Effects Of Antisense Oligonucleotide Targeting Integrin-Linked Kinase On Epithelial-Mesenchymal Transition And Laryngeal Carcinoma Metastasis

Background and Aims Laryngeal squamous cell carcinoma(LSCC) is one of the most frequently diagnosed malignancies of head and neck in the world. LSCC recurrence and metastasis lead to the failure of treatment. Multi-genes and multi-factors are involved in the processes of LSCC recurrence and metastasis. Elucidating molecular mechanisms of LSCC recurrence/metastasis and searching for effective target therapies are the promising pathway to improve the survival.Integrin-Linked Kinase(ILK) is a 59KD cytoplasmic signaling protein, a serine/threonine protein kinase. Evidence now points to ILK as having crucial, multifaceted roles in the normal development and function of several tissues. ILK plays a central role in transducing many of the biochemical signals that are initiated by cell-matrix interactions and that regulate anchorage-dependent cell growth and survival, cell-cycle progression, epithelial mesenchymal transition(EMT), invasion and migration motility, vascular development, and tumour angiogenesis.Recently, epithelial-mesenchymal transitions (EMT) and mesenchymal to epithelial transition (MET), the fundamental processes regulating morphogenesis in multicellular organisms, is thought to play a key role during early steps of invasion and metastasis of epithelial malignancies. EMT involves an orchestrated series of events in which epithelial cells lose polarity and cell-cell contacts and undergo dramatic remodeling of the cytoskeleton which enables carcinoma cells to migrate through an extracellular environment and settle in distinct areas to form metastasis.As a EMT inducer, ILK is thought to play a key role in regulating developmental EMT and tumor metastasis. The roles and mechanisms of ILK in human laryngeal squamous cell carcinoma progression has not yet been fully characterized and has not been reported to date. Taken this, the present study is designed to investigate the effect of ILK on cells migration and invasion in laryngeal squamous cell carcinoma.Method The expression of ILK protein relation to clinicopathologic characteristics in 56 cases laryngeal squamous cell carcinoma were detected by immunohistochemical methods. Lipofectamine-mediated transfection was used to introduce a ILK antisence oligonucleotide into laryngeal squamous cell carcinoma Hep-2 cell lines.Transfectants were tested in invasion and migration assay by a Transwell chamber and the expression of epithelial-mesenchymal transition associated genes by western blot analysis.Results The expression of ILK protein positive rate is significantly difference between 56 cases laryngeal squamous cell carcinoma(LSCC) tissues 66.07% and 28 cases non-carcinoma laryngeal tissues (NCLT) 21.43%(P＜0.05). ILK protein expression were significantly correlated with LSCC pathological grade and the presence of lymph node metastasis (P<0.05). overexpression of ILK in laryngeal squamous cell carcinoma Hep-2 cell lines was significantly suppressed by ILK antisence Oligonucleotide(ASOs). ILK ASOs inhibit Hep-2 cell lines growth and promote cell apoptosis. Mechanisms studies revealed that suppression of ILK enhanced the recruitment of E-cadherin and had a concomitant effect of down-regulated expression of Vimentin, which, in turn, lead to significantly decreased adhesive, migrant and invasive abilities in Hep-2 cell lines.Conclusion Taken together, our results indicate the importance of ILK as an invasion-promoting gene in Hep-2 cell lines through EMT introduction.