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Archive > Volume 29 Issue 6 > 2022,29(6):549-556. DOI:10.3872/j.issn.1007-385X.2022.06.006 Prev Next
Clinical Research
Expression of integrin-lined kinase in esophageal squamous cell carcinoma tissues and its effect on proliferation and apoptosis of KYSE-150 cells and the growth of xenografts in nude mice
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Abstract:
Objective: To analyze the expression level of integrin-linked kinase (ILK) gene in esophageal squamous cell carcinoma (ESCC) tissues and its relationship with the clinicopathological characteristics of patients, and to explore its effect on KYSE-150 cell proliferation, apoptosis and the growth of subcutaneous xenograft tumor in nude mice. Methods: The cancer tissues and paired paracancerous tissues of 75 patients with ESCC who had surgical resection and confirmed by pathological examination from January 2012 to December 2014 were selected. Tissue chip technology and immunohistochemical staining were used to detect the expression of ILK in ESCC tissues and para-cancerous tissues; qPCR was used to detect the ILK expression in ESCC cell ECA109, TE-1, EC9706 and KYSE-150, and the KYSE-150 cells with the highest ILK expression was selected for subsequent cell functional studies. The KYSE-150 cells were transfected with ILK interference lentivirus to down-regulate the expression of ILK, and the knockdown efficiency was verified by qPCR and WB methods; the effects of interfering ILK expression on the proliferation and apoptosis of KYSE-150 cells were detected by MTT assay, clone formation assay and FACS; subcutaneous tumorigenesis assay in nude mice was used to detect the effect of interfering ILK on the growth of KYSE-150 cells transplanted tumors. Results: The positive rate of ILK protein in ESCC tissues was higher than that in para-cancerous tissues (P<0.05), and the high expression of ILK was associated with lymph node metastasis (P<0.05). The mRNA expression of ILK in shILK-infected KYSE-150 cells was significantly inhibited (P<0.05)and the expression of ILK protein was significantly down-regulated in the shILK group, which indicated the successful knockdown of ILK. Compared with the cells transfected with shCtrl, the proliferation ability and colony formation number of KYSE-150 cells in the shILK group were significantly reduced (both P<0.05), but the apoptosis rate was significantly increased (P<0.05). Compared with the NC group, the growth of transplanted tumors in the nude mice of the KD group was slower, and the weight and volume of the tumor were smaller (both P<0.05). Conclusion: The expression of ILK in ESCC tissues is higher than that in para-cancerous tissues, and the high ILK expression is associated with lymph node metastasis. Silencing ILK gene can inhibit the proliferation but promote the apoptosis of KYSE-150 cells and inhibit tumorigenesis in nude mice.
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