| The interaction of cell and ECM ( extracellular matrix ) is involved multiple processes of cell acticity in growth , survival , differentiation, proliferation and migration. Apoptosis would be starteded if cell lost the balance of this interaction. Recently, studies revealed that decreased interaction of cell and ECM has close relationship with the process of differentiation, proliferation and migration of tumor cells. Integrin plays an important role in extracellular signal transduction and mediates adhesion of cells to ECM. ILK(integin-linked kinase), which is a critical component in the integrin signaling pathway, controls many downstream targets that connect integrin to its effectors in vivo. For ILK regulates multiple cell functions, minim change in activity of ILK protein or in gene expression of ILK would possibly result in the formation and development of tumors. Therefore, ILK may be a promising therapeutic target for tumors.Objective: Using ASODN technique, we investigated the effects of ILK ASODN on the human gastric cancer cell line SGC-7901 by analyzed theexpression changes of ILK mRNA and ILK protein, we want to give further evidence that ILK might be a promising therapeutic target for tumors.Methods:. ( 1 ) The Antisense Oligonucletide of ILK ASODN was transferred into human gastric cancer cell line SGC-7901 by lipofectamine 2000 after it designed and synthesized. Then, we analyzed the changes of ILK level bom in mRNA and in protein by RT-PCR and Western blot in respectivly. (2) The rates of growth and apoptosis in gastric cell line SGC-7901 was measured by MTT assay and FCMS.Results: (1) At 48h of exposure to ASODN, there were two 984bp and 584bp straps by electrophoresis after PCR. By semi-quantity analysis of gel image, the results showed that expression of ILK mRNA has no significant difference in different dose of ASODN treatment groups and do so when compared with control, single transfected Lipofectamine2000 group in respectivly. Interestingly, SODN groups, which showed an obvious decrease of ILK mRNA expression, has significant difference compared with other groups. (2) By analyzing data from Western Blot, it suggested that ILK ASODN could specifically block the expression of ILK protein in gastric cell line SGC-7901 by concentration-dependent manner. In addition, the SODN group also showed inhibitory effect on expression of ILK protein. (3) Cell proliferation in gastric cell line SGC-7901 was inhibited effectively by ILK ASODN and this effect depended on the concentration of incubating solution. After treatment of ILK ASODN at 4umol / L for 24h, the largest inhibitioncould be achieved compared with 2umol / L and lumol / L, whose largest inhibition is obtained at 48h and 72h in respectivly. (4) After cell was incubated with ILK ASODN (lumol / L, 2umol / L,4umol / L) for 48h, the apoptosis rate were 6.42%, 18.75%, 35.61% in respectivly by FCM measurement, whereas it were 3.85% and 3.72% in groups of single transfected lipofectamine and control in respectively. It indicated that the increase of apoptosis rate dependent on the dose of ASODN treatment.Conclusion: In order to inhibit the gastric cancer proliferation, we designed an ILK ASODN and transferred it into gastric cancer cells to block the expression of ILK protein. Our results show that ILK ASODN could specifically block the expression of ELK protein, inhibit cells proliferation and induce apoptosis, although it has no effect on transcription of ILK mRNA. It suggests that ILK plays an important role hi cell growth, proliferation and apoptosis and might be a promising therapeutic target for tumors.
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