本文選題：口腔白斑惡變 + 口腔鱗癌　； 參考：《大連醫(yī)科大學(xué)》2014年碩士論文

【摘要】：口腔黏膜白斑（oral leukoplakia，OLK）是口腔黏膜上的白色斑塊或白色斑片，不能以臨床和組織病理學(xué)的方法診斷為其他任何疾病者。OLK惡變率高，可惡變成嚴(yán)重危害人類生命健康的口腔黏膜鱗狀細胞癌（oral squamous cellcarcinoma，OSCC)，其中非均質(zhì)型白斑惡變率約為13%-17.5%，甚至在某些地區(qū)OLK惡變率可高達50%-70%。口腔黏膜鱗狀細胞癌是世界十大腫瘤之一，在全身惡性腫瘤中排第六位。有研究結(jié)果顯示，口腔鱗狀細胞癌中有17%-35%是由口腔白斑惡變而來，因此研究口腔白斑惡變分子機制，篩選惡變相關(guān)蛋白，尋找惡變分子標(biāo)志物，對口腔白斑惡變的早期發(fā)現(xiàn)、早期診斷及治療，有效改善預(yù)后具有重要的科學(xué)意義和臨床應(yīng)用前景。 目的：本項研究通過蛋白質(zhì)組學(xué)檢測技術(shù)篩檢口腔黏膜白斑惡變相關(guān)蛋白，并分析OLK惡變前后差異蛋白的相關(guān)信息，為深入研究口腔黏膜白斑惡變分子機制，探尋口腔白斑惡變分子標(biāo)志物及關(guān)鍵治療靶點提供重要的研究基礎(chǔ)。 方法：本實驗采用我們自行建立的口腔黏膜白斑惡變細胞模型，即人口腔黏膜白斑細胞系DOK及其誘導(dǎo)惡變細胞系口腔鱗狀上皮癌細胞OSCC-B+D為實驗對象，運用相對定量蛋白質(zhì)組學(xué)方法高通量篩檢DOK及OSCC-B+D細胞間蛋白質(zhì)組的表達差異。首先采用穩(wěn)定同位素二甲基標(biāo)記技術(shù)，將CH2O和CD2O分別標(biāo)記進入人口腔白斑細胞DOK-G及其惡變細胞OSCC-B+D的蛋白酶解肽段，然后將兩種細胞樣品混合通過高效液相色譜進行分離，最后樣品進入具有高靈敏性、可微量上樣的Orbitrap Elite質(zhì)譜儀進行多肽檢測，并通過MaxQuant軟件（v1.1.2.5），在IPIHuman.（v3.87）數(shù)據(jù)庫中對質(zhì)譜檢測結(jié)果進行蛋白質(zhì)鑒定。 結(jié)果：口腔黏膜白斑細胞DOK及其惡變細胞OSCC-B+D樣品經(jīng)過HPLC-MS/MS檢測分析后，共鑒定出1459個蛋白，取三次重復(fù)實驗的共同鑒定結(jié)果，篩檢出931個蛋白。對獲得的篩檢結(jié)果進行進一步過濾，控制相對標(biāo)準(zhǔn)偏差RSD＜35%，最終共獲得621個蛋白。分析和比較DOK及其惡變細胞OSCC-B+D蛋白質(zhì)組的表達變化，篩選出197個惡變前后的差異蛋白，其中125個蛋白在惡變后呈顯著性上調(diào)表達（平均氘氫比值2），72個蛋白在惡變后呈顯著性下調(diào)表達（平均氘氫比值0.5）。 我們采用uniprotKB數(shù)據(jù)庫將197個差異蛋白按照其細胞內(nèi)定位，蛋白分子功能及其參與的生物學(xué)過程進行分類分析，獲得差異蛋白相關(guān)信息。依據(jù)細胞內(nèi)定位信息，提示差異蛋白中定位于細胞質(zhì)（cytoplasm）和細胞核（nucleus）的蛋白最多，均占22%，其他依次遞減定位于線粒體（mitochondrion，12%），細胞膜（plasma membrane，11%），內(nèi)質(zhì)網(wǎng)（endoplasmic reticulum，8%），integralto membrane（7%）；依據(jù)蛋白分子功能信息，提示DNA結(jié)合（DNA binding）蛋白最多，約占9%,其次是RNA結(jié)合（RNA binding）蛋白，約占6%,其他依次遞減為載體活性（transporter activity）蛋白（5%）,GTP酶調(diào)節(jié)活性（GTPase activity，3%）蛋白，轉(zhuǎn)錄監(jiān)管活性（transcription regulator activity，2%）蛋白等；依據(jù)參與的生物學(xué)過程信息，提示參與核糖核酸/脫氧核糖核酸/核苷酸/核小體的代謝調(diào)節(jié)（regulation of RNA/DNA/nucleotide/nucleosome metabolic process）蛋白最多約占23%，其次為蛋白代謝（protein metabolism）約占12%，其他依次遞減為轉(zhuǎn)運（transport，11%）蛋白，細胞生長（cell growth，9%），細胞凋亡（apoptosis，7%），信號轉(zhuǎn)導(dǎo)（signal transduction，2%）以及免疫反應(yīng)（immuneresponse，，2%）相關(guān)蛋白。 結(jié)論：本項研究通過高通量篩檢口腔黏膜白斑細胞惡變前后的差異蛋白質(zhì)組，發(fā)現(xiàn)197個蛋白質(zhì)呈差異表達。根據(jù)差異蛋白相關(guān)信息，初步推測OLK惡變發(fā)生的可能機制：參與OLK惡變發(fā)生的蛋白大多定位于細胞核與細胞質(zhì)中，DNA/RNA結(jié)合蛋白及參與核糖核酸/脫氧核糖核酸/核苷酸/核小體代謝調(diào)節(jié)的蛋白質(zhì)在OLK惡變進程中發(fā)揮重要作用，DNA/RNA功能及代謝異常可能是OLK惡變的重要機制。本項研究為進一步深入細致地探索OLK惡變分子機制，篩選口腔黏膜白斑惡變分子標(biāo)志物和關(guān)鍵治療靶點提供有力的研究基礎(chǔ)和科學(xué)依據(jù)。 目的：通過對大學(xué)生進行口腔健康檢查，了解這一人群的齲病患病情況，為制訂口腔保健治療方案提供科學(xué)依據(jù)，同時充實大學(xué)生齲病流行病學(xué)資料。 方法：參照WHO口腔健康調(diào)查基本方法以及第三次全國口腔健康流行病學(xué)調(diào)查方案制定口腔檢查表格，對大連理工大學(xué)570名大學(xué)生進行齲齒患病情況研究，調(diào)查內(nèi)容包括齲均、患齲率等,以視診結(jié)合探診的方式檢查后將結(jié)果記錄在表格內(nèi)。 結(jié)果：570名大學(xué)生患齲率為58.07%，齲失補牙數(shù)共1013顆牙（齲牙數(shù)766顆，因齲失牙數(shù)10顆，因齲補牙數(shù)237顆），齲均為1.78±2.335，齲齒充填率為23.40%。男生和女生在患齲率分別為54.3%和73.5%，兩者差異具有統(tǒng)計學(xué)意義（P＜0.05）；男生和女生的齲均分別為1.54±2.130和2.73±2.845，兩者差異具有統(tǒng)計學(xué)意義（P＜0.05）。城市生源和農(nóng)村生源學(xué)生的患齲率分別為62.1%和53.2%，兩者差異具有統(tǒng)計學(xué)意義（P＜0.05）；城市生源和農(nóng)村生源的齲均分別為1.95±2.322和1.56±2.389，兩者差異無統(tǒng)計學(xué)意義（P＞0.05）。不同年齡分組的患齲率分別為58.5%（18-20歲）、54.9%（21-23歲）、64.3%（＞24歲），三者差異無統(tǒng)計學(xué)意義（P＞0.05）；不同年齡分組的齲均分別為1.72±2.108（18-20歲）、1.63±2.325（21-23歲）、2.36±2.968（＞24歲），三者組間比較差異具有統(tǒng)計學(xué)意義（P＜0.05）。對不同年齡分組的齲均進行兩兩比較后顯示18-20歲年齡段與21-23歲年齡段的齲均無統(tǒng)計學(xué)差異，但二者與＞24歲年齡組均有統(tǒng)計學(xué)差異，隨著年齡增長，齲均也隨之增加。進一步對患齲牙位進行分析比較之后，發(fā)現(xiàn)磨牙患齲水平明顯高于其他牙齒，其中下頜磨牙患齲率高于上頜磨牙，二者具有統(tǒng)計學(xué)差異（P＜0.05）。 結(jié)論：通過本次齲病流行病學(xué)調(diào)查，結(jié)果表明大學(xué)生齲齒患病率較高，但齲齒充填率較低。本次調(diào)查顯示大學(xué)生的齲病流行特點為：齲齒患病率女性高于男性、城市高于鄉(xiāng)村；齲均水平女性高于男性、24歲以上年齡組高于18-23歲年齡組�；箭x牙位的流行病特點為：磨牙患齲水平高于其他牙齒，下頜磨牙患齲水平高于上頜磨牙。
[Abstract]:Oral leukoplakia (oral leukoplakia, OLK) is a white plaque or white plaque on the oral mucosa. It can not be diagnosed as any other disease by clinical and histopathological methods, and the rate of malignant transformation of.OLK is high, and the evil becomes the oral squamous cellcarcinoma (OSCC), which is serious harm to human life and health (oral squamous cellcarcinoma, OSCC). The malignant change rate of white leukoplakia is about 13%-17.5%. Even in some areas, the rate of OLK malignant change can be as high as 50%-70%. oral mucosa squamous cell carcinoma is one of the ten largest tumors in the world, and it is ranked sixth in the malignant tumor of the whole body. The results of research show that the 17%-35% is malignant in oral squamous cell carcinoma, and therefore, the malignant change of oral leukoplakia is studied. Submechanism, screening of malignant change related proteins and finding malignant molecular markers, early detection of oral leukoplakia malignancy, early diagnosis and treatment, and effective improvement of prognosis have important scientific significance and clinical application prospects.
Objective: This study screened oral leukoplakia malignancy related proteins by proteomic technology and analyzed the related information of differential proteins before and after OLK malignancy, and provided an important research basis for the in-depth study of the molecular mechanism of oral leukoplakia and the exploration of biomarkers and key therapeutic targets for oral leukoplakia.
Methods: in this experiment, the oral leukoplakic leukoplakia malignant cell model, the human oral leukoplakic leukoplakia cell line DOK and the oral squamous cell carcinoma cell line OSCC-B+D induced by the malignant cell line, were used as the experimental object. The relative quantitative proteomics method was used to detect the differential expression of the protein group between DOK and OSCC-B+D cells. Firstly, CH2O and CD2O were used to label the CH2O and CD2O respectively into the protease peptide segments of the human oral leukoplakia cell DOK-G and its malignant cell OSCC-B+D, and then the samples were mixed by high performance liquid chromatography, and the final samples were highly sensitive, and the Orbitrap Elite could be found in the sample. The peptide was detected by mass spectrometer and identified by MaxQuant software (v1.1.2.5) in the IPIHuman. (v3.87) database.
Results: after HPLC-MS/MS detection and analysis of oral leukoplakia leukoplakia DOK and its malignant cell OSCC-B+D samples, 1459 proteins were identified and 931 proteins were screened out from three repeated experiments. The results of screening were further filtered, and the standard deviation was RSD < 35%, and 621 proteins were obtained. By analyzing and comparing the changes in the expression of DOK and its malignant cell OSCC-B+D proteome, 197 differential proteins were screened, of which 125 proteins were significantly up-regulated after the malignant change (the average deuterium hydrogen ratio 2), and the 72 proteins decreased significantly after the malignant change (the ratio of deuterium hydrogen was 0.5).
We used the uniprotKB database to classify the 197 differentially proteins according to their intracellular localization, protein molecular function and their biological processes to obtain differential protein related information. According to the intracellular location information, the most proteins located in the cytoplasm (cytoplasm) and the nucleus (nucleus) in the differential proteins were the most, which accounted for 22%. Others descended in sequence in mitochondria (mitochondrion, 12%), cell membrane (plasma membrane, 11%), endoplasmic reticulum (endoplasmic reticulum, 8%), integralto membrane (7%). According to protein molecular function information, DNA binding (DNA binding) protein was most, accounting for about 9%, followed by RNA binding (RNA) protein, accounting for 6%, and the others descended successively in turn. Carrier activity (transporter activity) protein (5%), GTP enzyme regulatory activity (GTPase activity, 3%) protein, transcriptional regulatory activity (transcription regulator activity, 2%), etc., and in accordance with the biological process information involved in the metabolic regulation of ribonucleic acid / deoxynuclear nucleic acid / nucleosome (regulation of RNA/DNA/nucl) Eotide/nucleosome metabolic process) protein accounted for about 23%, followed by protein metabolism (protein metabolism) accounting for about 12%, others descended in turn to transport (transport, 11%) protein, cell growth (cell growth, 9%), apoptosis (apoptosis, 7%), signal transduction (signal transduction, 2%), and immune response (immuneresponse, 2%) related proteins.
Conclusion: in this study, 197 proteins were found to be differentially expressed in the differential proteome before and after the malignant transformation of leukoplakia cells in the oral mucosa. According to the information of differential protein, the possible mechanism of OLK malignant transformation was preliminarily deduced: most of the proteins involved in the OLK malignant transformation were located in the nucleus and cytoplasm, and DNA/RNA combined with eggs. The protein that Rhizoma Bletillae participates in RNA / deoxyribonucleic acid / nucleosome metabolic regulation plays an important role in the OLK malignant transformation process. The DNA/RNA function and metabolic abnormalities may be an important mechanism for the malignant transformation of OLK. This study is to further explore the OLK malignant molecular mechanism and screen the biomarkers of oral leukoplakia malignant molecules. The material and key therapeutic targets provide strong basis for research and scientific basis.
Objective: to understand the prevalence of dental caries in this population by oral health examination to college students, to provide a scientific basis for the formulation of oral health care treatment scheme and to enrich the epidemiological data of dental caries for college students.
Methods: according to the basic methods of the WHO oral health survey and the third national oral health epidemiological survey, 570 college students in Dalian University of Technology were studied for dental caries, including caries and caries. The results were recorded in the form after inspection. Inside.
Results: the caries rate of 570 college students was 58.07%, the number of dental caries was 1013 teeth (766 teeth, 10 teeth, 237 dental caries), and the caries were 1.78 + 2.335. The dental caries rate of 23.40%. boys and girls was 54.3% and 73.5%, respectively, and the difference was statistically significant (P < 0.05), and the caries of boys and girls were both. The differences were 1.54 + 2.130 and 2.73 + 2.845 respectively (P < 0.05). The rate of dental caries of urban and rural students were 62.1% and 53.2% respectively (P < 0.05), and the caries of urban and rural sources were 1.95 + 2.322 and 1.56 + 2.389, respectively (P > 0.05) the incidence of caries in different age groups was 58.5% (18-20 years old), 54.9% (21-23 years old) and 64.3% (> 24 years), and there was no significant difference in three (P > 0.05). The average caries in different age groups were 1.72 + 2.108 (18-20 years old), 1.63 + 2.325 (> 54.9% years old), and the differences were statistically significant (P <). There was no significant difference in caries between the age group of 18-20 years and the age group at the age of 21-23, but there was a statistical difference between the two and the 21-23 years old age group, but the caries increased with the age of 24. After the analysis and comparison of the dental caries, it was found that the level of dental caries was significantly higher than that of the 21-23. The incidence of dental caries in mandibular molars was higher than that in maxillary molars, and the difference between the two groups was statistically significant (P < 0.05).
Conclusion: through the epidemiological survey of caries, the prevalence rate of dental caries of college students is higher, but the rate of dental caries filling is low. This survey shows that the prevalence of dental caries in college students is higher than that of men, the city is higher than that of the countryside, the average caries level in women is higher than that of men, and the age group over 24 years old is higher than the age group of 18-23 years. The epidemiological characteristics of dental caries are: the level of molar caries is higher than that of other teeth, and the level of caries of mandibular molars is higher than that of maxillary molars.
【學(xué)位授予單位】：大連醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R739.85;R781.1

【參考文獻】

相關(guān)期刊論文 前10條

1 趙玉澤;王宏坤;鄭繪霞;;S100A4基因在腫瘤研究中的進展[J];山西醫(yī)藥雜志;2009年08期

2 汪一鳴,李良壽;口腔粘膜白斑病流行病學(xué)調(diào)查研究述評[J];實用口腔醫(yī)學(xué)雜志;1996年01期

3 李鳴宇,劉正;綠茶多酚和氟化鈉對變鏈菌形態(tài)學(xué)及酶學(xué)的影響[J];實用口腔醫(yī)學(xué)雜志;1999年03期

4 康舒平;;對87例口腔黏膜白斑病患者臨床特點的分析[J];求醫(yī)問藥(下半月);2013年01期

5 周愿;單亦初;張麗華;張玉奎;;基于穩(wěn)定同位素標(biāo)記的蛋白質(zhì)組學(xué)定量方法研究進展[J];色譜;2013年06期

6 楊建軍;秦環(huán)龍;;核不均一核糖核蛋白A1在腫瘤發(fā)病機制中作用的研究進展[J];胃腸病學(xué);2010年04期

7 謝奇;廖天安;謝莉莉;張智;邢孔才;趙秀麗;田亞光;;海南省12歲學(xué)生口腔健康狀況流行病學(xué)調(diào)查[J];中國學(xué)校衛(wèi)生;2008年09期

8 鐘婭;楊汴生;何健;彭永平;梁振山;;河南省12歲兒童口腔健康流行病學(xué)調(diào)查[J];中國學(xué)校衛(wèi)生;2008年12期

9 袁啟明;;激光技術(shù)在醫(yī)學(xué)領(lǐng)域的應(yīng)用與發(fā)展[J];現(xiàn)代醫(yī)學(xué)儀器與應(yīng)用;2007年05期

10 孫妍;;年輕恒牙的治療特點[J];中國醫(yī)藥導(dǎo)報;2010年14期

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