Effects of hepatocyte growth factor gene transfection on adriamycin-induced apoptosis of gastric cancer cells in vitro.
- Author:
Jian-guo SHEN
1
;
Jae-ho CHEONG
;
Sung-hoon NOH
;
Lin-bo WANG
Author Information
- Publication Type:Journal Article
- MeSH: Antibiotics, Antineoplastic; pharmacology; Apoptosis; drug effects; genetics; physiology; Cell Line, Tumor; Cell Proliferation; drug effects; Cell Survival; drug effects; genetics; physiology; DNA Fragmentation; drug effects; Dose-Response Relationship, Drug; Doxorubicin; pharmacology; Flow Cytometry; Green Fluorescent Proteins; genetics; metabolism; Hepatocyte Growth Factor; genetics; metabolism; physiology; Humans; Plasmids; genetics; RNA, Messenger; genetics; metabolism; Recombinant Fusion Proteins; genetics; metabolism; Stomach Neoplasms; genetics; metabolism; pathology; Transfection
- From: Chinese Journal of Oncology 2007;29(5):338-341
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the effects of hepatocyte growth factor (HGF) on adriamycin-induced apoptosis of gastric cancer cells in vitro.
METHODSAn eukaryotic expression plasmid (pIRES2-EGFP) containing HGF (pIRES2-EGFP-HGF) was constructed. Human gastric cancer cell line MKN-45 cells were transfected in vitro with pIRES2-EGFP containing HGF or not. RT-PCR and Western blot were used to determine the target gene expression. Function of HGF was determined by MDCK cell scattering assay. Cell viability was tested by MTT assay. Apoptosis was evaluated by DNA fragmentation assay as well as flow cytometry using PI staining.
RESULTSThe HGF transfected MKN45 cells could stably express HGF mRNA, and secrete HGF protein to the cell culture median which was detected to exhibit normal function. The cell inhibition rate induced by adriamycin in HGF-transfected cells was decreased as compared to that of parental and mock transfected cells. When treated with adriamycin at 0.1 microg/ml, the parental and mock transfected cells present typical apoptotic ladder on DNA electrophoresis while HGF transfected cell did not. The apoptotic rate was decreased in HGF transfected cells as compared with that of parental and mock transfected cells.
CONCLUSIONHGF gene transfection may suppress adriamycin-induced apoptosis of gastric cancer cells.
