[Inhibition of invasiveness of human mammary carcinoma cell line MDA435 by heparanase antisense oligodeoxynucleotide].
OBJECTIVE
To evaluate the inhibitory effect of heparanase antisense oligodeoxynucleotide (AS-ODN) on the invasiveness of human mammary carcinoma cell line MDA435.


METHODS
The AS-ODN complementary to the start codon region of heparanase mRNA and its control, scrambled nonsense oligodeoxynucleotide (NS-ODN) were designed and synthesized and phosphorothioated. The ODNs were embedded in cationic liposome Lipofectin and transfected into MDA435 cells. The total RNAs and proteins were extracted from the cells 48 hours after transfection and then semi-quantitative RT-PCR and Western blotting were performed to evaluate the heparanase gene and protein expression levels respectively. The invasiveness of transfected MDA435 cells were measured quantitatively by Matrigel invasion assays.


RESULTS
The heparanase gene and protein expression and invasiveness of MDA435 cells treated with AS-ODN of different final concentrations were significantly decreased compared with that of the controls (P < 0.01). Besides, the inhibitory effects were significantly different between the cells treated with AS-ODN of different concentrations (P < 0.01). The invasiveness inhibition rates were 34.0%, 57.8% and 79.7% at the cells treated with AS-DON of the final concentrations of 0.1 micro mol/L, 0.2 micro mol/L, and 0.4 micro mol/L, respectively.


CONCLUSIONS
Heparanase AS-DON complementary to the start codon region of heparanase mRNA has a significant inhibitory effect on the invasiveness of human mammary carcinoma cell line in a dose-dependent manner.
Zhang
You-lei
YL
Department of General Surgery, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China.
Fu
Zhi-ren
ZR
Zhang
Jun
J
Wang
Yuan-he
YH
Shen
Qian
Q
D000970:Antineoplastic Agents / Q000494:pharmacology; D001943:Breast Neoplasms / Q000473:pathology*; D018387:Codon, Initiator / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D005966:Glucuronidase / Q000096:biosynthesis / Q000235:genetics*; D006801:Humans; D009361:Neoplasm Invasiveness / Q000517:prevention & control; D020319:Oligodeoxyribonucleotides, Antisense / Q000494:pharmacology*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection; D014407:Tumor Cells, Cultured; D000099175:Heparanase
D000970:Antineoplastic Agents; D018387:Codon, Initiator; D005966:Glucuronidase; D020319:Oligodeoxyribonucleotides, Antisense; D012333:RNA, Messenger; D000099175:Heparanase