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目的 探讨p5 3、K ras基因突变、蛋白表达在 3 甲基胆蒽 (3 methylcholanthrene ,MCA)和二乙基亚硝胺 (diethylinitrosamine ,DEN)诱发大鼠肺鳞癌发生演进中的作用 ,及其突变与蛋白表达的关系。方法 将大鼠诱发肺癌石蜡标本连续切片 ,切片用于HE染色确定肺癌发生发展的病变阶段 ,及免疫组织化学 (SP法 )检测各阶段p5 3、K ras蛋白表达 ,并用于显微切割 ,定点对位分别切割由正常支气管黏膜上皮细胞演变成癌细胞 ,癌浸润、转移各阶段病灶的主质 ,提取DNA ,用聚合酶链反应 单链构象多态性 (PCR SSCP)检测各阶段p5 3、K ras基因的突变。结果 30例正常支气管黏膜上皮未检测到p5 3、K ras基因突变及其蛋白表达。在 32例支气管黏膜增生和鳞状化生、2 1例不典型增生、12例原位癌、4 3例浸润癌及 17例转移癌组织中 ,p5 3基因突变率分别为 3 1% ,2 8 6 % ,30 0 % ,5 1 2 % ,5 2 9% ;p5 3蛋白阳性表达率分别为 0 ,4 2 9% ,5 0 0 % ,6 0 5 % ,6 4 7% ;不典型增生阶段与增生、鳞状化生阶段相比 ,p5 3基因突变率及蛋白表达率增高 ,差异均有显著性意义 (P <0 0 2 5 ,P <0 0 0 5 ) ,p5 3基因突变及蛋白阳性表达高度相关 (P <0 0 0 5 ,Pearson′sR =0 5 996 )。K ras基因突变率分别为 0 ,4 8% ,8 3% ,9 3
Objective To investigate the role of p5 3, K ras gene mutation and protein expression in the development of lung squamous cell carcinoma in rats induced by 3 methylcholanthrene (MCA) and diethylinitrosamine (DEN) Mutation and protein expression. Methods The paraffin sections from lung cancer were sectioned and stained with HE staining to determine the pathological stage of lung cancer. The expressions of p5 3 and K ras proteins were detected by immunohistochemistry (SP method) and used for microdissection. The normal tissues of bronchial mucosa epithelial cells were evacuated from the normal bronchial mucosa epithelium and the main components of cancer infiltration and metastasis were detected by DNA sequencing. The expression of p53 was detected by polymerase chain reaction single strand conformation polymorphism (PCR SSCP) K ras gene mutation. Results No mutation of p5 3 and K ras gene and its protein expression were detected in 30 cases of normal bronchial epithelium. In 32 cases of bronchial mucosa hyperplasia and squamous metaplasia, 21 cases of atypical hyperplasia, 12 cases of carcinoma in situ, 43 cases of invasive carcinoma and 17 cases of metastatic carcinoma, the mutation rate of p5 3 gene was 31%, 2 The positive rate of p5 3 protein was 0 4 2 9%, 500 0%, 65 0%, 64 7%, respectively; Compared with the stage of hyperplasia and squamous metaplasia, the mutation rate and protein expression of p5 3 gene in proliferative stage were significantly higher than those in proliferative stage (P <0.05, P <0.05) And protein positive expression (P <0 0 0 5, Pearson’s R = 0 5 996). K ras gene mutation rates were 0, 48%, 83%, 93