HOXD8基因在人卵巢癌細(xì)胞株SKOV3和SKOV3-DDP中的表達(dá)及卵巢癌順鉑耐藥的機(jī)制研究
發(fā)布時(shí)間:2018-04-04 23:18
本文選題:HOXD8 切入點(diǎn):卵巢癌 出處:《福建醫(yī)科大學(xué)》2014年碩士論文
【摘要】:目的:檢測(cè)同源異型盒基因D8(HOXD8)在人卵巢癌細(xì)胞株SKOV3和SKOV3-DDP中的表達(dá),探討卵巢癌順鉑耐藥的機(jī)制。 方法:體外培養(yǎng)人卵巢癌細(xì)胞株SKOV3及其配對(duì)順鉑耐藥細(xì)胞株SKOV3-DDP,應(yīng)用XTT比色實(shí)驗(yàn)檢測(cè)SKOV3、SKOV3-DDP細(xì)胞株順鉑處理后的半數(shù)抑制濃度(IC50),并計(jì)算耐藥指數(shù);半定量RT-PCR及實(shí)時(shí)熒光定量PCR、Elisa方法檢測(cè)HOXD8在SKOV3、SKOV3-DDP細(xì)胞株中的表達(dá),探討同源異型盒基因D8與卵巢癌順鉑耐藥的關(guān)系。 結(jié)果: 1.細(xì)胞經(jīng)不同濃度順鉑處理72小時(shí)后,隨著順鉑濃度的增加,細(xì)胞抑制率增高,顯示出明顯的劑量依賴(lài)關(guān)系;耐藥細(xì)胞株經(jīng)順鉑作用后,細(xì)胞抑制率明顯低于敏感株細(xì)胞株(p0.05)。XTT比色實(shí)驗(yàn)結(jié)果顯示,SKOV3細(xì)胞株順鉑IC50值為2.462±0.81ug/ml,SKOV3-DDP細(xì)胞株順鉑IC50值為9.174±0.24ug/ml,SKOV3-DDP細(xì)胞株的耐藥指數(shù)為3.73,可進(jìn)行耐藥實(shí)驗(yàn)。 2.半定量RT-PCR實(shí)驗(yàn)結(jié)果顯示,SKOV3細(xì)胞株中HOXD8mRNA的表達(dá)水平為0.238±0.037,SKOV3-DDP細(xì)胞株中HOXD8mRNA的表達(dá)水平為0.707±0.026。與SKOV3細(xì)胞株比較,SKOV3-DDP細(xì)胞株中HOXD8mRNA的相對(duì)表達(dá)量明顯升高,差異有統(tǒng)計(jì)學(xué)意義(p0.05)。熒光定量PCR實(shí)驗(yàn)結(jié)果顯示,SKOV3細(xì)胞株中HOXD8mRNA的表達(dá)水平為0.1711±0.026,SKOV3-DDP細(xì)胞株中HOXD8mRNA的表達(dá)水平為0.5761±0.037。與SKOV3細(xì)胞株比較,SKOV3-DDP細(xì)胞株中HOXD8mRNA的相對(duì)表達(dá)量明顯升高,差異有統(tǒng)計(jì)學(xué)意義(p0.05)。 3. Elisa實(shí)驗(yàn)結(jié)果顯示,SKOV3細(xì)胞株中HOXD8蛋白的表達(dá)水平為1584.4±2.08pg/ml, SKOV3-DDP細(xì)胞株中HOXD8蛋白的表達(dá)水平為4423.3±1.52pg/ml。與SKOV3細(xì)胞株比較,SKOV3-DDP細(xì)胞株中HOXD8蛋白的表達(dá)量明顯升高,差異有統(tǒng)計(jì)學(xué)意義(p0.05)。 結(jié)論: HOXD8mRNA與HOXD8蛋白在體外培養(yǎng)的卵巢癌細(xì)胞株SKOV3及其順鉑耐藥株SKOV3-DDP及中均有表達(dá)。 SKOV3-DDP細(xì)胞株中HOXD8的表達(dá)明顯高于其在SKOV3細(xì)胞株中的表達(dá),其表達(dá)的升高與卵巢癌細(xì)胞順鉑耐藥存在明確的相關(guān)性。
[Abstract]:Aim: to detect the expression of homologous cassette gene D8 (HOXD8) in human ovarian cancer cell line SKOV3 and SKOV3-DDP, and to explore the mechanism of cisplatin resistance in ovarian cancer.Methods: human ovarian cancer cell line SKOV3 and its cisplatin resistant cell line SKOV3-DDP were cultured in vitro. The IC50 inhibition concentration (IC50) was measured by XTT colorimetric assay after treatment with cisplatin, and the drug resistance index was calculated.Semi-quantitative RT-PCR and real-time fluorescence quantitative PCR Elisa were used to detect the expression of HOXD8 in SKOV3- SKOV3-DDP cell line, and to explore the relationship between homologous cassette gene D8 and cisplatin resistance in ovarian cancer.Results:1.After treated with different concentrations of cisplatin for 72 hours, the cell inhibition rate increased with the increase of cisplatin concentration, which showed a significant dose-dependent relationship.緇嗚優(yōu)鎶戝埗鐜囨槑鏄句綆浜庢晱鎰熸牚緇嗚優(yōu)鏍,
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