Ras蛋白表达、基因突变在皮肤病理性瘢痕和瘢痕癌中的意义

发布时间：2018-11-20 08:05

【摘要】：目的：探讨Ras基因家族(H-ras/N-ras/K-ras)蛋白表达、基因突变在皮肤病理性瘢痕和瘢痕癌中的意义。方法：以16例皮肤病理性瘢痕、20例皮肤瘢痕癌组织为研究对象,以10例正常皮肤组织为对照。采用免疫组织化学(SP)法分别检测H-ras、N-ras、K-ras蛋白的表达,结合图像分析,分别观测三组被检组织中所检各项指标的表达(平均光密度和阳性面积)。另外对所选病例的石蜡包埋组织(包括14例瘢痕,14例瘢痕癌)提取DNA,进一步做PCR扩增及测序,分析H-ras、N-ras、K-ras基因的第12、13位密码子点突变情况。所有数据运用SPSS16.0软件包进行统计学分析。结果：(1) H-ras、N-ras、K-ras蛋白在正常皮肤表皮和皮肤病理性瘢痕上皮中呈阴性或弱阳性表达,正常皮肤组与瘢痕组比较,差异均无统计学意义(P0.05),但瘢痕上皮中阳性表达细胞数量较正常皮肤表皮增多。(2) H-ras、N-ras、K-ras蛋白在瘢痕癌组织中呈强阳性表达,瘢痕癌组的表达(平均光密度和阳性面积)与正常皮肤组及皮肤瘢痕组比较,差异均有统计学意义(P0.05)。(3)在瘢痕癌中,不管是组织学类型还是细胞的分化,均显示分化越高Ras蛋白表达越强,分化越低Ras蛋白表达越弱,高分化组与低分化组比较,差异均有统计学意义(P0.05)。(4)K-ras阳性信号在正常皮肤组主要定位于细胞核,在瘢痕组主要定位于细胞核和细胞质,在瘢痕癌组为细胞质的表达,出现自胞核向胞质逐渐移位的现象。(5)Ras基因家族点突变分析：从石蜡包埋组织(28例样本)中提取DNA,并经PCR反应后,均成功扩增出目的片段。经测序后,未发现H-ras、N-ras、K-ras基因第12、13位密码子的点突变。结论：(1)在皮肤病理性瘢痕上皮中H-ras、N-ras、K-ras蛋白限制性的低表达,可能与皮肤瘢痕上皮增生具有一定的相关性。(2)瘢痕癌与正常皮肤和皮肤病理性瘢痕相比较,H-ras、N-ras、K-ras蛋白的高表达,可能与皮肤瘢痕癌的发生有关。(3)H-ras、N-ras、K-ras蛋白与肿瘤细胞的分化有相关性,可能在瘢痕癌演进过程中逐渐丢失。(4)K-ras蛋白的移位现象,可能与瘢痕癌的发生有相关性。(5)瘢痕癌中尚未发现Ras基因家族12、13位密码子的突变,其突变位点有待进一步探讨。
[Abstract]:Objective: to investigate the significance of Ras gene family (H-ras/N-ras/K-ras) protein expression and gene mutation in pathological scar and scar carcinoma of skin. Methods: 16 cases of skin pathological scar, 20 cases of skin scar carcinoma and 10 cases of normal skin tissue were studied. Immunohistochemical (SP) method was used to detect the expression of H-rasn- N-rasna-K-ras protein. Combined with image analysis, the expression of each index (average optical density and positive area) was observed in the three groups. In addition, DNA, was extracted from paraffin-embedded tissues (including 14 scars and 14 scar carcinomas) from selected cases. Further PCR amplification and sequencing were performed to analyze the point mutation at codon 1213 of H-rasn N-rascino K-ras gene. All the data were analyzed by SPSS16.0 software package. Results: (1) the expression of H-rasln N-rasna-K-ras protein was negative or weakly positive in normal skin epidermis and skin pathological scar epithelium, but there was no significant difference between normal skin group and scar group (P0.05). But the number of positive cells in scar epithelium was higher than that in normal skin epidermis. The expression (mean optical density and positive area) of scar carcinoma group was significantly different from that of normal skin group and skin scar group (P0.05). (3), whether histological type or cell differentiation. The higher the differentiation, the stronger the expression of Ras protein, the lower the expression of Ras protein. The difference was statistically significant (P0.05). (4) K-ras positive signal was mainly located in nucleus in normal skin group, mainly in nucleus and cytoplasm in scar group, and in cytoplasm in scar carcinoma group. (5) Ras gene family point mutation analysis: DNA, was extracted from paraffin embedded tissue (28 samples) and amplified successfully after PCR reaction. After sequencing, there was no point mutation at codon 1213 of H-rasln N-rasn K-ras gene. Conclusion: (1) the low expression of H-rasln N-rascino K-ras protein in the skin pathological scar epithelium. (2) compared with normal skin and skin pathological scar, the expression of H-rasN rasn K-ras protein in scar carcinoma is higher than that in normal skin and skin pathological scar. It may be related to the development of skin scar carcinoma. (3) H-rasN-rascinia K-ras protein is associated with the differentiation of tumor cells, and may be lost gradually during the progression of scar carcinoma. (4) the translocation of K-ras protein. (5) the mutation of Ras gene family at codon 1213 has not been found in scar carcinomas, and its mutation site needs to be further studied.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R739.5

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