The Influence Of The Expression Of MDR-1in Irradiation-induced Drug Resistance Cell CNE1/R Of Human Nasopharyngeal Squamous Cell Carcinoma By SiRNA Interfering PECAM-1

Objective The mechanism of X-ray irradiation-induced MDR is unclear. It’s hot point and key point in the research fields of whether tumor cells irradiated by X-ray can generate MDR and finding the key elements and proving the its mechanism. The premier experimental results indicated PECAM-1is relative to the generation of MDR in nasopharyngeal Squamous cell carcinoma drug resistance cell CNE1/R induced by X-ray radiation. In this research, we interfered PECAM-1in CNE1/R cell, and detected the expression of MDR-1and study the relationship between PECAM-1and MDR-1.Methods Using RNA interference technology, designing PECAM-1’s siRNA fragments, and cloning into plasmid pSilencer2.1to build the recombinant plasmid pSilencer2.1-PECAM-1. pSilencer2.1and pSilencer2.1-PECAM-1were transfected into CNE1/R by Lipofectamine, namely the control group and the experimental group, respectively. Transfected cells were screened by G418, picked the cell monoclonal and expanded cultivation to obtain monoclonal cell lines. The total RNA and total protein were extracted in two types of cells to detect the expression of PECAM-1and MDR-1using Real time-PCR and Western blot after transfection.Results We had established the recombinant plasmid pSilencer2.1-PECAM-1. The results showed that the gene and protein expression of PECAM-1in the experimental group decreased by5.27times and3.40times, respectively, compared with the control group using Real time-PCR and Western blot, which suggested that we established the pSilencer2.1-PECAM-1/CNE1/R cell line, stably interfered the expression of PECAM-1. The gene and protein expression of MDR-1decreased by2.43times and2.79times, respectively, compared with the control group, namely with the decreasing of the expression of PECAM-1, the expression of MDR-1decreased, as well.Conclusion As the experiment results above indicated that the recombinant plasmid pSilencer2.1-PECAM-1and the pSilencer2.1-PECAM-1/CNE1/R cell line, stably interfered the expression of PECAM-1, were established. PECAM-1may be involved in the expression of MDR-1and the formation of radiation-induced multidrug resistance mechanisms.