本文選題：Cx26 + Cx32　； 參考：《遵義醫(yī)學(xué)院》2011年碩士論文

【摘要】：目的：探討Cx26、Cx32蛋白和分子水平的表達、基因突變在病理性皮膚瘢痕及瘢痕癌中的意義。方法：以病理性皮膚瘢痕、皮膚瘢痕癌組織為研究對象,以正常皮膚組織為對照。采用免疫組織化學(xué)(S-P)法分別檢測Cx26、Cx32蛋白的表達,采用核酸分子原位雜交法檢測Cx26mRNA、Cx32mRNA的表達,結(jié)合圖像分析,分別觀測三組被檢組織中所檢各項指標的表達(陽性面積與平均光密度),所有數(shù)據(jù)輸入計算機后運用SPSS16.0軟件包進行統(tǒng)計學(xué)分析；從14例皮膚瘢痕和14例瘢痕癌石蠟包埋組織中提取DNA,運用PCR擴增及擴增目的片段直接測序的方法,進行突變篩查。結(jié)果：(1)Cx26蛋白、Cx26mRNA和Cx32蛋白、Cx32mRNA在瘢痕癌上皮中呈弱陽性或陰性表達。瘢痕癌組分別與正常皮膚組、皮膚瘢痕組比較,表達(陽性面積與平均光密度)差異均有統(tǒng)計學(xué)意義(P0.01)；(2)Cx26蛋白、Cx26mRNA在病理性瘢痕上皮中呈強陽性表達,與正常皮膚組比較,其表達(陽性面積與平均光密度)差異有統(tǒng)計學(xué)意義(P0.01); (3) Cx32mRNA在病理性瘢痕上皮的表達呈陽性,與瘢痕癌組比較,其表達(陽性面與積平均光密度)差異有統(tǒng)計學(xué)意義(P0.01),與正常皮膚組比較,差異無統(tǒng)計學(xué)意義(P0.05)。(4)Cx26基因點突變分析：經(jīng)PCR反應(yīng)后,在28例患者樣本中,擴增出7例目的條帶,其中6例瘢痕1例瘢痕癌,經(jīng)測序分析檢測到4例瘢痕的Cx26基因有良性突變,為兩種不同類型的多態(tài)性堿基變異。(5)Cx32基因點突變分析：經(jīng)PCR反應(yīng)后,在28例樣本中,擴增出6例目的條帶,其中4例瘢痕2例瘢痕癌,經(jīng)測序分析均未發(fā)現(xiàn)致病性與非致病性突變位點。結(jié)論：(1)瘢痕癌中Cx26及其mRNA和Cx32及其Cx32mRNA的陰性或弱陽性、低表達,可能與其抑制腫瘤生長功能的喪失,導(dǎo)致瘢痕癌的發(fā)生有相關(guān)性；(2)瘢痕上皮中Cx26及其mRNA的強陽性、高表達,可能與促進瘢痕上皮增生有相關(guān)性,同時也與病理性瘢痕易形成潰瘍、潰瘍經(jīng)久不愈有相關(guān)性；(3)瘢痕上皮中Cx32mRNA表達水平的降低,有可能是瘢痕癌變的早期事件,值得重視；(4) Cx26、Cx32基因在瘢痕癌的發(fā)生發(fā)展過程中尚未發(fā)現(xiàn)有致病性的基因突變位點,其意義有待進一步探討。
[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.
【學(xué)位授予單位】：遵義醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R739.5

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