The synergism and mechanism of action of rClone30-hDR5 in combination with TRAIL on HCC.
- Author:
Tian SUN
;
Ze-Shan NIU
;
Xue-Ying LIU
;
Gui-You TIAN
;
Yin BAI
;
Fu-Liang BAI
;
Jie-Chao YIN
;
Dan YU
;
Yun-Zhou WU
;
De-Shan LI
;
Qing-Zhong YU
;
Si-Ming LI
;
Gui-Ping REN
- Publication Type:Journal Article
- MeSH:
Apoptosis;
Carcinoma, Hepatocellular;
pathology;
Caspase 3;
metabolism;
Caspase 8;
metabolism;
Drug Synergism;
Hep G2 Cells;
Humans;
Liver Neoplasms;
pathology;
Real-Time Polymerase Chain Reaction;
Receptors, TNF-Related Apoptosis-Inducing Ligand;
pharmacology;
TNF-Related Apoptosis-Inducing Ligand;
pharmacology;
Transfection
- From:
Acta Pharmaceutica Sinica
2014;49(7):985-992
- CountryChina
- Language:Chinese
-
Abstract:
To investigate the cell-killing effect and its possible mechanism of rClone30-hDR5 in combination with TRAIL on human hepatic carcinoma (HCC) cell line, first of all, recombinant plasmid pee12.4-hDR5 was introduced into HepG2 cells by liposome transfection. After five rounds of screening by flow cytometry, HepG2 cells expressing high levels of DR5 on cell surface were isolated. The cytotoxicity of TRAIL to selected cells was higher than that of TRAIL to HepG2 cells by MTT method (P < 0.01). The result suggested that the cloned hDR5 gene had biological activity. MTT assay showed that, rClone30- hDR5 in combination with TRAIL more efficiently inhibited the tumor growth of HepG2 cells compared to rClone30-hDR5 or TRAIL in vitro. The results of Annexin V-FITC/PI staining and Quantitative Real-time PCR indicated that rClone30-hDR5 in combination with TRAIL significantly increased the mRNA levels of caspase 3 and caspase 8, and induced the apoptosis of tumor cells. HepG2 cells were infected with rClone30-hDR5 or rClone30 at MOI of 1. The expression of hDR5 on tumor surface increased significantly by rClone30-hDR5 compared to that by rClone30, which contributed to the sensitivity to TRAIL. In conclusion, rClone30-hDR5 in combination with TRAIL has potential application value in cancer treatment.
