Screening and construction of hepatocellular carcinoma cell model regulated by GP73 expression

doi:10.3877/cma.j.issn.2095-5820.2021.01.002

Abstract

Abstract:

Objective

To find the appropriate cell lines to construct interference and overexpression of hepatoma cell model, so as to provide a suitable cell model for further study on the mechanism of GP73 regulation of hepatocellular carcinoma.

Methods

Eight hepatoma cell lines were selected including Huh-7、Hep3B、MHCC-LM3、MHCC97-L、MHCC97-H、SNU-449、SNU-398、SNU-182. The expression of GP73 was verified by q-PCR and Western Blot. According to the results, a hepatoma cell line was selected for interference and overexpression of GP73. The effects of overexpression of GP73 on the migration and invasion of selected cells were observed by Transwell experiments.

Results

Hep3B cell lines were selected for interference and overexpression of GP73. q-PCR and Western Blot results showed that the expression of GP73 in Hep3B cell line was significantly different from that in control group (P＜0.05).Transwell experiments indicated that overexpression of GP73 can significantly promote the migration and invasion of Hep3B cells.

Conclusion

The successful construction of Hep3B cell model provides a reliable research model for the mechanism of GP73 regulation of hepatocellular carcinoma, and provides a research basis for the following study.

Key words: Cell model, GP73, Primary liver cancer

Peiling Ye, Hongyun Jia, Liang Peng. Screening and construction of hepatocellular carcinoma cell model regulated by GP73 expression[J]. Chinese Journal of Clinical Laboratory Management(Electronic Edition), 2021, 09(01): 6-11.

/ / Recommend

Add to citation manager EndNote|Ris|BibTeX

URL: https://zhlcsysgldzzz.cma-cmc.com.cn/EN/10.3877/cma.j.issn.2095-5820.2021.01.002

https://zhlcsysgldzzz.cma-cmc.com.cn/EN/Y2021/V09/I01/6

Figures/Tables 9

References 13

1	Bary F, Ferlay J, Soerjomataram I, et al. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J]. ACJC, 2018, 68(6):394-424.
2	张宇龙. 基于RNA干扰技术沉默GP73基因对肝癌细胞增殖和凋亡影响的研究[D]. 兰州: 兰州大学, 2014.
3	叶佩灵, 何欣欣, 吴晓蔓. 原发性肝癌组织中GP73蛋白的表达[J]. 广东医学, 2014, 35(2):234-236.
4	叶佩灵, 吴晓蔓. 高尔基体糖蛋白73在原发性肝癌中的表达及意义[J]. 海南医学院学报, 2014, 20(1):83-86.
5	叶佩灵, 吴晓蔓. 血清GP73联合AFP、CEA、CA199、GGT检测在PHC早期筛查的应用价值[J]. 热带医学杂志, 2016, 16(4):467-470.
6	Ai N, Liu W, Li ZG, et al. High expression of GP73 in primary hepatocellular carcinoma and its function in the assessment of transcatheter arterial chemoembolization[J]. Oncol Lett, 2017, 14(4):3953-3958.
7	Kladney RD, Bulla GA, Guo Linsheng, et al. GP73, a novel golgi-localized protein upregulated by viral infection [J]. Gene, 2000, 249(1-2):53-65.
8	Wright LM, Huster D, Lutsenko S, et al. Hepatocyte GP73 expression in Wilson disease[J]. J Hepatol, 2009, 51(3):557-564.
9	王小华, 相祎, 王伟. 肝细胞肝癌血清GP73和HMGB1的表达水平及其联合检测意义[J]. 肝脏, 2018, (10):914-917.
10	Glenn S, Peter B, Craig WG, et al. Identification of novel clinical factors associated with hepatic fat accumulation in extreme obesity[J]. J Obes, 2014(2014):368210.
11	Yang Y, Liu Q, Li ZP, et al. GP73 promotes epithelial-mesenchymal transition and invasion partly by activating TG-β1/Smad2 signaling in hepatocellular carcinoma[J]. Carcinogenesis, 2018, 39(7):900-910.
12	陈默, 赵坤, 柳鹏程. RNA干扰技术沉默GP73对人肝癌HepG2细胞迁移和侵袭的影响[J]. 上海交通大学学报, 2016(36), 11:1588-1593.
13	刘丽萍, 陈健康, 刘元明.高尔基体蛋白73在肝细胞癌转移及侵袭中的作用[J]. 中华肿瘤杂志, 2017, 39(7):497-501.

基因名称	序列
基因名称	sense(5'-3')	antisense(5'-3')
si-GP73	CAAGCUGUACCAGGACGAATT	UUCGUCCUGGUACAGCUUGTT
siRNA NC	CCAUAAGGACGUCAGACGATT	UCGUCUGACGUCCUUATGGTT

基因名称	序列
基因名称	sense(5'-3')	antisense(5'-3')
si-GP73	CAAGCUGUACCAGGACGAATT	UUCGUCCUGGUACAGCUUGTT
siRNA NC	CCAUAAGGACGUCAGACGATT	UCGUCUGACGUCCUUATGGTT

基因名称	Primer名称	序列(5'-3')	产物长度(bp)
GP73	GP73-F	GCCGGAGCCTCGAAAAGAG	190
GP73	GP73-R	TTCCAGCTCCATGATCCGTG	190
GAPDH	GAPDH-F	GAGTCAACGGATTTGGTCGT	185
GAPDH	GAPDH-R	GACAAGCTTCCCGTTCTCAG	185

基因名称	Primer名称	序列(5'-3')	产物长度(bp)
GP73	GP73-F	GCCGGAGCCTCGAAAAGAG	190
GP73	GP73-R	TTCCAGCTCCATGATCCGTG	190
GAPDH	GAPDH-F	GAGTCAACGGATTTGGTCGT	185
GAPDH	GAPDH-R	GACAAGCTTCCCGTTCTCAG	185

[1]	Chuanju Li, Linyue Liu, Mei Wang, Xin Li, Xianghui Han, Haiyong Jia. Progress on models of hepatitis B virus infection [J]. Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition), 2022, 16(06): 361-365.
[2]	Zhengju Xu, Houquan Hu, Dandan Xie, Xiaoman Zhang, Liguan Liu, Lianqiu Zheng, Yanyan Yan, Zhijie Huang. Changes of serum Golgi protein 73 of patients with influenza [J]. Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition), 2019, 13(03): 190-195.
[3]	Yaobo Xu, Binquan Wu. Application of three-dimensional visualization combined with intraoperative ultrasound in laparoscopic surgery for resectable primary liver cancer [J]. Chinese Archives of General Surgery(Electronic Edition), 2022, 16(04): 273-277.
[4]	Baofu Zhou, Lele Wu, Yonghong Li, Shichao Hu. Effect of different liver cutting methods in hepatectomy for multifocal primary liver cancer [J]. Chinese Archives of General Surgery(Electronic Edition), 2022, 16(03): 205-209.
[5]	Qian Yang, Cuifang Li, Wanqiu Zhang. Analysis of short-term and long-term prognosis and influencing factors of primary liver cancer with spontaneous ruptured hemorrhage after emergency TACE surgery [J]. Chinese Journal of Operative Procedures of General Surgery(Electronic Edition), 2024, 18(01): 33-36.
[6]	Yiqi Dong, Yaobing Luo, Tao Liu, Lan Zhang, Ming Yang. Application value of blood flow topology liver segmentation and Couinaud liver segmentation in preoperative evaluation and surgical planning of primary liver cancer [J]. Chinese Journal of Operative Procedures of General Surgery(Electronic Edition), 2022, 16(04): 423-426.
[7]	Yafeng Chen, Jiangbin Li, Wenjun Zhong, Rui Dong, Li Zang, Jikai Yin, Xilin Du, Jianguo Lu. Clinical effect analysis of laparoscopic microwave ablation for HCC ≤ 3 cm [J]. Chinese Journal of Laparoscopic Surgery(Electronic Edition), 2021, 14(05): 271-276.
[8]	Bing Han, Jinyang Gu. Research and application prospect of deep learning neural network in diagnosis and treatments for liver cancer [J]. Chinese Journal of Hepatic Surgery(Electronic Edition), 2023, 12(05): 480-485.
[9]	Chufeng Wang, An Jiang. Molecular diagnosis of primary liver cancer [J]. Chinese Journal of Hepatic Surgery(Electronic Edition), 2023, 12(05): 499-503.
[10]	Rong He, Zhe Jia, Ke Zhang, Daijing Li, Meng Zhang, Li Jiang. Analysis of clinical efficacy of laparoscopic hepatectomy combined with Hassab's procedure for primary liver cancer complicated with portal hypertension based on propensity score matching [J]. Chinese Journal of Hepatic Surgery(Electronic Edition), 2023, 12(04): 376-383.
[11]	Yunpeng Ge, Hongyuan Cui, Jinghai Song. Application of artificial intelligence in diagnosis, treatment and prognosis of primary liver cancer [J]. Chinese Journal of Hepatic Surgery(Electronic Edition), 2023, 12(04): 367-371.
[12]	Jinyu Jiang, Donglei Zhang, Wei He. Advances in the application of blue light induced-oxidative damage models in the study of ophthalmic diseases [J]. Chinese Journal of Ophthalmologic Medicine(Electronic Edition), 2022, 12(03): 168-172.
[13]	Lan Wu, Jianyu Feng. Effect of lidocaine intransarterial chemoembolization for liver cancer [J]. Chinese Journal of Interventional Radiology(Electronic Edition), 2022, 10(03): 333-338.
[14]	Hui Li, Haixia Li, Xiuxiang Wang. Efficacy of Xuebijing injection in the treatment of post-interventional Syndrome of primary liver cancer and its effects on liver function and serum CRP, TNF-AND VEGF levels [J]. Chinese Journal of Interventional Radiology(Electronic Edition), 2022, 10(01): 27-31.
[15]	Qin Shi, Chen Zhou, Jiacheng Liu, Songjiang Huang, Chongtu Yang, Bin Xiong. Application of drug-loaded embolic agents in the treatment of hepatocellular carcinoma [J]. Chinese Journal of Interventional Radiology(Electronic Edition), 2020, 08(04): 364-369.

Please choose a citation manager

Content to export