本文選題：皮膚鱗狀細胞癌 + DNA甲基化　； 參考：《新疆醫(yī)科大學》2014年博士論文

【摘要】：目的：檢測Wnt信號通路中SFRPs家族SFRP1，SFRP2，SFRP4，SFRP5基因啟動子在皮膚鱗狀細胞癌、癌旁組織和正常皮膚組織中甲基化率，探討SFRPs家族在皮膚鱗癌中甲基化狀態(tài)及其意義。同時檢測三組研究對象中Axin1、CyclinD1mRNA的表達，以及Wnt1和SFRP1蛋白在皮膚鱗癌和正常組織表達，探討Wnt信號通路中Axin1、Cyclin D1mRNA和Wnt1、SFRP1蛋白在皮膚鱗癌的作用，闡明SFRPs家族甲基化狀態(tài)與四個因子之間的關系及其在皮膚鱗癌中的發(fā)病機制。方法：（1）運用MassARRAY質(zhì)譜儀EpiTYPER甲基化方法檢測40例皮膚鱗癌，癌旁組織和正常皮膚組織SFRP1，SFRP2，SFRP4，SFRP5基因啟動子甲基化狀態(tài)；（2）運用實時熒光定量RT-PCR（Taqman probe）法檢測Axin1和Cyclin D1mRNA在皮膚鱗癌，癌旁組織和正常皮膚組織中的表達；（3）運用免疫組化SP法和Wertenblot方法檢測皮膚鱗癌與正常皮膚組織中Wnt1和SFRP1蛋白的表達；（4）運用SPSS17.0軟件對皮膚鱗癌，癌旁組織和正常皮膚組織中Wnt1、SFRP1蛋白、Axin1和Cyclin D1mRNA的表達及其與SFRP1，SFRP2，SFRP4，SFRP5甲基化狀態(tài)相關性進行分析，并分析其與皮膚鱗癌臨床病理學特征的關系，繪制SFRPs家族ROC曲線。結(jié)果：（1）皮膚鱗癌組織、癌旁組織、正常組織中SFRP1，SFRP2，SFRP4，SFRP5基因所研究CpG單位總數(shù)為7800個，其中6801個CpG單位可被分析（87.19%），均檢測出SFRPs家族甲基化狀態(tài)在三組研究對象中具有統(tǒng)計學差異的CpG位點以及在不同皮膚鱗癌病理級別具有統(tǒng)計學差異的CpG位點；（2）皮膚鱗癌組織中Axin1mRNA表達低于癌旁組織和正常皮膚組織，Cyclin D1mRNA表達高于癌旁組織和正常組織，Cyclin D1mRNA在不同病理分級表達差異具有統(tǒng)計學意義，Axin1mRNA在皮膚鱗癌不同病理分級表達無統(tǒng)計學意義。Axin1mRNA在皮膚鱗癌組織中呈低表達，Cyclin D1mRNA在皮膚鱗癌組織中呈高表達，兩者表達在皮膚鱗癌組織和癌旁組織中無相關性；（3）皮膚鱗癌組織中SFRPs家族檢測到31個CpG位點單位甲基化率與Axin1mRNA表達呈負相關；24個CpG位點單位甲基化率與Cyclin D1mRNA表達呈正相關；（4）免疫組化結(jié)果表明，Wnt1染色主要位于細胞質(zhì)呈棕黃色顆粒，Wnt1在皮膚鱗癌中表達陽性率明顯高于正常組織；SFRP1蛋白以細胞質(zhì)內(nèi)出現(xiàn)棕黃色顆粒染色為陽性（核中亦有少量表達），SFRP1在皮膚鱗癌組織表達陽性率低于正常皮膚組織。Wnt1蛋白表達與皮膚鱗癌組織病理有關，，CSCCⅢ級＞Ⅱ級＞Ⅰ級；SFRP1蛋白在不同病理分級的表達無差異。檢測SFRP1有7CpG位點基因高甲基化率與SFRP1蛋白表達呈負相關，與Wnt1蛋白表達呈正相關的8個CpG位點。結(jié)論：（1）檢測出SFRP1，SFRP2，SFRP4，SFRP5基因甲基化率在三組研究對象差異具有統(tǒng)計學意義的CpG位點，同時也檢測出在皮膚鱗癌不同病理分級中SFRP1，SFRP2，SFRP4，SFRP5基因甲基化率有統(tǒng)計學差異的CpG位點，提示SFRPs家族甲基化狀態(tài)可能在皮膚鱗癌發(fā)生與演進中起重要的作用，這些CpG位點為皮膚鱗癌表觀遺傳學研究提供理論依據(jù)；（2）Axin1mRNA表達下調(diào)和Cyclin D1mRNA表達增高可能與SFRPs家族在皮膚鱗癌組織中呈高甲基化狀態(tài)有關，Cyclin D1可作為惡性轉(zhuǎn)化的指標之一對臨床早期診斷，基因治療及預后判斷具有指導意義，兩者之間的關系仍需進一步探討；（3）Wnt1與SFRP1在皮膚鱗癌和正常組織中表達差異，與鱗癌組織的病理分級有關，提示W(wǎng)nt1與SFRP1可能參與了皮膚鱗癌的發(fā)生發(fā)展。SFRP1基因啟動子高甲基化狀態(tài)引起其基因失活導致染色質(zhì)結(jié)構(gòu)異常，從而SFRP1蛋白表達減少或缺失；（4）SFRP1中CpG1_1位點高甲基化率對皮膚鱗癌的識別率更高，為皮膚鱗癌早期診斷提供一定的理論基礎。
[Abstract]:Objective: to detect the methylation rate of SFRPs family SFRP1, SFRP2, SFRP4, SFRP5 gene promoter in skin squamous cell carcinoma, para cancerous tissue and normal skin tissue in Wnt signaling pathway, and to explore the methylation status and significance of SFRPs family in skin squamous cell carcinoma, and to detect the expression of Axin1, CyclinD1mRNA, Wnt1 and SFRP1 in the three groups of subjects. Expression of protein in skin squamous cell carcinoma and normal tissue, the role of Axin1, Cyclin D1mRNA and Wnt1, SFRP1 protein in skin squamous cell carcinoma in Wnt signaling pathway, and to elucidate the relationship between the methylation status of the SFRPs family and the four factors and the pathogenesis in the skin squamous cell carcinoma. Methods: (1) the detection of 40 by the EpiTYPER methylation of MassARRAY mass spectrometry (MassARRAY mass spectrometry). Cases of skin squamous cell carcinoma, paracancerous tissue and normal skin tissue SFRP1, SFRP2, SFRP4, SFRP5 promoter methylation status; (2) the expression of Axin1 and Cyclin D1mRNA in skin squamous cell carcinoma, para cancerous tissue and normal skin tissue was detected by real-time fluorescent quantitative RT-PCR (Taqman probe) method; (3) immunohistochemical SP method and Wertenblot method were used to detect the expression of Axin1 and Cyclin D1mRNA. Expression of Wnt1 and SFRP1 protein in skin squamous cell carcinoma and normal skin tissue; (4) the expression of Wnt1, SFRP1 protein, Axin1 and Cyclin D1mRNA in skin squamous cell carcinoma, paracancerous tissue and normal skin tissue by SPSS17.0 software and its correlation with SFRP1, SFRP2, SFRP4, SFRP5 methylation status, and the clinical pathology of skin squamous cell carcinoma were analyzed. The SFRPs family ROC curves were plotted. Results: (1) the total number of CpG units in the skin squamous cell carcinoma tissue, the para cancer tissue and the normal tissues SFRP1, SFRP2, SFRP4, SFRP5 genes was 7800, of which 6801 CpG units could be analyzed (87.19%), and all the SFRPs family methylation status had statistical difference in the three groups of research objects. The site and the CpG loci with statistical difference in the pathological grade of different skin squamous cell carcinoma; (2) the expression of Axin1mRNA in skin squamous cell carcinoma is lower than that of para cancer tissue and normal skin tissue, and the expression of Cyclin D1mRNA is higher than that of para cancer tissue and normal tissue. The difference of Cyclin D1mRNA in different pathological grades is statistically significant, Axin1mRNA is in the skin. The expression of.Axin1mRNA in squamous cell carcinoma was low expression in squamous cell carcinoma, and Cyclin D1mRNA was highly expressed in skin squamous cell carcinoma tissue. There was no correlation between the expression of Cyclin D1mRNA in skin squamous cell carcinoma tissues and adjacent tissues. (3) the SFRPs family in skin squamous cell carcinoma detected the methylation rate and Axin1mRNA in the unit of CpG loci. The expression was negatively correlated; the unit methylation rate of 24 CpG loci was positively correlated with the expression of Cyclin D1mRNA. (4) the results of immunohistochemical staining showed that Wnt1 staining was mainly located in the cytoplasm of brown yellow granules, and the positive rate of Wnt1 in skin squamous cell carcinoma was significantly higher than that of normal tissue; SFRP1 protein was stained by brown yellow granules in cytoplasm as positive (nucleus). The positive rate of SFRP1 in skin squamous cell carcinoma was lower than that of normal skin tissue. The expression of.Wnt1 protein was related to the histopathology of skin squamous cell carcinoma, CSCC grade III > II > grade I, and there was no difference in the expression of SFRP1 protein in different pathological grades. There was a negative correlation between the high methylation rate of 7CpG site and the expression of SFRP1 protein in SFRP1. 8 CpG loci positively correlated with the expression of Wnt1 protein. Conclusion: (1) the methylation rate of SFRP1, SFRP2, SFRP4, SFRP5 gene was detected in the three groups with statistically significant CpG loci, and the allele of SFRP1, SFRP2, SFRP4 and SFRP5 genes in different pathological grades of skin squamous cell carcinoma was also detected. It is suggested that the methylation status of the SFRPs family may play an important role in the development and evolution of skin squamous cell carcinoma. These CpG loci provide a theoretical basis for the epigenetic study of skin squamous cell carcinoma. (2) the downregulation of Axin1mRNA expression and the increase of Cyclin D1mRNA expression may be related to the hypermethylation status of the SFRPs family in the skin squamous cell carcinoma, Cyclin D1 As one of the indicators of malignant transformation, it is of guiding significance for early clinical diagnosis, gene therapy and prognosis, and the relationship between them still needs further discussion. (3) the difference between Wnt1 and SFRP1 in skin squamous cell carcinoma and normal tissues is related to the pathological grading of squamous cell carcinoma, suggesting that Wnt1 and SFRP1 may be involved in the occurrence of squamous cell carcinoma of the skin. The development of the hypermethylation of.SFRP1 gene promoter causes its gene inactivation to lead to abnormal chromatin structure, thus reducing or missing the expression of SFRP1 protein. (4) the high methylation rate of CpG1_1 loci in SFRP1 provides a higher recognition rate for skin squamous cell carcinoma and provides a theoretical basis for the early diagnosis of skin squamous cell carcinoma.
【學位授予單位】：新疆醫(yī)科大學
【學位級別】：博士
【學位授予年份】：2014
【分類號】：R739.5

【相似文獻】

相關期刊論文 前10條

1 劉云貞;楊頂權(quán);白彥萍;;手術切除頭部皮膚鱗癌1例[J];實用皮膚病學雜志;2010年03期

2 許俏;羅俊;潘年松;;中藥抗皮膚鱗癌的體外實驗機制探討[J];中國醫(yī)藥指南;2012年29期

3 戶曉紅;左足巂皮膚鱗癌伴左鎖骨上淋巴結(jié)轉(zhuǎn)移—例[J];實用癌癥雜志;1995年04期

4 李素卿,成志明,張仁亞;多發(fā)性皮膚鱗癌1例[J];中國皮膚性病學雜志;1997年01期

5 張淑娟;劉寧;代曉明;李逸松;;皮膚鱗癌動物模型構(gòu)建[J];昆明醫(yī)學院學報;2009年S2期

6 劉寧;張淑娟;代曉明;李逸松;;紫外線引發(fā)皮膚鱗癌的機制研究[J];昆明醫(yī)學院學報;2009年S2期

7 朱楨;曹海鵬;齊鳳琴;張素華;;腫瘤抑癌基因PTEN蛋白在皮膚鱗癌中的表達及其意義[J];中國誤診學雜志;2011年15期

8 匡玉珍;肖新華;;結(jié)締組織生長因子的表達與皮膚鱗癌臨床分期和病理分級的關系[J];中南醫(yī)學科學雜志;2011年04期

9 吳夢濤;袁瑞敏;趙梅;;皮膚鱗癌轉(zhuǎn)移并發(fā)下肢靜脈血栓1例[J];中華全科醫(yī)學;2012年05期

10 衛(wèi)江華;關哲;;60例皮膚鱗癌綜合治療分析[J];山西職工醫(yī)學院學報;2012年03期

相關會議論文 前10條

1 楊欣;馬勇光;張潔;尤維濤;侯俊杰;余若輝;郭亮;王俠;李東;李健寧;;游離股前外側(cè)皮瓣修復巨大皮膚鱗癌切除后缺損[A];中華醫(yī)學會整形外科學分會第十一次全國會議、中國人民解放軍整形外科學專業(yè)委員會學術交流會、中國中西醫(yī)結(jié)合學會醫(yī)學美容專業(yè)委員會全國會議論文集[C];2011年

2 楊欣;馬勇光;張潔;尤維濤;侯俊杰;余若輝;郭亮;王俠;李東;李健寧;;游離股前外側(cè)皮瓣修復巨大皮膚鱗癌切除后缺損[A];中華醫(yī)學會整形外科學分會第十一次全國會議、中國人民解放軍整形外科學專業(yè)委員會學術交流會、中國中西醫(yī)結(jié)合學會醫(yī)學美容專業(yè)委員會全國會議論文集[C];2011年

3 崔榮;馮捷;閆小寧;李文彬;張彩晴;胡剛;史丙俊;魏玉萍;曹偉;;腫瘤-成纖維細胞相互作用誘導肌成纖維細胞分化及對皮膚鱗癌生長侵襲作用的研究[A];中華醫(yī)學會第14次全國皮膚性病學術年會論文匯編[C];2008年

4 趙娟;顏蘭香;;抑癌基因FHIT在皮膚鱗癌中的表達及其甲基化狀態(tài)[A];中華醫(yī)學會第14次全國皮膚性病學術年會論文匯編[C];2008年

5 王珍;何春滌;陳洪鐸;王雅坤;肖汀;;維甲酸、γ-干擾素誘導皮膚鱗癌細胞發(fā)生細胞凋亡的機制研究[A];中華醫(yī)學會第十二次全國皮膚性病學術會議論文集[C];2006年

6 賈近博;林盡染;陳國梁;徐金華;陳明華;;羥基脲相關多發(fā)疣狀皮膚角化并發(fā)皮膚鱗癌一例[A];中華醫(yī)學會第16次全國皮膚性病學術年會摘要集[C];2010年

7 鄭焱;;皮膚鱗癌中DBPA的升高及其與表皮分化的關系[A];2010全國中西醫(yī)結(jié)合皮膚性病學術會議論文匯編[C];2010年

8 王建力;張捷;;皮膚鱗癌中E-cad表達與微血管密度相關性的研究[A];中華醫(yī)學會第十二次全國皮膚性病學術會議論文集[C];2006年

9 段志武;史景祿;王成;;皮膚鱗癌中的Bcl-2表達[A];2001年中國中西醫(yī)結(jié)合皮膚性病學術會議論文匯編[C];2001年

10 李泓馨;;皮膚鱗癌A431腫瘤干細胞的檢測以及初步鑒定[A];中華醫(yī)學會第十八次全國皮膚性病學術年會論文匯編[C];2012年

相關博士學位論文 前1條

1 梁俊琴;皮膚鱗癌組織中SFRPs家族甲基化狀態(tài)及其作用機制的研究[D];新疆醫(yī)科大學;2014年

相關碩士學位論文 前10條

1 趙思成;熊果酸對人皮膚鱗癌的抑制作用及機制研究[D];南京醫(yī)科大學;2010年

2 張淑娟;紫外線誘導裸鼠皮膚鱗癌動物模型的構(gòu)建[D];昆明醫(yī)學院;2011年

3 宣敏;基質(zhì)金屬蛋白酶-12及其組織抑制物-1在皮膚鱗癌中的表達研究[D];南方醫(yī)科大學;2011年

4 楊陽;GSK3β調(diào)節(jié)細胞自噬在皮膚鱗癌發(fā)病中的作用及分子機制[D];第四軍醫(yī)大學;2012年

5 余秀琴;調(diào)控NPM對人皮膚鱗癌細胞增殖活性的影響研究[D];蘇州大學;2013年

6 李桂寅;人類皰疹病毒8型ORF26和ORF75基因亞型在鱗狀細胞癌中的分布[D];石河子大學;2011年

7 石磊;5-氨基酮戊酸聚乳酸羥基乙酸納米粒的制備及其光動力療法抑制人皮膚鱗癌A-431細胞體外增殖研究[D];安徽醫(yī)科大學;2012年

8 高昆;腺病毒介導的PKCδ基因?qū)ζつw鱗癌細胞抑制作用的實驗研究[D];大連醫(yī)科大學;2013年

9 于文心;HIF-1α、MMP-2在皮膚鱗癌中的表達[D];青島大學;2012年

10 張云鳳;一氧化氮在ALA光動力誘導人皮膚鱗癌A431細胞凋亡中的作用研究[D];揚州大學;2013年

本文編號：2039550