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目的检测肺鳞癌组织、鳞状细胞不典型增生组织和癌旁组织中磷酸化信号传导及转录激活子3(p STAT3)蛋白的表达,比较其表达与吸烟的关系;探讨STAT3通路在烟草诱导细胞恶性转化过程中的作用。方法免疫组化法检测288例肺鳞癌组织、108例鳞状细胞不典型增生组织、112例癌旁组织中p STAT3蛋白的表达,比较其表达与吸烟的相关性。构建卷烟烟气凝集物(CSC)诱导第10,20,30,40,50,60及70代细胞(P10,P20,P30,P40,P50,P60及P70)永生化人支气管上皮细胞(BEP2D)恶性转化模型。从血清抗性及锚着独立性等方面对CSC诱导各代BEP2D细胞的恶性转化特征进行鉴定。Western蛋白印迹法检测CSC诱导各代BEP2D细胞中p STAT3的表达。MTT法和流式细胞术分别检测JSI-124 0.25~10μmol·L~(-1)对P70细胞存活及凋亡的影响。JSI-124处理CSC诱导P70 24 h后检测存活蛋白表达的变化。结果 p STAT3蛋白在肺鳞癌组织中表达高于鳞状细胞不典型增生和癌旁组织(P<0.05);p STAT3在目前吸烟者中的表达均高于曾经吸烟者和从不吸烟者(P<0.05),且随着吸烟指数增加两者表达水平逐渐升高(P<0.01)。随着转化代数的增高,细胞血清抗性增加,锚着独立性增强,P30细胞以后尤为明显。p STAT3在正常对照组和乙醇对照组中弱表达,且两者之间无显著性差异;CSC诱导的各组BEP2D细胞中,p STAT3的表达均显著高于正常对照组和乙醇对照组(P<0.05),并随细胞代数的增高而增高。JSI-124抑制P70细胞增殖、促进P70细胞凋亡呈浓度及时间依赖性。JSI-124 1μmol·L~(-1)作用P70细胞24 h后,存活蛋白表达显著降低(P<0.05)。结论 STAT3信号通过调控存活蛋白表达抑制细胞凋亡,参与烟草诱导永生化人支气管上皮细胞的恶性转化。
Objective To detect the expression of phospho-signal transducers and activators of transcription 3 (p STAT3) in lung squamous cell carcinoma, squamous cell dysplasia and paracancerous tissues, and to compare the relationship between the expression of p STAT3 and smoking. To explore the role of STAT3 pathway in tobacco induction The role of cell malignant transformation process. Methods Immunohistochemistry was used to detect the expression of p STAT3 protein in 288 cases of squamous cell carcinoma, 108 cases of squamous cell dysplasia and 112 cases of paracancerous tissues. The correlation between the expression of p STAT3 protein and smoking was compared. Cigarette smoke agglutination (CSC) was induced to induce immortalized human bronchial epithelial cells (BEP2D) on the 10th, 20th, 30th, 40th, 50th, 60th, and 70th passages (P10, P20, P30, P40, P50, P60 and P70) Malignant transformation model. The characteristics of CSC-induced malignant transformation of BEP2D cells were identified from aspects of serum resistance and anchorage independence. Western blotting was used to detect the expression of p STAT3 induced by CSC in BEP2D cells. The effects of JSI-124 0.25 ~ 10μmol·L ~ (-1) on the survival and apoptosis of P70 cells were detected by MTT assay and flow cytometry respectively. JSI-124 treatment of CSC-induced P70 24 h after the change in the expression of survivin. Results The expression of p STAT3 protein in squamous cell carcinoma was higher than that in squamous cell atypical hyperplasia and paracancer tissues (P <0.05). The expression of p STAT3 in current smokers was higher than that in smokers and never smokers P <0.05). With the increase of smoking index, the expression levels of both groups increased gradually (P <0.01). With the increase of transformation algebra, the cell serum resistance increased, and the anchorage independence increased, especially after P30 cells. p STAT3 was weakly expressed in normal control group and ethanol control group, and there was no significant difference between the two groups. The expression of p STAT3 in BEP2D cells induced by CSC was significantly higher than that in normal control group and ethanol control group <0.05), and increased with the increase of cell algebra. JSI-124 inhibited the proliferation of P70 cells and promoted the apoptosis of P70 cells in a concentration and time-dependent manner. Survival protein expression of P70 cells treated with 1μmol·L -1 JSI-124 for 24 h was significantly decreased (P <0.05). Conclusion STAT3 signaling inhibits apoptosis through regulating survivin expression and is involved in the malignant transformation of immortalized human bronchial epithelial cells induced by tobacco.