[Effects of hepatocyte growth factor gene transfection on adriamycin-induced apoptosis of gastric cancer cells in vitro].
OBJECTIVE
To investigate the effects of hepatocyte growth factor (HGF) on adriamycin-induced apoptosis of gastric cancer cells in vitro.


METHODS
An 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.


RESULTS
The 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.


CONCLUSIONS
HGF gene transfection may suppress adriamycin-induced apoptosis of gastric cancer cells.
Shen
Jian-guo
JG
Department of Surgical Oncology, Sir Run Run Shaw Hospital, Zhejiang University College of Medicine, Hangzhou 310016, China.
Cheong
Jae-ho
JH
Noh
Sung-hoon
SH
Wang
Lin-bo
LB
D000903:Antibiotics, Antineoplastic / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects* / Q000235:genetics / Q000502:physiology; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D053938:DNA Fragmentation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004317:Doxorubicin / Q000494:pharmacology*; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D017228:Hepatocyte Growth Factor / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D010957:Plasmids / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013274:Stomach Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D014162:Transfection
D000903:Antibiotics, Antineoplastic; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D017228:Hepatocyte Growth Factor; D004317:Doxorubicin