| BackgroundMost tumors accompany with gene mutation and/or overexpression,which result in the activation of oncogene,this is a very important process during the pathogenesis of the tumors.Thus the intent effectively silence the gene of interest by preventing synthesis of the protein that it encodes is'nonetheless attractive because mRNA is much more accessible and is efficient to be manipulated than DNA.At present,a large body of studies have focused on destabilizing mRNA directing at various targets that play a role in cancer,cardiovascular disease,viral disease,and inflammatory,accompanying with gene mutation and/or overexpression. Antisense technique,which have the characteristics of high selectivity and affinity to its targets,quickly taking effects,relatively definite mechanism,to be easy to evaluate the destination and less side effects,etc,is a popular method to effectively silence the gene now.Many satisfactory results have been gained on breast cancer,ovarian cancer,etc.While this kind of study on uterine endometrial cancer has not been reported up to now,and we have learned that c-erbB-2 is a chief proto-oncogene of endometrial cancer,which plays an important role in the biological behavior of endometrial cancer,and its overespression is an important factor of bad progrosis.Therefor the objective of this study is to investigate the treating effects of transfecting c-erbB-2 antisense oligonucleotides (ASODNs) on high differentiated and mid differentiated uterine endometrial cancer ISHIKAWA and HEC-1A cell line. Methods(1).To assay the c-erbB-2 protein expression on endometrial cander cells,and determine the relationship between its expression and clinical-pathological factors of endometrial cancer.(2).to determine that the c-erbB-2 is positively expressed on ISHIKAWA and HEC-1A cells membrane by S-P immunohistoche -mestry,which is a prerequisite of this study.(3).to transfect 0.1uM～0.6uM ASODNs into ISHIKAWA and HEC-1A cells,then to observe the cellular morphologic changes and to assay the cellular growth inhibition by MTT.(4).to transfect 0.3uM ASODNs,and to observe the cellular ultrastructure changes,to assay the cellular apoptotic rate,c-erbB-2 mRNA and protein expression by flow cytometry,TR-PCR and western blot respectively.Results(1).There is positive correlation between the c-erbB-2 protein expression and the histological grade of endometrial cancer.(2).c-erbB-2 protein was positively expressed on ISHIKAWA cells'membrane.(3).After transfecting 0.1uM～0.6uM ASODNs,the ISHIKAWA and HEC-1A cells had more and more obvious morphologic changes,such as shrinkage,damage and disintegration. MTT showed that when the concentration of transfecfing ASODNs was 0. 3uM and 0.6uM,the cells growth inhibition rate was 55.43%and 75.12%respectively. (4).After transfecting 0.3uM ASODNs,The ISHIKAWA cells had obvious vacuolar degeneration in their plasmas,there were disappearances of organelles and nuclear structure,and there was also obvious shrinkage of nucleus under transmission electron microscope(TEM).The cellular apoptotic rate was 72.21%.The c-erbB-2 mRNA and protein expression is 45.71%and 34. 52%respectively compared with the normal control cells. Conclusions(1).There is positive correlation between the c-erbB-2 protein expression and the histological grade of endometrial cancer.(2).With the increasing of the concentration of transfecting c-erbB-2 ASODNs,the ISHIKAWA and HEC-1A cells had more and more obvious morphologic changes and ultrastructure damages.(3).Transfecting c-erbB-2 ASODNs can obviously suppress its mRNA and protein expression in ISHIKAWA and HEC-1A cells.(4).Transfecting c-erbB-2 ASODNs can cause ISHIKAWA and HEC-1A cellular apoptosis. (5).With the increasing of the concentration of transfecting c-erbB -2 ASODNs,the inhibition of ISHIKAWA and HEC-1A cells growth were also strengthened.Thus it may suggested that the transfecting c-erbB-2 ASODNs may have an important role in the gene therapy of endometrial cancer.
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