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RNA干擾TXR1基因?qū)Ρ茄拾┘毎退幍挠绊?/H1>
發(fā)布時間:2018-06-22 07:54

  本文選題:TXR1 + RNA干擾 ; 參考:《中南大學》2013年博士論文


【摘要】:目的:探索TXR1基因在鼻咽癌紫杉醇耐藥中的作用,利用RNA干擾技術(shù)抑制鼻咽癌紫杉醇耐藥細胞CNE-1/Taxol的TXR1基因表達,同時研究TXR1基因的表達與鼻咽癌紫杉醇耐藥的相關(guān)性。 方法: 1.通過脂質(zhì)體介導轉(zhuǎn)染siRNA至鼻咽癌紫杉醇耐藥細胞株CNE-1/Taxol。 2.利用實時熒光定量RT-PCR. Western-blot方法分別檢測RNA干擾前、后的鼻咽癌紫杉醇耐藥CNE-1/Taxol細胞株的TXR1基因表達差異。 3.利用MTT實驗檢測TXR1基因表達抑制后的鼻咽癌紫杉醇耐藥細胞CNE-1/Taxol的耐藥性變化,觀察TXR1基因在紫杉醇耐藥過程中的作用。 結(jié)果: 1.針對TXR1基因的siRNA轉(zhuǎn)染耐藥細胞CNE-1/Taxol后,耐藥細胞TXR1的mRNA及蛋白表達均明顯下調(diào)。干擾組和陰性對照組的mRNA表達分別是脂質(zhì)體組mRNA表達量的0.12±0.02倍、0.97±0.07倍,干擾組分別和陰性對照組、脂質(zhì)體組比較,差異具有統(tǒng)計學意義(P0.01)。干擾組、陰性對照組的蛋白表達量分別是脂質(zhì)體組的0.36±0.07、1.02±0.03倍,干擾組與脂質(zhì)體組、陰性對照組的蛋白表達相比較,差異有統(tǒng)計學意義(P0.01)。 2.TXRl基因表達抑制后,CNE-1/Taxol對紫杉醇的敏感性顯著增高,其IC50值降低了2.12倍,從而部分逆轉(zhuǎn)CNE-1/Taxol耐藥。 結(jié)論: 1.特異性siRNA分子能有效的抑制鼻咽癌紫杉醇耐藥細胞的TXR1基因表達。 2.運用RNA干擾技術(shù)抑制TXR1基因可能部分逆轉(zhuǎn)人鼻咽癌紫杉醇耐藥細胞CNE-1/Taxol的耐藥性。TXR1基因可能成為逆轉(zhuǎn)鼻咽癌細胞紫杉醇耐藥及腫瘤治療的靶分子。 圖7幅,表2個,參考文獻53篇
[Abstract]:Aim: to investigate the role of TXR1 gene in the resistance of nasopharyngeal carcinoma (NPC) to paclitaxel, and to investigate the relationship between TXR1 gene expression and taxol resistance in nasopharyngeal carcinoma (NPC) cells CNE-1 / Taxol by RNA interference. Methods: 1. Transfection of siRNA into CNE-1 / Taxol-2. 2 by liposome mediated transfection of siRNA into CNE-1 / Taxol- resistant nasopharyngeal carcinoma cell line. Real-time fluorescence quantitative RT-PCR was used. Western-blot method was used to detect the difference of TXR1 gene expression in CNE-1 / Taxol cell line before and after RNA interference. The drug resistance of CNE-1 / Taxol cells after TXR1 gene expression inhibition was detected by MTT assay, and the role of TXR1 gene in taxol resistance was observed. Results: 1. After transfection of siRNA targeting TXR1 gene into CNE-1 / Taxol, the mRNA and protein expression of TXR1 was down-regulated. The mRNA expression of interference group and negative control group was 0.12 鹵0.02 times 0.97 鹵0.07 times of that of liposome group, respectively. The difference between interference group and negative control group was statistically significant (P0.01). The protein expression in interference group and negative control group was 0.36 鹵0.07 鹵1.02 鹵0.03 times of that in liposome group, and the protein expression in interference group was higher than that in liposome group and negative control group. 2. The sensitivity of CNE-1 / Taxol to paclitaxel was significantly increased after the inhibition of TXRl gene expression, and the IC50 value of CNE-1 / Taxol decreased 2.12 times, which partially reversed the resistance of CNE-1 / Taxol. Conclusion: 1. Specific siRNA molecules can effectively inhibit the expression of TXR1 gene in paclitaxel-resistant nasopharyngeal carcinoma cells. 2. Inhibition of TXR1 gene by RNA interference may partially reverse the drug resistance of CNE-1 / Taxol cell line CNE-1 / Taxol. TXR1 gene may be a target molecule for reversing taxol resistance and tumor therapy of nasopharyngeal carcinoma cell line CNE-1 / Taxol. 7 figures, 2 tables, 53 references
【學位授予單位】:中南大學
【學位級別】:博士
【學位授予年份】:2013
【分類號】:R739.63

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