舒芬太尼对人口腔鳞癌细胞系CAL27生长、侵袭和干细胞样特性的影响及线粒体凋亡通路的调节舒芬太尼对人口腔鳞癌细胞系CAL27生长、侵袭和干细胞样特性的影响及线粒体凋亡通路的调节

Effects of sufentanil on the growth,invasion and stem cell-like properties of human oral squamous cell line CAL27 and the regulation of mitochondrial apoptosis pathway

目的:探究舒芬太尼对人口腔鳞癌细胞系CAL27生长、侵袭和干细胞样特性的影响及线粒体凋亡通路的调节。方法:选择不同浓度的舒芬太尼处理人口腔鳞癌CAL27细胞,24 h后CCK-8检测细胞存活率;选择舒芬太尼浓度(0 nmol/L、10 nmol/L、25 nmol/L、50 nmol/L)进行后续实验:BrdU检测细胞增殖;Transwell检测细胞侵袭;蛋白印迹法检测VEGF和Vimentin的蛋白表达;肿瘤细胞成球实验检测肿瘤干细胞增殖;流式分选ALDH阳性细胞;蛋白印迹法检测干细胞标记物和线粒体凋亡通路关键基因的蛋白表达。结果:舒芬太尼浓度不低于25 nmol/L时,CAL27细胞的存活率显著降低,且呈浓度依赖性(P<0.05)。与舒芬太尼0 nmol/L组比较,舒芬太尼25 nmol/L组和舒芬太尼50 nmol/L组BrdU阳性细胞数,侵袭细胞数,肿瘤干细胞成球数均显著减少(P<0.05);VEGF、Vimentin、SOX2、OCT4、c-Myc蛋白表达显著下调(P<0.05);Caspase-3蛋白表达和Bax/Bcl-2比值显著上调(P<0.05);舒芬太尼10 nmol/L组、舒芬太尼25 nmol/L组和舒芬太尼50 nmol/L组ALDH阳性细胞数均明显减少(P<0.05)。结论:舒芬太尼可抑制人口腔鳞癌细胞系CAL27生长、侵袭、降低口腔鳞癌干细胞样特性,促进CAL27细胞凋亡。

Objective:To investigate the effects of sufentanil on the growth,invasion and stem cell-like characteristics of human oral squamous cell line CAL27 and the regulation of mitochondrial apoptosis pathway.Methods:Human oral squamous cell carcinoma CAL27 cells were treated with different concentrations of sufentanil(0 nmol/L,10 nmol/L,25 nmol/L,and 50 nmol/L),and cell survival rate was detected by CCK-8 after 24 h.Cell proliferation was detected by BrdU method.Cell invasion was evaluated by Transwell assay.The expressions of VEGF,Vimentin,stem cell-like markers and key proteins of the mitochondrial apoptotic pathway were determined by Western blotting.The proliferation of cancer stem-like cells was examined by tumor sphere formation assay.ALDH-positive cells were sorted by flow cytometry.Results:When the concentration of sufentanil was higher than 25 nmol/L,the survival rate of CAL27 cells was significantly reduced in a concentration-dependent manner(P<0.05).The number of BrdU-positive cells,invaded cells and tumor spheres,and the expression levels of VEGF,Vimentin,SOX2,OCT4 and c-Myc protein were significantly decreased in 25 nmol/L and 50 nmol/L sufentanil groups,while the Caspase-3 expression level and Bax/Bcl-2 ratio were significantly increased compared with sufentanil 0 nm group(P<0.05).The number of ALDH-positive cells was decreased significantly in 10 nmol/L,25 nmol/L and 50 nmol/L sufentanil groups(P<0.05).Conclusion:Sufentanil can inhibit the growth and invasion of human oral squamous carcinoma cell line CAL27,reduce the stem cell-like characteristics of oral squamous carcinoma,and promote the apoptosis of CAL27 cells.