ITIH4基因沉默对人肝癌细胞HepG2增殖、迁移及侵袭的影响
| 作者: |
1段煜,
1姜文文,
2高鹏,
3唐泽耀
1 大连医科大学附属第六人民医院肝病科，辽宁 大连 116044 2 大连医科大学附属第六人民医院检验科，辽宁 大连 116044 3 大连医科大学药理学教研室，辽宁 大连 116044 |
| 通讯: |
段煜
Email: duanyu_6hospital@163.com |
| DOI: | 10.3978/j.issn.2095-6959.2017.07.001 |
| 基金: | 辽宁省自然科学基金, 201602206 |
摘要
目的：探讨沉默间-α-胰蛋白酶抑制剂重链H4(inter-alpha-trypsin inhibitor heavy chain H4，ITIH4)基因对人肝癌细胞系HepG2增殖、迁移及侵袭的影响。方法：将4个ITIH4基因沉默载体分别转染HepG2细胞，用2 mg/L嘌呤霉素筛选获得稳定表达细胞株。Western印迹评价沉默效率，并选择效率最高的细胞株(si-ITIH4)用于后续实验。转染无意义序列的细胞株(si-Control)作为对照组。分别用细胞计数及CCK-8细胞计数试剂盒评价两组细胞增殖。用Transwell小室评价两组细胞的迁移及侵袭。结果：成功获得ITIH4基因沉默的HepG2细胞株si-ITIH4，与si-Control组相比，si-ITIH4组沉默效率达到86%。与si-Control组相比，si-ITIH4组细胞增殖显著降低、细胞迁移及侵袭均显著减少，差异有统计学意义(P<0.05)。结论：ITIH4基因沉默可显著抑制HepG2的增殖、迁移及侵袭，提示其可能是参与肝细胞癌发生发展的重要基因之一。
关键词：
间-α-胰蛋白酶抑制剂重链H4
增殖
迁移
侵袭
肝细胞癌
Effect of ITIH4 gene silence on proliferation, migration and invasion of human hepatocellular carcinoma cell HepG2
CorrespondingAuthor: DUAN Yu Email: duanyu_6hospital@163.com
DOI: 10.3978/j.issn.2095-6959.2017.07.001
Abstract
Objective: To investigate the effect of inter-alpha-trypsin inhibitor heavy chain H4 gene silence on proliferation, migration and invasion of human hepatocellular carcinoma cell HepG2. Methods: Four ITIH4 silencing vectors were delivered in to HepG2 cells by transfection. Stable cell clones were selected by 2 μg/mL puromycin. Silencing efficiency was evaluated by Western Blot and cell clone with the highest silencing efficacy (si-ITIH4) was used in the following study. Cell clone transfected with scramble sequence was used as normal control (si-Control). Cell counting and CCK-8 assay were applied to assess cell proliferation. Transwell assay was performed to evaluate cell migration and invasion. Results: We successfully obtained a HepG2 cell line with ITIH4 gene silencing named si-ITIH4. The silencing efficiency was 86% in compassion with si-Control. Compared to the si-Control group, si-ITIH4 cells showed significantly slower proliferation, decreased migration and mitigated invasion. Conclusion: Silence of ITIH4 significantly inhibited proliferation, migration and invasion of HepG2 cells. This study suggests that ITIH4 might be essentially involved in onset and progression of hepatocellular carcinoma.