本文選題：宮頸腫瘤 + miR��； 參考：《中華腫瘤防治雜志》2015年14期

【摘要】：目的探討miR21基因表達(dá)改變對(duì)宮頸癌Siha細(xì)胞順鉑敏感性的影響。方法采用脂質(zhì)體介導(dǎo)法分別以micrOFFTMmiR21inhibitor、micrONTMmiR21mimic、miR21陰性對(duì)照試劑轉(zhuǎn)染Siha細(xì)胞,實(shí)驗(yàn)分為miR21下調(diào)組、miR21上調(diào)組、陰性對(duì)照組及空白對(duì)照組。實(shí)時(shí)熒光定量PCR檢測(cè)轉(zhuǎn)染后各組細(xì)胞中miR21基因的表達(dá)水平;CCK-8比色法檢測(cè)各組細(xì)胞對(duì)順鉑的半抑制率濃度(50%inhibitory concentration,IC50值);AnnexinⅤ/PI法檢測(cè)順鉑處理后各組細(xì)胞的凋亡率;實(shí)時(shí)熒光定量PCR和蛋白質(zhì)印跡法,分別從mRNA和蛋白水平檢測(cè)順鉑對(duì)各組細(xì)胞中PTEN、PDCD4基因表達(dá)的影響。結(jié)果實(shí)時(shí)熒光定量PCR檢測(cè)miR21基因的表達(dá)結(jié)果顯示,miR21下調(diào)組的表達(dá)明顯低于陰性對(duì)照組,miR21上調(diào)組的表達(dá)明顯高于陰性對(duì)照組,F=255.525,P0.001。CCK-8比色法檢測(cè)結(jié)果顯示,順鉑對(duì)miR21下調(diào)組的IC50值為(2.44±0.69)μg/mL,陰性對(duì)照組為(3.96±0.07)μg/mL;miR21上調(diào)組為(6.93±0.07)μg/mL,空白對(duì)照組為(4.05±0.03)μg/mL,F=870.118,P0.001。AnnexinⅤ/PI檢測(cè)結(jié)果顯示,5μg/mL順鉑誘導(dǎo)miR21下調(diào)組的凋亡率為(64.36±2.47)%,陰性對(duì)照組為(4.2±0.17)%;miR21上調(diào)組為(0.06±0.03)%,空白對(duì)照組為(4.14±0.25)%,χ2=10.385,P=0.016。實(shí)時(shí)熒光定量PCR檢測(cè)PTEN和PDCD4結(jié)果顯示,miR21下調(diào)組PTEN的mRNA表達(dá)相較于陰性對(duì)照組顯著增加,miR21上調(diào)組PTEN的mRNA表達(dá)相較于陰性對(duì)照組降低,F=174.057,P0.001;miR21下調(diào)組PDCD4的mRNA表達(dá)相較于陰性對(duì)照組顯著增加,miR21上調(diào)組PDCD4的mRNA表達(dá)相較于陰性對(duì)照組降低,F=491.283,P0.001。蛋白質(zhì)印跡法檢測(cè)結(jié)果顯示,miR21下調(diào)組PTEN蛋白表達(dá)量為8.845±0.062,陰性對(duì)照組7.695±0.056;miR21上調(diào)組為6.908±0.058,空白對(duì)照組為1,F=16.036,P0.01。miR21下調(diào)組PDCD4蛋白表達(dá)量為9.936±0.036,陰性對(duì)照組為8.728±0.019;miR21上調(diào)組為7.838±0.066,空白對(duì)照組為1,F=55.323,P0.001。結(jié)論下調(diào)miR21基因可能增加宮頸癌Siha細(xì)胞對(duì)順鉑的敏感性,上調(diào)miR21基因后可能減弱宮頸癌Siha細(xì)胞對(duì)順鉑的敏感性。
[Abstract]:Objective to investigate the effect of miR21 gene expression on cisplatin sensitivity in Siha cells. Methods Siha cells were transfected with micrOFFTMmiR21inhibitory or micrONTMmiR21mimicmiR21 negative control reagent by liposome-mediated method. The cells were divided into three groups: miR21 up-regulated group, negative control group and blank control group. The expression level of miR21 gene in the transfected cells was detected by real-time fluorescence quantitative PCR and the apoptosis rate of the cells treated with cisplatin was detected by CCK-8 colorimetric assay and the IC50 value of inhibitory concentration of Cisplatin was determined by Annexin 鈪，

本文編號(hào)：1988241