Effect of heparanase on the adhesion and invasion ability of hepa-toma carcinoma cells
10.3969/j.issn.1000-8179.20141013
- VernacularTitle:肝素酶调节对肝癌细胞粘附侵袭能力的影响
- Author:
Xiaopeng CHEN
;
Junsheng LUO
;
Ye TIAN
;
Chenlin NIE
;
Wei CUI
;
Weidong ZHANG
- Publication Type:Journal Article
- Keywords:
hepatocellular carcinoma;
heparanase;
RNA interference;
vector;
cell adhesion;
invasion
- From:
Chinese Journal of Clinical Oncology
2014;(18):1180-1184
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the effect of heparanase (HPSE) on the cell adhesion and invasion ability of hepatoma carcino-ma (HC) cell. Methods:HPSE expressions in human HC cell lines (BEL-7402, HepG2, and HCCLM3) were measured by real-time re-verse transcriptase-polymerase chain reaction (RT-PCR) and Western blot analysis. Four recombinant miRNA vectors, pcD-NATM6.2-GW/EmGFP-miR-HPSE (pcDNA-miR-HPSE), were constructed and transfected into HCCLM3 cells. Full-length cDNA of HPSE gene was cloned into pIRES2-EGFP vector and transfected into HepG2 cells. Transfection efficiency was observed with fluores-cence microscope. HPSE expressions in transfected cells were measured by real-time RT-PCR and Western blot analysis. Adher-ence ability was determined with microplate reader, and invasion and migration abilities were detected with Transwell chambers. Re-sults:Both HPSE mRNA and protein relative expression levels were higher in the three types of HC cells than those in normal hepato-cyte (P<0.05). HPSE had the highest expression level in HCCLM3 cells and the lowest expression level in HepG2 cells (P<0.05). All five recombinant vectors met the experimental requirements. The transfection efficiencies were 75%-85%. The four miRNA vectors, pcDNA-miR-HPSE, significantly decreased HPSE expression in transfected HCCLM3 cells (P<0.05), and pcDNA-miR-HPSE-1 showed the best interference effect (P<0.05). Plasmid pIRES2-EGFP-HPSE increased HPSE expression in transfected HepG2 cells (P<0.05). After pcDNA-miR-HPSE-1 was transfected, the HCCLM3 cell adherence rate and the cell invasion and migration numbers dropped by almost 50%(P<0.01). After transfection of pIRES2-EGFP-HPSE, the HepG2 cell adherence rate and the cell invasion and migration numbers increased by nearly 40%(P<0.05). Conclusion:Different HPSE vectors could regulate bi-directionally the adher-ence, invasion, and migration abilities of transfected HC cells. HPSE may be related with adherence aside from invasion of HC cell.
