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Aim:Overexpression of midkine(MK)has been observed in many malignancies.This aim of this study is to screen for suitable antisense oligonucleotides(ASODN)targeting MK in hepatocellular carcinoma(HCC)cells and evaluate its antitumoractivity.Methods:Ten ASODN targeting MK were designed and synthesized.After transfection with ASODN,cell proliferation was analyzed with MTS[3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium,inner salt]assay.In addition,MK mRNA,protein levels,as well asapoptosis and caspase-3 activity were also examined in HepG2 cells.Cell prolif-eration was then analyzed after treatment with both ASODN and chemotherapeu-tic drugs.Results:In this experiment,the ASODN5 among the 10 ASODN showedhigher inhibitory activity against proliferation of hepatocellular carcinoma cells ina dose-dependent manner.In HepG2 cells,ASODN5 could significantly reducethe MK mRNA level and protein content.After transfection with ASODN5 for48h,accompanied with a decline of survivin and Bcl-2 protein content,a remark-able increase of apoptosis and caspase-3 activity was observed in HepG2 cells.Furthermore,ASODN5 transfer can significantly increase chemosensitivity inHepG2 cells.Conclusion:Antisense oligonucleotides targeting MK shows thera-peutic effects on HCC;ASODN5 has the possibility to be developed as an effec-tive antitumor agent.
Aim: Overexpression of midkine (MK) has been observed in many malignancies. This aim of this study is to screen for suitable antisense oligonucleotides (ASODN) targeting MK in hepatocellular carcinoma (HCC) cells and evaluate its antitumoractivity. Methods: Ten ASODN targeting MK were designed and synthesized. After transfection with ASODN, cell proliferation was analyzed with MTS [3- (4,5-dimethylthiazol-2-yl) -5- (3-carboxymethoxyphenyl) -2- (4-sulfophenyl) , inner salt] assay.In addition, MK mRNA, protein levels, as well as caspase-3 activity were also examined in HepG2 cells. Cell prolif-eration was then analyzed with treatment with both ASODN and chemotherapeu- tic drugs. Results: In this experiment, the ASODN5 among the 10 ASODN showed hgher inhibitory activity against proliferation of hepatocellular carcinoma cells ina dose-dependent manner.In HepG2 cells, ASODN5 could significantly reduce the MK mRNA level and protein content. After transfection with ASODN5 for 48h, accompanied with a decline of survivin and Bcl-2 protein content, a remark-able increase of apoptosis and caspase-3 activity was observed in HepG2 cells. Frther and ASODN5 transfer can promote increase chemosensitivity inHepG2 cells. Confluence: Antisense oligonucleotides targeting MK shows thera-peutic effects on HCC; ASODN5 has the possibility to be developed as an effec-tive antitumor agent.