本文選題：喉腫瘤 + 鱗狀細(xì)胞癌��； 參考：《山西醫(yī)科大學(xué)》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(xì)胞癌細(xì)胞系中的表達(dá)，并證明兩者直接調(diào)控關(guān)系。同時(shí)探討miR-210及E2F3的表達(dá)與喉鱗癌臨床病理參數(shù)的關(guān)系，為喉鱗狀細(xì)胞癌的診斷和預(yù)后評(píng)估尋找輔助生物學(xué)指標(biāo)奠定基礎(chǔ)。 方法: 1.通過(guò)qRT-PCR方法測(cè)定兩種喉癌細(xì)胞株中miR-210及其下游靶基因E2F3的表達(dá)水平，并通過(guò)雙熒光檢測(cè)驗(yàn)證miR-210與E2F3之間的直接調(diào)控關(guān)系；2.通過(guò)qRT-PCR方法檢測(cè)20例喉鱗狀細(xì)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(xì)胞癌中表達(dá)的相關(guān)性以及二者與喉鱗狀細(xì)胞癌患者各項(xiàng)臨床病理學(xué)參數(shù)的關(guān)系；3.采用免疫組化技術(shù)檢測(cè)喉鱗狀細(xì)胞癌組織及癌旁正常切緣組織石蠟標(biāo)本中E2F3蛋白表達(dá)，探討E2F3在喉鱗狀細(xì)胞癌發(fā)生發(fā)展過(guò)程中可能的作用；4.生存率統(tǒng)計(jì)采用壽命表法和Kaplan-Meier法，其差異性檢驗(yàn)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(yù)后中的價(jià)值。所有數(shù)據(jù)均采用SPSS 16.0軟件包進(jìn)行統(tǒng)計(jì)學(xué)分析。 結(jié)果: 1.檢測(cè)兩種喉癌細(xì)胞株Hep-2和TU177中miR-210和E2F3水平，結(jié)果顯示：在miR-210相對(duì)高表達(dá)的Hep-2細(xì)胞株中E2F3表達(dá)水平較miR-210相對(duì)低表達(dá)的TU177細(xì)胞株中低，通過(guò)雙熒光檢測(cè)證實(shí)：miR-210+E2F3細(xì)胞株組較miR-210+mutE2F3細(xì)胞株組的熒光強(qiáng)度明顯減弱，且差異有統(tǒng)計(jì)學(xué)意義(P＜0.05)；miR-210+E2F3細(xì)胞株組較NC組的熒光強(qiáng)度明顯減弱，且差異有統(tǒng)計(jì)學(xué)意義(P＜0.05)； 2.統(tǒng)計(jì)學(xué)方法分析20例喉鱗狀細(xì)胞癌和癌旁正常切緣新鮮樣本中miR-210和E2F3的水平，結(jié)果顯示：miR-210在喉鱗狀細(xì)胞癌組織和癌旁正常切緣組織中表達(dá)水平的均數(shù)±標(biāo)準(zhǔn)差分別為15.488±9.371和4.437±3.039，二者差異有統(tǒng)計(jì)學(xué)意義(P=0.000)；E2F3mRNA在喉鱗狀細(xì)胞癌組織和癌旁正常切緣組織中表達(dá)水平的均數(shù)±標(biāo)準(zhǔn)差分別為6.683±5.599和2.728±1.431，，二者差異有統(tǒng)計(jì)學(xué)意義(P=0.008)；喉鱗狀細(xì)胞癌新鮮組織的miR-210和E2F3 mRNA表達(dá)水平之間呈線性負(fù)相關(guān)(rs=-0.645, P=0.002)； 3.免疫組化檢測(cè)E2F3蛋白在喉鱗狀細(xì)胞癌與癌旁正常切緣中的表達(dá)率，結(jié)果顯示：兩組中E2F3蛋白陽(yáng)性表達(dá)率分別為90.44% (123/136)和27.85% (22/79)，差異具有顯著統(tǒng)計(jì)學(xué)差異(P＜0.001)，E2F3蛋白胞核中高表達(dá)與喉鱗狀細(xì)胞癌患者年齡、臨床分期、組織分化和腫瘤分型有關(guān)；胞漿中高表達(dá)只于腫瘤分化有關(guān)(P＜0.05)； 4.單因素分析顯示：臨床分期、淋巴結(jié)有無(wú)轉(zhuǎn)移、T分期、E2F3蛋白核表達(dá)和腫瘤分型是喉鱗狀細(xì)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(jié)有無(wú)轉(zhuǎn)移和E2F3蛋白核表達(dá)是影響喉鱗狀細(xì)胞癌患者預(yù)后的危險(xiǎn)因素（P＜0.05）。 結(jié)論: 1. miR-210與靶基因E2F3之間存在直接調(diào)控關(guān)系； 2. E2F3蛋白核表達(dá)可能參與喉鱗狀細(xì)胞癌發(fā)生發(fā)展過(guò)程； 3. T分期、淋巴結(jié)有無(wú)轉(zhuǎn)移和E2F3蛋白核表達(dá)是喉鱗狀細(xì)胞癌患者預(yù)后影響因素，檢測(cè)E2F3蛋白核表達(dá)可為判斷喉鱗狀細(xì)胞癌患者預(yù)后評(píng)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(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類(lèi)號(hào)】：R739.65

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相關(guān)期刊論文 前3條

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本文編號(hào)：2095544