Antisense oligonucleotides targeting midkine induced apoptosis and increased chemosensitivity in hepatocellular carcinoma cells

Abstract

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 antitumor activity. 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 as apoptosis and caspase‐3 activity were also examined in HepG2 cells. Cell proliferation was then analyzed after treatment with both ASODN and chemotherapeu‐tic drugs. Results: In this experiment, the ASODN5 among the 10 ASODN showed higher inhibitory activity against proliferation of hepatocellular carcinoma cells in a dose‐dependent manner. In HepG2 cells, ASODN5 could significantly reduce the MK mRNA level and protein content. After transfection with ASODN5 for 48 h, accompanied with a decline of survivin and Bcl‐2 protein content, a remarkable increase of apoptosis and caspase‐3 activity was observed in HepG2 cells. Furthermore, ASODN5 transfer can significantly increase chemosensitivity in HepG2 cells. Conclusion: Antisense oligonucleotides targeting MK shows therapeutic effects on HCC; ASODN5 has the possibility to be developed as an effective antitumor agent.

Antisense oligonucleotide targeting midkine suppressesin vivoangiogenesis

“…Additionally, MK, an angiogenic factor, is expressed in bladder cancer, www.wjgnet.com and its over-expression correlates with a poor outcome in patients with invasive cancers [20] . In fact, we have confirmed that MK-AS transfer can significantly inhibit the growth of hepatocellular carcinoma cells, which is associated with increased Caspase-3 activity [34] . The present study was to determine whether MK-AS can suppress angiogenesis, which is an important mechanism underlying inhibition of tumor cell proliferation.…”

Antisense oligonucleotides targeting midkine inhibit tumor growth in an in situ human hepatocellular carcinoma model

“…In fact, several studies have been reported that antisense targeting MK inhibits tumor growth, such as human prostate cells, colon carcinoma cells [26] , and mouse rectal carcinoma cells [27] . We previously confirmed that MK-AS could significantly inhibit growth of hepatocellular cells including HepG2, SMMC-7721, and BEL-7402 [28] . The aim of this study was to evaluate the in vivo antitumor effects of MK-AS in an in situ human hepatocellular carcinoma model in mice.…”

In vitro and in vivo suppression of hepatocellular carcinoma growth by midkine-antisense oligonucleotide-loaded nanoparticles

“…Recently, HCC tumor cells were found to overexpress MK. In our previous studies, ASODNs that target MK were demonstrated to play an important role in anti-tumor functions [15,16] . However, the anti-tumor effect was not satisfactory and found to be toxic because of the absence of smart and safer delivery tools.…”

“…In our previous study [15] , this antisense sequence has been identified to be the most effective sequence for the down-regulation of MK. Consequently, this sequence was synthesized and applied in this study.…”