Therapeutic Effects Of Tyroservatide On Metastasis Of Lung Cancer And Its Mechanism By Affecting Integrin-FAK Signal Transduction

Objective:To investigate the therapeutic effects of tripeptide tyroservatide (YSV) on metastasis of human lung cancer and explore its possible mechanism by affecting integrin-focal adhension kinase (FAK) signal transduction in tumor cells.Methods:1. MTS assay was applied for adhension to Matrigel of 95D, A549 and NCI-H1299 human lung cancer cells with high metastatic capability by YSV in vitro.2. Transwell model was applied for invasion of 95D, A549 and NCI-H1299 human lung cancer cells by YSV in vitro.3. Gelatin zymography assay was applied for MMP-2 and MMP-9 activities from supernatant of 95D, A549 and NCI-H1299 human lung cancer cells by YSV in vitro; Western blot was applied for matrix metalloproteinases (MMPs) MMP-2 and MMP-9 protein expression of three human lung cancer cells by YSV in vitro; and Real-time PCR was applied for MMP-2 and MMP-9 mRNA level of three human lung cancer cells by YSV in vitro.4. Western blot was applied for FAK Tyr397和FAK Tyr576/577 phosphorylation of 95D, A549 and NCI-H1299 human lung cancer cells by YSV in vitro; Western blot was applied for FAK protein expression of three human lung cancer cells by YSV in vitro; and Real-time PCR was applied for FAK mRNA level of three human lung cancer cells by YSV in vitro.5. Flow cytometry was applied for integrinβ1 and integrinβ3 protein expression of 95D, A549 and NCI-H1299 human lung cancer cells by YSV in vitro; and Real-time PCR was applied for integrinβ1 and integrinβ3 mRNA level of three human lung cancer cells by YSV in vitro.Results:1. YSV could significantly inhibited the adhension to Matrigel of 95D, A549 and NCI-H1299 human lung cancer cells with high metastatic capability by YSV in vitro, with the highest inhibition ratio 37.08%,36.87% and 41.34%.2. YSV could significantly inhibited the invasion of 95D, A549 and NCI-H1299 human lung cancer cells by YSV in vitro, with the highest inhibition ratio 53.87%, 62.10% and 64.44%.3. YSV could significantly inhibited MMP-2 and MMP-9 activities from supernatant of 95D, A549 and NCI-H1299 human lung cancer cells by YSV in vitro; YSV could significantly inhibited MMP-2 and MMP-9 mRNA level and protein expression of 95D, A549 and NCI-H1299 human lung cancer cells by YSV in vitro.4. YSV could significantly inhibited FAK Tyr397 and FAK Tyr576/577 phosphorylation of 95D, A549 and NCI-H1299 human lung cancer cells by YSV in vitro; YSV could significantly inhibited FAK mRNA level and protein expression of 95D, A549 and NCI-H1299 human lung cancer cells by YSV in vitro.5. YSV could significantly inhibited integrin (31 and integrinβ3 mRNA level and protein expression of 95D, A549 and NCI-H1299 human lung cancer cells by YSV in vitro.Conclusion:YSV could inhibit adhension and invasion of human lung cancer cells and exhibit therapeutic effects on metastasis of lung cancer. The anti-metastatic effects of YSV maybe as follows:YSV down-regulated mRNA level and protein expression of integrinβ1 and integrinβ3, reduced the FAK activity and expression, thus inhibited the integrin-FAK signal transduction, which resulted in the activities and expression of this signal transduction downsteam effector molecules MMP-2 and MMP-9 were degraded.