Inhibitory effects of c-erbB-2 antisense oligonucleotide transfection on uterine endometrial cancer Ishikawa cell lines.
BACKGROUND
C-erbB-2 is a chief proto-oncogene of endometrial cancer, which plays an important role in the biological behavior of endometrial cancer. Its overespression is an important factor of poor progrosis. The objective of this study was to investigate the treatment effects of transfecting c-erbB-2 antisense oligonucleotide (ASODN) on the uterine endometrial cancer Ishikawa cell line.


METHODS
The c-erbB-2 expression on Ishikawa cell membranes was determined by immunohistochemistry and then the aim was to transfect ASODN into Ishikawa cells and assay the cellular growth inhibition by MTT, to observe the cellular ultrastructure changes under transmission electron microscope (TEM), and to assay the cellular apoptotic rate, c-erbB-2 mRNA and protein expression by flow cytometry, RT-PCR and Western blot, respectively.


RESULTS
C-erbB-2 protein was positively expressed on Ishikawa cell membranes. MTT showed that when the concentration of transfecting ASODNs was 0.3 uM and 0.6 uM, cell growth inhibition rates were 55.43% and 76.12%, respectively. After transfecting 0.3 uM ASODN, the Ishikawa cell ultrastructure was obviously damaged, the cellular apoptotic rate was 72.21%, and the c-erbB-2 mRNA and protein expression were 45.71% and 34.52%, respectively, compared with those of the normal control cells.


CONCLUSIONS
Transfecting c-erbB-2 ASODN can obviously suppress its mRNA and protein expression in Ishikawa cells, cause cellular apoptosis and inhibit cell growth. It may have an important role in the gene therapy of endometrial cancer.
Zhao
F J
FJ
Department of Obstetrics and Gynecology, Shengjing Hospital, China Medical University, Shen Yang, PR China. zhaofujie63@yahoo.com.cn
Zhang
S L
SL
Ma
L
L
Gao
H
H
Zong
Z H
ZH