鱼藤酮通过调控JAK/STAT3通路对口腔鳞状细胞癌细胞增殖、凋亡的影响

Effects of rotenone on the proliferation and apoptosis of oral squamous cell carcinoma cells by regulating the JAK/STAT3 pathway

目的:探究鱼藤酮通过调控酪氨酸激酶/信号转导和转录活化因子3(JAK/STAT3)通路对口腔鳞状细胞癌细胞增殖、凋亡的影响。方法:将口腔鳞状细胞癌SCC25细胞分为对照组、低浓度鱼藤酮组(5μmol/L)、中浓度鱼藤酮组(10μmol/L)、高浓度鱼藤酮组(20μmol/L)、JAK2抑制剂AG490组(40μmol/L AG490)。MTT法检测各组SCC25细胞活力;克隆形成实验检测SCC25细胞增殖情况;流式细胞术检测SCC25细胞凋亡情况;Western blotting检测SCC25细胞中增殖、凋亡及通路相关蛋白表达情况。结果:与对照组比较,低浓度鱼藤酮组、中浓度鱼藤酮组、高浓度鱼藤酮组SCC25细胞存活率、克隆细胞数、c-Myc、CyclinD1、p-JAK2/JAK2、p-STAT3/STAT3表达水平呈剂量依赖性显著降低,而凋亡率、caspase-3、Bax表达水平呈剂量依赖性显著增加(P<0.05~P<0.01),AG490组SCC25细胞存活率、克隆细胞数、c-Myc、CyclinD1、p-JAK2/JAK2、p-STAT3/STAT3表达水平显著降低,而凋亡率、caspase-3、Bax表达水平显著增加(P<0.05~P<0.01);与高浓度鱼藤酮组相比,AG490组细胞存活率、克隆细胞数、c-Myc、CyclinD1、p-JAK2/JAK2、p-STAT3/STAT3、凋亡率、caspase-3、Bax表达水平差异均无统计学意义(P>0.05)。结论:鱼藤酮对SCC25细胞增殖的抑制及凋亡的促进可能与抑制JAK/STAT3信号通路有关。

Objective:To explore the effects of rotenone on the proliferation and apoptosis of oral squamous cell carcinoma cells by regulating the Janus kinase/signal transducers and activators of transcription 3(JAK/STAT3)pathway.Methods:Oral squamous cell carcinoma SCC25 cells were divided into control group,low-concentration group(5μmol/L rotenone),medium-concentration group(10μmol/L rotenone),high-concentration group(20μmol/L rotenone),and AG490 group(40μmol/L JAK2 inhibitor AG490).MTT method was used to detect viability of SCC25 cells.Clone formation test was used to detect the proliferation of SCC25 cells.Flow cytometry was used to detect apoptosis of SCC25 cells.Western blotting was used to detect the expression of proliferation,apoptosis and pathway-related proteins in SCC25 cells.Results:Compared with the control group,the survival rate of SCC25 cells,the number of cloned cells,and the expression levels of c-Myc,CyclinD1,p-JAK2/JAK2,and p-STAT3/STAT3 in the low concentration group,medium concentration group,high concentration group decreased significantly in a dose-dependent manner,while the apoptotic rate,caspase-3 and Bax expression levels significantly increased in a dose-dependent manner(P<0.05 to P<0.01);the survival rate of SCC25 cells,number of cloned cells,and the expression levels of c-Myc,CyclinD1,p-JAK2/JAK2,and p-STAT3/STAT3 in the AG490 group decreased significantly,while the apoptotic rate,caspase-3 and Bax expression levels increased significantly(P<0.05 to P<0.01).Compared with the high concentration group,there was no significant difference in cell survival rate,number of cloned cells,c-Myc,CyclinD1,p-JAK2/JAK2,p-STAT3/STAT3,apoptosis rate,caspase-3,Bax expression levels in the AG490 group(P>0.05).Conclusions:The inhibition of proliferation and the promotion of apoptosis of SCC25 cells by rotenone may be related to the inhibition of JAK/STAT3 signaling pathway.