Construction of recombinant plasmids Egr1-XPO4 and its synergic inhibition with 5-FU against hepatocarcinoma SK-Hep1 cells
10.3872/j.issn.1007-385x.2018.02.004
- VernacularTitle:重组诱导性质粒Egr1-XPO4的构建及其与5-FU联合对肝癌细胞SKHep1的协同抑制作用
- Author:
WU Nan
1
,
2
;
HUANG Jing
3
,
4
;
CHEN Yang
5
;
LIU Kan
3
,
4
;
LIU Hao
3
,
4
;
WANG Xiaomei
3
,
4
Author Information
1. Department of Medicine, Shenzhen University, Shenzhen 518000，Guangdong,China;
2. Disease Control and Prevention Center of Nanshan District, Shenzhen 518000,Guangdong, China
3. (
4. Department of Medicine, Shenzhen University, Shenzhen 518000，Guangdong,China
5. China Life Health Industry Group, Shenzhen 518000, Guangdong, China
- Publication Type:Journal Article
- Keywords:
hepatocarcinoma;
SK-Hep1cell;
XPO4 gene;
Egr1 promoter;
5-FU;
synergic effect
- From:
Chinese Journal of Cancer Biotherapy
2018;25(2):132-136
- CountryChina
- Language:Chinese
-
Abstract:
[Abstract] Objective: To construct recombinant plasmid Egr1-XPO4 and evaluate its synergic inhibition with 5-FU against hepatocarcinoma SK-Hep1 cells. Methods: The XPO4 gene was inserted into vector carrying promoter Egr1 to construct a new recombinant vector, Egr1-XPO4, which was then transfected into human hepatocarcinoma cell line SK-Hep1 and sensitized with chemotherapeutic drug 5-FU. Western blotting was adopted to examine the protein expression of XPO4; CCK assay was used to detect SK-Hep1 cell proliferation after transfection, and Flow Cytometry with Annexin V-FITC/PI staining was used to detect the apoptosis of SK-Hep1 cells. SKHep1 cell xenograft model was constructed on nude mice, and the effect of Egr1-XPO4 in combination with 5-FU on the growth of xenograft was observed. Results: The recombinant plasmid Egr1-XPO4 was successfully constructed.With the sensitization of 5-FU, the expression of XPO4 protein in SK-Hep1 cells was significantly elevated after Egr1-XPO4 transfection, and the evlevation was in a 5FU dose-depend manner.The combined treatment of Egr1-XPO4 and 5-FU produced a significantly stronger inhibition against SKHep1 cell proliferation and greatly promoted apoptosis of SK-Hep1cells compared with 5-FU or pEgr-XPO4 mono-treatment group (all P<0.05). And in vivo antitumor experiment showed that the tumor volume in Egr1-XPO4+5-FU treatment group was significantly smaller than that of Egr1-XPO4 or 5-FU mono-treatment group (P<0.05). Conclusion: The recombinant plasmid Egr1-XPO4 in combination with 5-FU could exertsynergic inhibitionagainst hepatocarcinomaSK-Hep1 cells.
- Full text:20180204.pdf
