Effects Of Silencing Cyclophilin A By SiRNA On Proliferation,Metastasis And Chemosensitivity To Cisplatin In Hep-2 Cells

Objective: Laryngeal carcinoma is one of the most common malignancies of human head and neck cancers, and laryngeal squamous cell carcinoma(LSCC) represents approximately 99%. LSCC has an obviously growing trend in recent years. Thus, understanding of the mechanisms of laryngeal carcinogenesis and identification of novel therapeutic targets for LSCC is crucial. Cyclophilin A(Cyp A) is the intracellular receptor for immunosuppressive drug cyclosporin A(Cs A).Cyp A overexpression has been found in a variety of human malignancies,including colon cancer, pancreatic cancer, hepatocellular carcinoma and some other malignant tumors. Our previous studies had found the expression of Cyp A in laryngeal carcinoma tissues was significantly higher than that in normal adjacent tissues, but the biological function and mechanism in LSCC was unclear. This study is to investigate the effect of silencing Cyp A on proliferation and metastasis of Laryngeal carcinoma cell line Hep-2 and its mechanism.Methods: Si RNA targeting Cyp A and negative control si RNA weretransfected into Hep-2 cells. Real-time PCR was used to detect the m RNA expression of Cyp A, CD147, VEGF and MMP-9. Protein expression of Cyp A, CD147, MMP-9 and VEGF was detected by Western blot and ELISA. The impact of Cyp A on cell growth and cisplatin sensitivity was detected by MTT assay. Colony formation assay was used to examine the role of Cyp A in cell proliferation; cell cycle distribution was measured by flow cytometry; Transwell test was used to test the invasion and migration ability of Hep-2 cells; vasculogenic mimicry assay was used to detect the ability of vascular channel formation about Hep-2 cells.Results: Si RNA Cyp A markedly decreased both Cyp A m RNA and protein expression( P=0.000). Knockdown of Cyp A significantly down-regulated the m RNA and protein expression of CD147, VEGF, and MMP-9. MTT assay showed that the proliferation of Cyp A si RNA group was inhibited significantly and the chemosensitivity to cisplatin in Cyp A si RNA group was markedly increased compared with control group and NC si RNA group(P=0.000).Si RNA Cyp A reduced the colony formation,and the number of colonies in Cyp A si RNA group were(129.33±14.22),significantly less than in control( 235.00±23.90) and NC si RNA(240.67±10.07)(P=0.000); cell cycle was arrested in G1 phase and the cell proliferation index of each group was control(0.337±0.016)、 NC si RNA(0.345±0.024)、Cyp A si RNA(0.191±0.005)(P=0.000); the migration, invasion and vasculogenic mimicry ability of Hep-2 cellswas distinctly decreased(P=0.000).Conclusion: Down-regulation of Cyp A can inhibit the proliferation,migration, invasion and VM ability, and increase the sensitivity to cisplatin of Hep-2 cells, which may be related to the decreased of CD147, VEGF and MMP-9.