TXR1基因在鼻咽癌紫杉醇耐药细胞系中的表达

发布时间：2018-03-11 14:22

本文选题：TXR1　切入点：鼻咽癌　出处：《中南大学》2012年硕士论文　论文类型：学位论文

【摘要】：目的 1.通过研究人类鼻咽癌亲本细胞与紫杉醇耐药细胞中TXR1的表达情况,及TXR1的表达与鼻咽癌紫杉醇耐药的相关性,探讨其与紫杉醇耐药的关系。 2.筛选高效抑制TXR1基因表达的siRNA,用于鼻咽癌紫杉醇耐药逆转的研究。方法 1.从鼻咽癌亲本及紫杉醇耐药细胞中提取RNA及蛋白质,用荧光定量RT-PCR、Western-blot方法分别在mRNA水平及蛋白水平来检测鼻咽癌细胞中TXR1的表达情况。 2.通过1ip2000载体将TXR1基因的siRNA导入鼻咽癌耐药细胞,24小时后提取的蛋白质,来检测不同siRNA对TXR1表达的影响。结果 1.TXR1在鼻咽癌亲本细胞、紫杉醇耐药细胞中均有表达,及耐药细胞中TXR1的含量比相对应亲本细胞中的含量高,且与耐药指数呈正相关(P2=0.979)。 2.四条TXR1基因的siRNA (siRNA TXR1-22、TXR1-311、 TXR1-654、Negative control FAM),通过lip2000分别转染鼻咽癌紫杉醇耐药细胞HNE-2/Taxol,转染24h后,提取蛋白,利用Western blot技术在蛋白水平检测不同siRNA对靶基因TXR1表达的影响,结果显示TXR1-22对靶基因的抑制作用最强。 3. Negative control FAM转染的细胞在倒置荧光显微镜下计数,得到较高的转染率(85%),间接说明lip2000转染效率高。结论 1.TXR1在紫杉醇耐药细胞中的含量高于鼻咽癌亲本细胞,说明TXR1可能与紫杉醇耐药有关,并为临床上逆转紫杉醇耐药奠定了基础。 2.在蛋白质水平,siRNA TXR1-22是所有设计的siRN中对TXR1抑制最强,这为后续的实验奠定了基础。
[Abstract]:Purpose. 1. To investigate the expression of TXR1 in human nasopharyngeal carcinoma (NPC) parental cells and paclitaxel resistant cells, and the relationship between TXR1 expression and paclitaxel resistance. 2. SiRNAs which can inhibit the expression of TXR1 gene in nasopharyngeal carcinoma were screened and used to reverse paclitaxel resistance in nasopharyngeal carcinoma. Method. 1. RNA and protein were extracted from nasopharyngeal carcinoma (NPC) parents and paclitaxel resistant cells. The expression of TXR1 in nasopharyngeal carcinoma cells was detected by RT-PCR Western-blot at mRNA level and protein level, respectively. 2. The siRNA of TXR1 gene was introduced into the drug-resistant nasopharyngeal carcinoma cells 24 hours later by 1ip2000 vector to detect the effect of different siRNA on the expression of TXR1. Results. 1. TXR1 was expressed in the parent cells of nasopharyngeal carcinoma and paclitaxel resistant cells, and the content of TXR1 in the resistant cells was higher than that in the corresponding parent cells, and there was a positive correlation between TXR1 and drug resistance index. 2.The siRNA siRNA TXR1-22nTXR1-311 and TXR1-654Negative control FAMN of four TXR1 genes were transfected into nasopharyngeal carcinoma paclitaxel resistant cell line HNE-2 / Taxol by lip2000 respectively. After 24 hours of transfection, the protein was extracted, and the effect of different siRNA on TXR1 expression of target gene was detected at protein level by Western blot technique. The results showed that TXR1-22 had the strongest inhibitory effect on target gene. 3.The cells transfected with Negative control FAM were counted under inverted fluorescence microscope and the transfection efficiency was 85%, which indirectly indicated that the transfection efficiency of lip2000 was high. Conclusion. 1. The content of TXR1 in paclitaxel resistant cells was higher than that in nasopharyngeal carcinoma parent cells, which suggested that TXR1 might be related to paclitaxel resistance and laid a foundation for clinical reversal of taxol resistance. 2. SiRNA TXR1-22 is the most potent inhibitor of TXR1 among all the designed siRN at protein level, which lays the foundation for further experiments.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R739.63

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