Cx26、Cx32的表达、基因突变在病理性皮肤瘢痕及瘢痕癌中的意义

发布时间：2018-05-21 01:30

本文选题：Cx26 + Cx32　；参考：《遵义医学院》2011年硕士论文

【摘要】：目的：探讨Cx26、Cx32蛋白和分子水平的表达、基因突变在病理性皮肤瘢痕及瘢痕癌中的意义。方法：以病理性皮肤瘢痕、皮肤瘢痕癌组织为研究对象,以正常皮肤组织为对照。采用免疫组织化学(S-P)法分别检测Cx26、Cx32蛋白的表达,采用核酸分子原位杂交法检测Cx26mRNA、Cx32mRNA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(阳性面积与平均光密度),所有数据输入计算机后运用SPSS16.0软件包进行统计学分析；从14例皮肤瘢痕和14例瘢痕癌石蜡包埋组织中提取DNA,运用PCR扩增及扩增目的片段直接测序的方法,进行突变筛查。结果：(1)Cx26蛋白、Cx26mRNA和Cx32蛋白、Cx32mRNA在瘢痕癌上皮中呈弱阳性或阴性表达。瘢痕癌组分别与正常皮肤组、皮肤瘢痕组比较,表达(阳性面积与平均光密度)差异均有统计学意义(P0.01)；(2)Cx26蛋白、Cx26mRNA在病理性瘢痕上皮中呈强阳性表达,与正常皮肤组比较,其表达(阳性面积与平均光密度)差异有统计学意义(P0.01); (3) Cx32mRNA在病理性瘢痕上皮的表达呈阳性,与瘢痕癌组比较,其表达(阳性面与积平均光密度)差异有统计学意义(P0.01),与正常皮肤组比较,差异无统计学意义(P0.05)。(4)Cx26基因点突变分析：经PCR反应后,在28例患者样本中,扩增出7例目的条带,其中6例瘢痕1例瘢痕癌,经测序分析检测到4例瘢痕的Cx26基因有良性突变,为两种不同类型的多态性碱基变异。(5)Cx32基因点突变分析：经PCR反应后,在28例样本中,扩增出6例目的条带,其中4例瘢痕2例瘢痕癌,经测序分析均未发现致病性与非致病性突变位点。结论：(1)瘢痕癌中Cx26及其mRNA和Cx32及其Cx32mRNA的阴性或弱阳性、低表达,可能与其抑制肿瘤生长功能的丧失,导致瘢痕癌的发生有相关性；(2)瘢痕上皮中Cx26及其mRNA的强阳性、高表达,可能与促进瘢痕上皮增生有相关性,同时也与病理性瘢痕易形成溃疡、溃疡经久不愈有相关性；(3)瘢痕上皮中Cx32mRNA表达水平的降低,有可能是瘢痕癌变的早期事件,值得重视；(4) Cx26、Cx32基因在瘢痕癌的发生发展过程中尚未发现有致病性的基因突变位点,其意义有待进一步探讨。
[Abstract]:Objective: to investigate the expression of Cx26Cx32 protein and its molecular level and the significance of gene mutation in pathological skin scar and scar carcinoma. Methods: pathological skin scar and skin scar carcinoma were studied, and normal skin tissue was used as control. The expression of Cx26mRNA-Cx32 mRNA was detected by immunohistochemistry, and the expression of Cx26mRNA-Cx32 mRNA was detected by in situ hybridization, and the expression of Cx26mRNA-Cx32 mRNA was analyzed by image analysis. The positive area and average optical density were observed in the three groups of tissues. All the data were input into the computer and analyzed by SPSS16.0 software package. DNA was extracted from 14 cases of skin scar and 14 cases of cicatricial carcinoma in paraffin embedded tissue. The mutation was screened by PCR amplification and direct sequencing of the target fragment. Results the expression of Cx26 mRNA and Cx32 protein Cx32 mRNA were weakly positive or negative in the cicatricial carcinoma epithelium. Compared with normal skin group and skin scar group, there were significant differences in positive area and average optical density between scar carcinoma group and normal skin group. The expression of Cx26 mRNA in pathological scar epithelium was significantly higher than that in normal skin group, and the positive expression of Cx26 mRNA in hypertrophic scar epithelium was significantly higher than that in normal skin group. The expression of Cx32mRNA (positive area and mean optical density) was significantly different (P 0.01). The expression of Cx32mRNA was positive in the pathological scar epithelium, compared with that in the scar carcinoma group. Compared with normal skin group, there was no significant difference in point mutation of Cx26 gene: after PCR reaction, 7 cases of target bands were amplified in 28 patients, after PCR reaction, there was no significant difference in the expression of Cx26 gene between the positive surface and the average optical density of the product (P 0.01), and there was no significant difference between the positive surface and the average optical density of the product. In 6 cases of scar carcinoma, the Cx26 gene of 4 cases was detected to have benign mutation by sequencing analysis, which was the point mutation analysis of two different types of polymorphic base mutation. Cx32 gene. After PCR reaction, 28 samples were detected. The target bands were amplified in 6 cases, of which 4 cases were scar carcinoma and 2 cases were scar carcinoma. No pathogenicity and non-pathogenic mutation sites were found by sequencing analysis. Conclusion the negative or weak positive expression of Cx26, mRNA and Cx32 and their Cx32mRNA in scar carcinoma may be associated with the loss of tumor growth function, which may lead to the occurrence of scar carcinoma. The strong positive and high expression of Cx26 and mRNA in scar epithelium. The decrease of Cx32mRNA expression in scar epithelium may be an early event of scar carcinogenesis. It is worthy to pay attention to the pathogenicity of Cx26Cx32 gene in the development of cicatricial carcinoma, and its significance needs to be further explored.
【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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