Antisense oligonucleotides targeting midkine inhibit tumor growth in an in situ human hepatocellular carcinoma model.
OBJECTIVE
To evaluate the in vivo antitumor effects of antisense oligonucleotides targeting midkine (MK-AS).


METHODS
An in situ human hepatocellular carcinoma (HCC) model was established in mice livers orthotopically. The MK-AS and 5- fluorouracil (5-Fu) were administered intravenously. The tumor sizes and plasma alpha-fetoprotein (AFP) were measured by calipers and radiation immunoassay respectively. The morphology of tumors was evaluated by hematoxylin-eosin staining of histological sections. Human MK, p53, Bax, Bcl-2, and caspase-3 protein content were detected by Western blotting.


RESULTS
MK-AS significantly inhibited in situ human HCC growth in mice compared with the saline group in a dose-dependent manner. After the treatment with MK-AS or with 5-Fu, the plasma AFP concentration decreased in a dose-dependent manner. Interestingly, MK-AS also clearly downregulated the protein level of Bcl-2, and upregulated p53, Bax, and caspase-3 in the hepatocellular carcinoma tissue.


CONCLUSIONS
These results demonstrated that MK-AS was an effective antitumor antisense oligonucleotide in vivo in mice; its antitumor effect is associated with the increase of pro-apoptotic proteins, such as p53, Bax, and caspase-3, and the decrease of the anti-apoptotic protein, Bcl-2.
Dai
Li-Cheng
LC
Huzhou Key Laboratory of Molecular Medicine, Huzhou Central Hospital, Huzhou 313000, China. dlc@hzhospital.com
Wang
Xiang
X
Yao
Xing
X
Min
Li-Shan
LS
Ping
Jin-Liang
JL
He
Jian-Fang
JF
D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D051017:Apoptosis Regulatory Proteins / Q000096:biosynthesis / Q000235:genetics; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy* / Q000473:pathology; D016207:Cytokines / Q000187:drug effects*; D006801:Humans; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D000078792:Midkine; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D023041:Xenograft Model Antitumor Assays; D000509:alpha-Fetoproteins / Q000096:biosynthesis / Q000235:genetics
D000970:Antineoplastic Agents; D051017:Apoptosis Regulatory Proteins; D016207:Cytokines; D016376:Oligonucleotides, Antisense; D000509:alpha-Fetoproteins; D000078792:Midkine