摘要
目的:观察抑制食管癌斯钙素-1(STC-1)基因表达对食管癌细胞凋亡、IL-1β和TNF-α表达及JAK2/STAT3信号的影响。方法:RT-PCR及Western blot检测STC-1在人食管鳞状细胞癌KYSE170、Eca109、TE1和TE10细胞相对于正常人食管鳞状上皮细胞Het-1A的表达;将合成的STC-1的siRNA序列及无干扰作用的si NRA序列转染Eca109细胞,分别标记为STC-1-siRNA组和NC组,并设定空白对照组,收集转染48 h的细胞,RT-PCR及Western blot检测转染后的细胞中STC-1的表达; CCK8及流式细胞仪分别检测Eca109细胞活力及凋亡率; RT-PCR检测IL-1β和TNF-α表达; Western blot检测Ki67、p53、磷酸化的蛋白酪氨酸激酶2(p-JAK2)、磷酸化的信号转导与转录因子3(p-STAT3)的蛋白表达。结果:与Het-1A细胞比较,STC-1在KYSE170、Eca109、TE1和TE10细胞中的mRNA及蛋白表达均显著升高(P<0. 05);与对照组比较,STC-1-siRNA组STC-1的表达显著降低,细胞活力显著降低,细胞凋亡率显著升高,IL-1β、TNF-α、Ki67、p-JAK2和p-STAT3的表达均显著降低,p53的表达显著升高(P<0. 05)。结论:STC-1在食管癌细胞中高表达,抑制其表达后癌细胞活力显著降低,凋亡率升高,此作用可能与抑制JAK2/STAT3信号通路及炎症因子如IL-1β和TNF-α表达有关。
Objective:To observe the effect of STC-1 gene expression was inhibited on the apoptosis,IL-1βand TNF-αexpression and JAK2/STAT3 signal in esophageal cancer cells.Methods:Compared with normal human esophageal squamous epithelial cells Het-1A,STC-1 expression was detected in human esophageal squamous cell carcinoma KYSE170,Eca109,TE1 and TE10 cells by RT-PCR and Western blot;the siRNA sequence that the synthesized STC-1 and the siNRA sequence without interference were transfected into Eca109 cells,which were labeled as STC-1-siRNA group and NC group,and the blank control group was set,cells were transfected for 48 h,the expression of STC-1 were detected by RT-PCR and Western blot.Eca109 cell viability and apoptosis rate were detected by CCK8 and flow cytometry.IL-1βand TNF-αexpression were detected by RT-PCR;the expression of Ki67,p53,p-JAK2 and p-STAT3 protein were detected by Western blot.Results:Compared with Het-1A cells,expression of STC-1 mRNA and protein in KYSE170,Eca109,TE1 and TE10 cells were increased significantly(P<0.05);compared with the control group,STC-1 expression was decreased significantly in STC-1-siRNA group,cell viability was decreased significantly in STC-1-siRNA group,the apoptosis rate was increased significantly in STC-1-siRNA group;IL-1β,TNF-α,Ki67,p-JAK2 and p-STAT3 expression were decreased significantly in STC-1-siRNA group,p53 expression was increased significantly in STC-1-siRNA group(P<0.05).Conclusion:STC-1 was highly expressed in esophageal cancer cells;inhibiting of STC-1 expression could significantly reduce the activity of cancer cells,and increase apoptosis rate,this effect may be related to the inhibition of JAK2/STAT3 signaling pathways and inflammatory factors as IL-1βand TNF-α.
作者
卜秀梅
王文刚
李辉
郑瑾
BU Xiu-Mei;WANG Wen-Gang;LI Hui;ZHENG Jin(Liaoning University Traditional Chinese Medicine,Shenyang 110032,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2019年第2期186-191,共6页
Chinese Journal of Immunology
