sigma-1受体对口腔鳞癌细胞增殖、迁移和侵袭影响的实验研究

Experimental study on the effect of sigma-1 receptor on proliferation,migration and invasion of oral squamous cell carcinoma

目的探究sigma-1受体在口腔鳞状细胞癌(OSCC)组织和细胞系中的表达,并通过特异性siRNA转染降低sigma-1受体在OSCC细胞中的表达,检测对其生物学行为的影响,并初步分析其可能的机制。方法免疫组化法检测人OSCC组织中sigma-1受体的表达。Western blot和免疫荧光法检测人OSCC细胞系中sigma-1受体的表达。利用GEPIA数据库分析sigma-1受体表达水平与OSCC患者预后的相关性。通过转染特异性siRNA下调sigma-1受体表达,CCK-8和Transwell迁移侵袭实验检测转染后OSCC细胞的增殖、迁移和侵袭,qRT-PCR和Western blot检测转染后电压门控钠离子通道亚型Nav1.5在OSCC细胞中的表达水平。结果免疫组化分析结果显示sigma-1受体在OSCC组织中较正常组织为高表达。Western blot检测结果显示,sigma-1受体在4种OSCC细胞系中均有表达。GEPIA数据库分析结果显示,sigma-1受体的表达水平与OSCC患者的总体生存率呈负相关。通过细胞免疫荧光实验结果显示sigma-1受体在CAL-27细胞中主要定位在质膜上。CCK-8分析结果显示sigma-1受体通过siRNA下调表达后降低了CAL-27细胞的增殖。Transwell分析结果显示降低sigma-1受体表达可显著抑制CAL-27细胞的迁移和侵袭。此外,下调sigma-1受体表达后Nav1.5的表达水平被抑制。结论sigma-1受体参与OSCC的发生发展,促进了OSCC细胞的增殖、迁移和侵袭,并可能通过调节Nav1.5离子通道参与OSCC的进展。

Objective To investigate the expression of sigma-1 receptor in oral squamous cell carcinoma(OSCC)tissues and cell lines,reduce the expression of sigma-1 receptor in OSCC cells by transfection with specific siRNA,detect the effect on their biological behavior,and preliminarily analyze the possible mechanism.Methods The expression of sigma-1 receptor in human OSCC tissues were detected by immunohistochemistry.Western blot and immunofluorescence were used to detect the expression of sigma-1 receptor in human OSCC cell lines.GEPIA database was used to analyze the correlation between sigma-1 receptor expression level and prognosis of patients with OSCC. The expression of sigma-1 receptor was down-regulated by transfection of specific siRNA. Then the proliferation, migration and invasion of OSCC cells after transfection were detected by CCK-8 and Transwell assay, and the expression level of voltage-gated sodium ion channels subtype Nav1.5 in OSCC cells was detected by qRT-PCR and Western blot. Results Immunohistochemical analysis showed that the expression of sigma-1 receptor was higher in OSCC tissues than that in normal tissues. Western blot revealed that the sigma-1 receptor was expressed in all four OSCC cell lines. The results of GEPIA database analysis showed that the expression level of sigma-1 receptor was negatively correlated with the overall survival rate of patients with OSCC. Immunocytochemistry and confocal microscopy showed that the sigma-1 receptor was mainly localized in the plasma membrane in CAL-27 cells. CCK-8 analysis showed that the down-regulated expression of sigma-1 receptor reduced the proliferation of CAL-27 cells through its specific siRNA. Transwell analysis showed that the reduced expression of sigma-1 receptor significantly inhibited the migration and invasion of CAL-27 cells. In addition, down-regulation of sigma-1 receptor expression was found to inhibit the protein and mRNA expression levels of Nav1.5. Conclusion sigma-1 receptor participates in the occurrence and development of OSCC, promotes the proliferation, migration and invasion of OSCC cells, and may be involved in the progression of OSCC by regulating the Nav1.5.