
- GCLM /
- Proliferation /
- Metastasis

HTML全文

图 1 敲降ZNF384抑制HCC细胞增殖和转移

A～B：Western blot检测HCC细胞系中ZNF384蛋白表达，与LO2组相比，^*P < 0.05，^***P < 0.001。C～E：Western blot检测敲降ZNF384对ZNF384蛋白表达的作用；F：CCK-8检测细胞增殖活力；G～H：Transwell检测细胞迁移能力；I-K：Western blot检测EMT相关标志物的表达。与si-NC组相比，^*P < 0.05，^**P < 0.01，^***P < 0.001。

Figure 1. Knockdown of ZNF384 inhibited the proliferation and metastasis of HCC cells

下载: 全尺寸图片幻灯片

图 2 ZNF384作为转录因子调控GCLM的表达

A：AnimalTFD3.0数据库预测ZNF384与GCLM的启动子结合位点;B：双荧光素酶报告基因实验验证ZNF384与GCLM的结合关系，与pcDNA-NC组相比，^***P < 0.001；C：FISH检测ZNF384和GCLM在HepG2细胞中的定位；D：Western blot检测HCC细胞中GCLM的蛋白表达，与LO2组相比，^**P < 0.01，^***P < 0.001；E～F：Western blot检测敲降ZNF384对GCLM蛋白表达的作用。与si-NC组相比，^**P < 0.01，^***P < 0.001。

Figure 2. ZNF384 as a transcription factor regulated the expression of GCLM

下载: 全尺寸图片幻灯片

图 3 ZNF384调控GCLM抑制肝细胞癌细胞的增殖和转移

A～C：Western blot检测ZNF384和GCLM在HepG2和HuH7细胞中的表达；D：CCK-8检测细胞增殖活力；E～G：Transwell检测细胞迁移能力；H～J：Western blot检测EMT相关标志物N-cadherin、Vimentin和E-cadherin的蛋白表达。^*P < 0.05，^**P < 0.01，^***P < 0.001。

Figure 3. ZNF384 suppressed the proliferation and metastasis of HCC cells by regulating GCLM

下载: 全尺寸图片幻灯片

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