地鼠颊粘膜癌变过程中增殖细胞核抗原及核仁组成区表达变化实验研究

The Expression of PCNA and NOR in Carcinogenesis Procession of Hamster Buccal Pouch Mucosa

目的 :探讨癌变过程中增殖细胞核抗原 (PCNA)、核仁组成区 (NOR)的表达变化。方法 :利用地鼠颊粘膜癌模型采用免疫组化LSAB法检测PCNA ,阳性对照为人类炎性增生淋巴结标本 ,阴性对照为PBS代替单抗。采用硝酸银染色法检测NOR。结果 :①正常粘膜AgNOR颗粒主要见于基底层细胞 ,以单一型为主。重度异常增生以聚集型为主。浸润癌以混合型为主。②随恶变进展 ,PCNA表达逐渐增强 ,重度异常增生阳性细胞见于上皮全层。结论 :PCNA与AgNOR表达有相关性 (r =0 6 35 ,P <0 0 0 1) ,但PCNA更为直观 ,清晰 ,实用价值更大。

Objective: To investigate the expression of proliferating cell nuclear antigen (PCNA) and nucleolar organizer regions (NOR) with the carcinogenesis model of hamster buccal pouch mucosa. Methods: 48 Syrain hamsters with 6～8 weeks old and 70～80g weight were selected. The material used for inducing cancer is 0 5% DMBA (7,12 dimethylbenzanthracene) in acetone. 0 5% DMBA was applied in the right buccal pouch of the hamster 3 times a week for 12 weeks. The control received no treatment. The time for collecting specimens was 3, 6, 9 and 12 weeks. The specimens were immediately fixed with 10% formalin, dyed with HE, and then two doctors major in histopathology evaluated with the WHO criterion (1986). The wax of immunohistochemical specimen was cut and placed on the APES pieces of glass, treated with 50℃ for 2 hours and preserved under the indoor temperature. The PCNA was examined with the LSAB technique of immunohistochemistry. The company called Zymed provided the MonAb. The positive control was a sample of human inflammatory hyperplastic lymphoma, and the negative was PBS replacing the MonAb. NOR count was determined in a method of silver nitrate staining. The test condition should be at 45℃ without light. Results: 1.AgNOR was usually located in the basal cell layer of normal epithelium, mainly in a single form. High dysplasia was mainly in an aggregated form, however, carcinoma in infiltration appeared to be in a mixed form. 2. PCNA expression was similar to that of the normal control in hyperplastic epithelia (3 weeks) at a moderate level in dysplasia epithelia (6～9 weeks). PCNA expressed highly in squamous cell carcinoma. The expression of the PCNA in line with malignant progress became stronger increasingly. The positive cell was perceivable in all epithelia of high dysplasia. Conclusion: PCNA has been significantly positive and correlative with AgNOR (r=0 635, P<0 001), however, the film of PCNA is much clearer and the practical value is greater.