本文關(guān)鍵詞：皮膚鱗狀細(xì)胞癌與基底細(xì)胞癌差異蛋白質(zhì)組學(xué)研究　出處：《鄭州大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

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【摘要】：背景與目的 鱗狀細(xì)胞癌和基底細(xì)胞癌均為常見(jiàn)的皮膚惡性腫瘤，它們都起源于表皮或皮膚附屬器，具有相近的好發(fā)部位和發(fā)病機(jī)制，并且兩種疾病在發(fā)病部位的局部也有相似的臨床表現(xiàn)。但在腫瘤發(fā)生轉(zhuǎn)移的情況上，兩種疾病有著較大差異。 蛋白質(zhì)組學(xué)（proteomics）是指將細(xì)胞或組織中的蛋白質(zhì)組作為整體進(jìn)行研究，分析蛋白質(zhì)組中各蛋白的成分組成以及相互之間的的影響。蛋白質(zhì)組學(xué)主要是對(duì)蛋白質(zhì)組的蛋白質(zhì)構(gòu)成、差異蛋白質(zhì)組學(xué)研究以及在生物活動(dòng)中各蛋白質(zhì)之間發(fā)揮的作用這三個(gè)方面進(jìn)行主要的研究。 這次實(shí)驗(yàn)應(yīng)用蛋白質(zhì)組學(xué)的相關(guān)技術(shù)對(duì)人皮膚鱗狀細(xì)胞癌細(xì)胞和基底細(xì)胞癌細(xì)胞中所提取的蛋白質(zhì)進(jìn)行分析，找出其中存在差異的蛋白質(zhì)，并對(duì)部分差異蛋白進(jìn)行功能驗(yàn)證。這將對(duì)皮膚鱗狀細(xì)胞癌、基底細(xì)胞癌的早期疾病診斷、早期發(fā)生及轉(zhuǎn)移等腫瘤標(biāo)志物的鑒定、腫瘤治療特異性位點(diǎn)的篩選，以及新的臨床治療方案的制定均會(huì)有重要的意義。 方法 1細(xì)胞的培養(yǎng)與樣品蛋白的提取本實(shí)驗(yàn)選擇人皮膚鱗狀細(xì)胞癌A-431細(xì)胞系和人基底細(xì)胞癌TE-354.T做為腫瘤細(xì)胞模型進(jìn)行常規(guī)細(xì)胞培養(yǎng)。分別取對(duì)數(shù)生長(zhǎng)期的兩種細(xì)胞，待細(xì)胞貼壁生長(zhǎng)至培養(yǎng)瓶培養(yǎng)面面積的90%時(shí)，進(jìn)行樣本蛋白提取。并應(yīng)用Bradford法對(duì)樣品蛋白濃度進(jìn)行測(cè)定。 2樣品蛋白的雙向凝膠電泳通過(guò)IPG干膠條重水化、等電聚焦電泳、平衡IPG膠條、垂直向SDS-PAGE電泳、顏色等步驟分別對(duì)兩種樣品蛋白質(zhì)進(jìn)行蛋白分離，每個(gè)樣品分別制3塊雙向電泳膠板，并應(yīng)用圖像掃描儀進(jìn)行掃描，獲得兩種蛋白樣品的雙向凝膠電泳圖譜。用圖像分析軟件對(duì)雙向凝膠電泳圖譜進(jìn)行分析，找出差異蛋白質(zhì)點(diǎn)。 結(jié)果 1兩種細(xì)胞系提取蛋白的雙向電泳蛋白圖譜大眼管基本一致，各個(gè)蛋白質(zhì)著色點(diǎn)基本都聚集在等電點(diǎn)（Ip）為pH4~7的范圍內(nèi)；分子量（Ms）大多在15kDa~150kDa的范圍內(nèi)。 2通過(guò)對(duì)在皮膚鱗狀細(xì)胞癌細(xì)胞系和皮膚基底細(xì)胞癌細(xì)胞系中提取樣品蛋白的雙向凝膠電泳膠板的分析，其中有16.8%的蛋白質(zhì)點(diǎn)存在差異，其中13個(gè)蛋白在鱗狀細(xì)胞癌細(xì)胞中表達(dá)上調(diào)；和2個(gè)在鱗狀癌細(xì)胞細(xì)胞表達(dá)下調(diào)的蛋白質(zhì)。4個(gè)蛋白點(diǎn)在基底細(xì)胞癌組中未檢測(cè)出。 結(jié)論 1皮膚鱗狀細(xì)胞癌細(xì)胞系和皮膚基底細(xì)胞癌細(xì)胞系中提取樣品蛋白的雙向凝膠電泳圖譜中存在部分差異。皮膚鱗狀細(xì)胞癌所含蛋白質(zhì)組分較基底細(xì)胞癌的復(fù)雜。 2分子量和等電點(diǎn)分別為（22.32，3.95）、（27.48，4.3）、（58.96，7.45）、（64.8，5）、（121.8，，5.84）、（135.13，6.44）等蛋白質(zhì)的表達(dá)差異可能是造成鱗狀細(xì)胞癌和基底細(xì)胞癌在腫瘤轉(zhuǎn)移方面有較大差異的主要影響因素。
[Abstract]:Background and Purpose Squamous cell carcinoma and basal cell carcinoma are common malignant tumor of the skin , all of them originate from the epidermis or the skin accessory , have similar good hair parts and pathogenesis , and the two diseases have similar clinical manifestations at the site of the disease . But in the case of tumor metastasis , the two diseases have a large difference . Proteomics refers to the study of the composition of each protein in the protein group and its influence on each other . Proteomics is mainly used to study the structure of protein in the proteome , the differential proteomic study and the role of the proteins in the biological activity . In this experiment , the proteins extracted from human skin squamous cell carcinoma cells and basal cell carcinoma cells were analyzed by using the technique of Proteomics . It was found that there were different proteins and functionally validated some of the difference proteins . This would be of great significance to the identification of tumor markers such as squamous cell carcinoma of skin , early diagnosis of basal cell carcinoma , early occurrence and metastasis , screening of tumor treatment specific sites , and the development of new clinical treatment programmes . method Human skin squamous cell carcinoma A - 431 cell line and human basal cell carcinoma TE - 354.T were used as tumor cell model to carry out normal cell culture . The two - way gel electrophoresis of two kinds of sample proteins was carried out by two - way gel electrophoresis of two sample proteins , such as rehydration , isoelectric focusing , SDS - PAGE electrophoresis , color and so on . The two - way gel electrophoresis map of two kinds of protein samples was obtained by scanning with image scanner . The two - way gel electrophoresis map was analyzed by image analysis software to find out the difference protein spots . Results The isoelectric point ( Ip ) of each protein staining point is within the range of pH 4 - 7 , and the molecular weight ( Ms ) is mostly in the range of 15 kDa to 150 kDa . 2 . Analysis of two - way gel electrophoresis gel electrophoresis for the extraction of sample proteins in cancer cell lines and basal cell carcinoma cells of the skin . There were 16.8 % of the protein spots , 13 of which were up - regulated in squamous cell carcinoma cells , and 2 proteins down - regulated in squamous cell carcinoma cells . Four protein spots were not detected in basal cell carcinoma . Conclusion There are some differences in the two - way gel electrophoresis of the sample protein extracted from the cancer cell line of skin squamous cell carcinoma and basal cell line of skin . The protein component of squamous cell carcinoma of skin is more complex than basal cell carcinoma . The molecular weight and isoelectric point were ( 22.32 , 3.95 ) , ( 27.48 , 4.3 ) , ( 58.96 , 7.45 ) , ( 64.8 , 5 ) , ( 121.8 , 5.84 ) , ( 135.13 , 6.44 ) and so on .

【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R739.5

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