[Inhibitory effects of antisense focal adhesion kinase oligodeoxynucleotides on the invasion of Bel 7402 hepatocellular carcinoma cells].
OBJECTIVE
To study the inhibitory effects of antisense focal adhesion kinase (FAK) oligodeoxynucleotides (ODN) on the invasion of Bel 7402 cells, and investigate the mechanisms.


METHODS
LipofecTAMINE-mediated antisense FAK ODN was transfected into Bel 7402 cells. Cell number and viability were evaluated every 24 hours by trypan blue dye exclusion. Cell attachment assay was carried out at intended time points in a microculture well pre-coated with fibronectin (FN). The invasive activity of tumor cells was assayed in a transwell cell culture chamber. Cell cycle and cell apoptosis analysis were performed with flow cytometry (FCM).


RESULTS
The expression of p125FAK in the group treated with antisense FAK ODN (6.49%+/-0.10%) significantly decreased, compared with those in the group treated with sense FAK ODN (14.33%+/-1.88%) and control group (16.68%+/-1.62%), F=7.66, P<0.01. Antisense FAK ODN significantly inhibited the growth of Bel 7402 cells by 30%-60%, the attachment by 25%-55%, and the invasion, 15%-25%. The decreased expression of FAK in Bel 7402 cells caused a G2/M cell cycle arrest, and the cells at S phase decreased significantly. The occurrence of apoptosis detected by FCM increased significantly in the group treated with antisense FAK ODN.


CONCLUSIONS
Inhibition of FAK expression significantly decreases the attachment between ECM and Bel 7402 cells, and the ability of Bel 7402 cells to invade the reconstituted basement membrane. In addition, FAK suppression significantly inhibits the proliferation of Bel 7402 cells in vitro, and increases their apoptosis.
Department of Surgery, Ninth People's Hospital, Shanghai Second Medical University, Shanghai 200011, China.
Yan
Y
Gu
Ji-sheng
JS
Chen
Xiao-dong
XD
Zhou
D017209:Apoptosis / Q000187:drug effects; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D002448:Cell Adhesion; D002455:Cell Division / Q000235:genetics; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; D015971:Gene Expression Regulation, Enzymologic; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D009361:Neoplasm Invasiveness; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D011505:Protein-Tyrosine Kinases / Q000494:pharmacology*; D014162:Transfection
D016376:Oligonucleotides, Antisense; D011505:Protein-Tyrosine Kinases; D051417:Focal Adhesion Kinase 1; D051416:Focal Adhesion Protein-Tyrosine Kinases; C496050:PTK2 protein, human