本文選題：喉腫瘤 + 鱗狀細胞癌�。� 參考：《山西醫(yī)科大學》2012年碩士論文

【摘要】：目的: 探討miR-210及其靶基因E2F3在喉鱗狀細胞癌細胞系中的表達，并證明兩者直接調控關系。同時探討miR-210及E2F3的表達與喉鱗癌臨床病理參數的關系，為喉鱗狀細胞癌的診斷和預后評估尋找輔助生物學指標奠定基礎。 方法: 1.通過qRT-PCR方法測定兩種喉癌細胞株中miR-210及其下游靶基因E2F3的表達水平，并通過雙熒光檢測驗證miR-210與E2F3之間的直接調控關系；2.通過qRT-PCR方法檢測20例喉鱗狀細胞癌和癌旁正常切緣新鮮樣本中miR-210和E2F3 mRNA水平，分析二者在喉鱗狀細胞癌中表達的相關性以及二者與喉鱗狀細胞癌患者各項臨床病理學參數的關系；3.采用免疫組化技術檢測喉鱗狀細胞癌組織及癌旁正常切緣組織石蠟標本中E2F3蛋白表達，探討E2F3在喉鱗狀細胞癌發(fā)生發(fā)展過程中可能的作用；4.生存率統計采用壽命表法和Kaplan-Meier法，其差異性檢驗用Log-rank法；5.采用Cox回歸模型分析E2F3及臨床病理因素在喉鱗癌患者預后中的價值。所有數據均采用SPSS 16.0軟件包進行統計學分析。 結果: 1.檢測兩種喉癌細胞株Hep-2和TU177中miR-210和E2F3水平，結果顯示：在miR-210相對高表達的Hep-2細胞株中E2F3表達水平較miR-210相對低表達的TU177細胞株中低，通過雙熒光檢測證實：miR-210+E2F3細胞株組較miR-210+mutE2F3細胞株組的熒光強度明顯減弱，且差異有統計學意義(P＜0.05)；miR-210+E2F3細胞株組較NC組的熒光強度明顯減弱，且差異有統計學意義(P＜0.05)； 2.統計學方法分析20例喉鱗狀細胞癌和癌旁正常切緣新鮮樣本中miR-210和E2F3的水平，結果顯示：miR-210在喉鱗狀細胞癌組織和癌旁正常切緣組織中表達水平的均數±標準差分別為15.488±9.371和4.437±3.039，二者差異有統計學意義(P=0.000)；E2F3mRNA在喉鱗狀細胞癌組織和癌旁正常切緣組織中表達水平的均數±標準差分別為6.683±5.599和2.728±1.431，，二者差異有統計學意義(P=0.008)；喉鱗狀細胞癌新鮮組織的miR-210和E2F3 mRNA表達水平之間呈線性負相關(rs=-0.645, P=0.002)； 3.免疫組化檢測E2F3蛋白在喉鱗狀細胞癌與癌旁正常切緣中的表達率，結果顯示：兩組中E2F3蛋白陽性表達率分別為90.44% (123/136)和27.85% (22/79)，差異具有顯著統計學差異(P＜0.001)，E2F3蛋白胞核中高表達與喉鱗狀細胞癌患者年齡、臨床分期、組織分化和腫瘤分型有關；胞漿中高表達只于腫瘤分化有關(P＜0.05)； 4.單因素分析顯示：臨床分期、淋巴結有無轉移、T分期、E2F3蛋白核表達和腫瘤分型是喉鱗狀細胞癌患者生存期的影響因素（P＜0.05）； 5.多因素分析顯示：T分期、淋巴結有無轉移和E2F3蛋白核表達是影響喉鱗狀細胞癌患者預后的危險因素（P＜0.05）。 結論: 1. miR-210與靶基因E2F3之間存在直接調控關系； 2. E2F3蛋白核表達可能參與喉鱗狀細胞癌發(fā)生發(fā)展過程； 3. T分期、淋巴結有無轉移和E2F3蛋白核表達是喉鱗狀細胞癌患者預后影響因素，檢測E2F3蛋白核表達可為判斷喉鱗狀細胞癌患者預后評估提供幫助。
[Abstract]:Purpose :
To investigate the expression of miR - 210 and its target gene E2F3 in laryngeal squamous cell carcinoma cell lines and to demonstrate their direct control relationship . Meanwhile , the relationship between expression of miR - 210 and E2F3 and the clinical pathological parameters of laryngeal squamous cell carcinoma was discussed , which laid the foundation for the diagnosis and prognosis evaluation of laryngeal squamous cell carcinoma .
Method :
1 . The expression level of miR - 210 and its downstream target gene E2F3 was determined by qRT - PCR , and the direct control relationship between miR - 210 and E2F3 was verified by double fluorescence detection ;
2 . The levels of miR - 210 and E2F3 mRNA were detected by qRT - PCR in the fresh samples of laryngeal squamous cell carcinoma and adjacent normal margins , and the correlation between the expression of both in laryngeal squamous cell carcinoma and the clinical pathological parameters of laryngeal squamous cell carcinoma were analyzed .
3 . The expression of E2F3 protein in paraffin specimens of laryngeal squamous cell carcinoma and normal margin tissue was detected by immunohistochemistry , and the possible role of E2F3 in the development of laryngeal squamous cell carcinoma was discussed .
4 . Life table method and Kaplan - Meier method were used for survival statistics . Log - rank method was used for differential test .
5 . Cox regression model was used to analyze the value of E2F3 and clinicopathological factors in the prognosis of laryngeal squamous cell carcinoma . All the data were analyzed by SPSS 16.0 software package .
Results :
1 . The levels of miR - 210 and E2F3 in the two types of laryngeal cancer cell lines were tested . The results showed that the expression level of E2F3 was lower than that of the low - expressed TU177 cell line of miR - 210 . The results showed that the fluorescence intensity of the miR - 210 + E2F3 cell line group was significantly lower than that of the miR - 210 + mutE2F3 cell line group , and the difference was statistically significant ( P < 0.05 ) .
Compared with NC group , the fluorescence intensity of the miR - 210 + E2F3 cell line group was significantly decreased , and the difference was statistically significant ( P < 0.05 ) .

2 . The levels of miR - 210 and E2F3 in the fresh samples of laryngeal squamous cell carcinoma and adjacent normal margins were analyzed by statistical methods . The results showed that the mean 鹵 standard deviation of miR - 210 was 15.488 鹵 9.371 and 4.437 鹵 3.39 , respectively , in the tissues of laryngeal squamous cell carcinoma and adjacent normal margin tissues ( P = 0.000 ) .
The mean 鹵 SD of E2F3mRNA was 6.683 鹵 5.599 and 2.728 鹵 1.431 in normal margin of laryngeal squamous cell carcinoma ( P = 0.008 ) .
There was a linear negative correlation between miR - 210 and E2F3 mRNA expression levels in fresh tissue of laryngeal squamous cell carcinoma ( rs = - 0.645 , P = 0.002 ) ;

3 . The expression rate of E2F3 protein in laryngeal squamous cell carcinoma and normal margin of carcinoma was detected by immunohistochemistry . The results showed that the positive rates of E2F3 protein were 90.44 % ( 123 / 136 ) and 27.85 % ( 22 / 79 ) in both groups , and the difference was statistically significant ( P < 0.001 ) .
High expression in cytoplasm was only related to tumor differentiation ( P < 0.05 ) .

4 . Single factor analysis showed that clinical stage , lymph node metastasis , T stage , E2F3 protein core expression and tumor typing were the influencing factors of survival time of laryngeal squamous cell carcinoma ( P < 0.05 ) ;

5 . Multi - factor analysis showed that T staging , lymph node metastasis and E2F3 protein expression were the risk factors affecting the prognosis of patients with laryngeal squamous cell carcinoma ( P < 0.05 ) .
Conclusion :
1.There is a direct regulatory relationship between miR - 210 and the target gene E2F3 ;

2 . The expression of E2F3 protein may be involved in the development of laryngeal squamous cell carcinoma .

3.T staging , lymph node metastasis and the expression of E2F3 protein were the prognostic factors in laryngeal squamous cell carcinoma .
【學位授予單位】：山西醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R739.65

【參考文獻】

相關期刊論文 前3條

1 李錦秀;李家兵;張盼;姚惠;;E2F3在前列腺癌中的表達及意義[J];現代泌尿外科雜志;2009年04期

2 王歡;林蓓;;卵巢上皮性腫瘤組織中核轉錄因子E2F3蛋白的表達及其臨床意義[J];現代腫瘤醫(yī)學;2009年05期

3 劉連新,姜洪池,朱安龍,王秀琴,周津,吳e

本文編號：2095544