体外siRNA-Nav1.5干扰口腔鳞癌HSC-3细胞生物学行为研究

siRNA-Nav1.5 interferes HSC-3 cells and changes in biological behavior in vitro

目的 探究电压门控钠离子通道(VGSC)亚型Nav1.5在口腔鳞癌细胞系和组织中的表达及小干扰RNA(siRNA)转染对HSC-3口腔鳞癌细胞的生物学行为的影响,并进一步分析其可能机制。方法 选取临床口腔鳞癌组织和正常口腔上皮组织免疫组化染色显示Nav1.5在组织中的表达。免疫荧光激光共聚焦显示细胞内Nav1.5的表达。利用脂质体转染siRNA-Nav1.5干扰HSC-3细胞中Nav1.5的表达水平,CCK8和Transwell侵袭实验检测转染后细胞的增殖和侵袭能力。实时荧光定量PCR(RT-PCR)和Western blot验证与细胞增殖侵袭相关的蛋白:细胞增殖核抗原(PCNA)、基质金属蛋白酶2(MMP-2)、MMP-9表达变化。结果 免疫组化结果可见在口腔鳞癌组织中Nav1.5阳性表达,正常组织中未见明显表达。免疫荧光结果可见HSC-3细胞中Nav1.5阳性表达。转染siRNA-Nav1.5明显降低HSC-3细胞的增殖和侵袭能力。转染siRNA-Nav1.5的HSC-3细胞PCNA、MMP-2和MMP-9蛋白表达降低。结论 Nav1.5可通过调控PCNA、MMP-2和MMP-9的表达促进口腔鳞癌细胞的增殖和侵袭能力。

Objective To study the expression and biological effects of voltage-gated sodium channel 1.5 subtype(Nav1.5)on oral squamous cell carcinoma cell lines and tissues transfected with small interfering RNA(siRNA-Nav1.5),and analyze its possible regulatory mechanism.Methods Oral squamous cell carcinoma tissues and normal oral epithelium tissues were selected from clinical,immunohistochemistry was used to detect the expression of Nav1.5.Confocal microscope showed intracellular expression of Nav1.5.The expression levels of Nav1.5 in HSC-3 cells were transfected with siRNA-Nav1.5 lipidosome.Transwell and CCK8 were used to detect the invasion and proliferation of HSC-3 cells.Western blot and RT-PCR were used to validate the expression of related protein PCNA,MMP-2 and MMP-9.Results Immunohistochemical results showed that Nav1.5 was positively expressed in oral squamous cell carcinoma,but no significant expression was observed in normal tissues.Immunofluorescence results showed positive expression of Nav1.5 in HSC-3 cells.The invasion and proliferation of HSC-3 cells transfected with siRNA-Nav1.5 were significantly suppressed.The expression of PCNA,MMP-2 and MMP-9 protein was decreased in HSC-3 cells after transfected with siRNA-Nav1.5.Conclusion Nav1.5 can promote the proliferation and invasion of oral squamous cell carcinoma by regulating the expression of PCNA,MMP-2 and MMP-9.