shRNA Interferes with Cystatin C to Inhibit Invasion and Migration of Non-small Cell Lung Cancer A549 Cells

Abstract

Objective To investigate the effect of Cystatin C (Cys C) expression on the growth, invasion and metastasis of NSCLC A549 cells and its potential molecular mechanism. Methods sh-Cys C vector was constructed using short hairpin RNA (sh-RNA) to knock down Cys C expression, and transfected into human lung adenocarcinoma A549 cells. sh-Cys C stably expressional cell line was screened. Cell proliferation was detected by CCK-8. Cell migration was detected by wound healing test. Cell invasion was detected by Transwell assay. In addition, Western blot analysis was used to detect the expression changes of key proteins in invasion and migration. Results shRNA interfered with Cys C and significantly decreased the expression of Cys C. Compared with the control group, shRNA-mediated Cys C silencing significantly inhibited the proliferation, migration and invasion of A549 in vitro. In addition, the downregulation of Cys C could significantly inhibit the expression of matrix metalloproteinase (MMP9, MMP14) and vascular endothelial growth factor (VEGF). Conclusion shRNA interferes with Cys C and significantly down-regulates the expression of Cys C, which could inhibit the invasion and migration of A549 cells.

Keywords

• cystatin c
• migration
• invasion
• non-small cell lung cancer