Expression of Taxol Resistant Gene1in Taxol Resistant Cell Lines of Nasopharyngeal Carcinoma Cells
|School||Central South University|
|Course||Department of Otolaryngology,|
|Keywords||TXR1 nasopharyngeal carcinoma siRNA paclitaxel Q-PCR|
Objectives1. To study the relation between TXR1and paclitaxel-resistant by doing research on the TXR1expression of human nasopharyngeal carcinoma parental cells and the paclitaxel resisitant cells and investigating the correlation between TXR1and paclitaxel resistance index.2. To screen the most efficient siRNA for inhibiting the TXR1expression and do research on the reversal of nasopharyngeal carcinoma paclitaxel resistance.Methods1.Extract RNA and protein from human nasopharyngeal carcinoma parental cells and the paclitaxel resisitant cells and RTFQ-PCR,Western blot are used to detect the expression of TXR1in nasopharyngeal carcinoma parental cells from the level of mRNA and protein.2.Use lip2000carrier to transfect siRNA into the nasopharyngeal carcinoma paclitaxel resisitant cells, extract protein24hours later,and then evaluate the expression of TXR1from different siRNA depending on the extracting protein.Results1.TXR1is expressed both in nasopharyngeal carcinoma parental cells and the paclitaxel resisitant cells, and TXR1in paclitaxel resisitant cells is higher than in nasopharyngeal carcinoma parental cells, and its expression level was positively correlated to the degree of drug resistance (R2=0.979)2.The four TXR1gene siRNA (siRNATXR1-22TXR1-311, TXR1-654, Negative control FAM) were transfected into the nasopharyngeal carcinoma paclitaxel resisitant cells HNE-2/Taxol by lip2000carrier,extract protein24hours later,and then evaluate the expression of targeted TXR1from different siRNA depending on the extracting protein,the result shows the strongest inhibition is siRNA TXR1-22.3.Cells transfected by Negative control FAM is observed and counted under the inverted fluorescence microscope,and get a higher transfection rate (>85%),which indirectly tells that lip2000is a good carrier.Conclusion1.TXR1in paclitaxel resisitant cells is higher than in nasopharyngeal carcinoma parental cells,it prompts that there may be a relationship between TXR1and paclitaxel-resistant,which lays the foundation for the reversal of drug resistance2.On the protein level, siRNA TXR1-22is the strongest inhibition in all of the siRNA,which lays the foundation for the later experiments.