siRNA targeting Her 2 retroviral vector and its expression in the SKOV3.docVIP

 PAGE \* MERGEFORMAT 21 siRNA targeting Her 2 retroviral vector and its expression in the SKOV3 Authors: Zhou, Bin Ling, high-ting, Chen Min, Feng Ting-qing, Cheng Zhixiang, Yuan-Yuan Zhu, Wei-Dong Zhao, SHI Yong-yun, Wang Mei-Mei, ZHU Huai-ping 【Abstract】 Objective To construct and identify Her  2-specific siRNA retroviral vector to import SKOV3 ovarian cancer cells, and preliminary screening effectively inhibited Her  2 expression in cell lines. Methods for in vitro synthesized with Her  2-specific oligonucleotide sequences and cloned into the retroviral vector RNAi  Ready pSIREN  RetroQ, the carrier construct by sequencing, restriction enzyme digestion, the liposome-mediated transfer transfected into the virus packaging cell line PT67 in puromycin selection, and selection of cell clones, harvesting virus supernatant, the virus supernatant transfected SKOV3, and after puromycin selection to ensure a stable suppression Her  2 expression in SKOV3 cell lines, and respectively, through RT  PCR and immunohistochemistry were used to identify Her  2 expression inhibitory effect. The result successfully constructed recombinant retroviral vector, the virus packaging cell line transfected with viral supernatant obtained successfully transfected SKOV3, and inhibit Her  2 expression. Conclusion inhibit Her  2 expression in SKOV3 cell line, in order to observe the transfected retroviral siRNA expression vector of changes in the biological behavior of malignant tumor cells has laid a experimental basis. Keywords: Her  2; RNA interference; retroviral vector Abstract: Objective To establish the stable SKOV3 cell lines with a persistent knockdown of Her  2 by transfecting the recombinant retroviral vectors of siRNA specific for Her  2 into SKOV3 cell line. Methods We synthesized oligonucleotides for Her  2 in vitro, and cloned them into retroviral vector RNAi  Ready pSIREN  Retro  Q. Subsequently the plasmids were sequenced and di