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《China Biotechnology》 2014-09
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Effect of Silencing Nup88 Gene by RNA Interference on Growth and Invasion in Human Breast Cancer MCF-7 Cell

LI Yu-qiang;ZHU Zhi-tu;WANG Wei;LI Chen;XU Na;WANG Yu;LI Nan;SUN Hong-zhi;The First Affiliated Hospital of Liaoning Medcial Uinversity;  
Objective:Construct recombinant lentivirus NUP88- shRNA mediated by the carrier,by RNAi technology respectively to observe silence after NUP88 MCF- 7 proliferation,adhesion,invasion and metastasis of influence,looking for new targets for clinical gene therapy of breast cancer.Methods Build NUP88 Lentivirus vectors,packaging test after drop,to MOI cell transfection breast cancer MCF-7,using RT-PCR and Westernblot test each mRNA and protein expression in MCF-7 cell efficiency;Determined by MTT method and flow cytometry assay,after testing NUP88 gene is interference effects on MCF-7 cell proliferation and apoptosis;Cell invasion after experimental detection NUP88 gene is interference effects on MCF-7 aggressivity.Results Four groups of virus and a negative control group are building a successful,drop degrees are 4E + 8 Tu/mL;RT-RCR and Western-blot test,the results show that the NUP88-shRNA transfection of MCF-7 cell group NUP88 mRNA and protein expression and the negative transfection group and blank MCF- 7 cell group compared with significant difference statistically significant(P 0.01);Determination of NUP88-shRNA1 group of silence,the most efficient silence rate can reach 86%;Determined by MTT method,the results show that the experimental group after NUP88-shRNA1 Lentivirus transfection cell proliferation significantly reduced,compared with the blank group and the control group with significant difference(P0.05);Flow cytometry instrument detection of three groups of MCF- 7 cell apoptosis results show that the experimental group after lentivirus transfection cell apoptosis rate increased significantly,compared with the control group and blank group had significant difference(P 0.05);Cell invasion experiment show that the tumor cells to conventional training after 24 h,compared with the blank group and the negative control group,experimental group in membrane cells decreased significantly,with significant difference(P0.05).Conclution NUP88 recombinant lentivirus can suppress the MCF- 7 by RNAi successful NUP88 gene expression,and can significantly inhibit the proliferation and invasion ability in the distance.
【Fund】: 辽宁省教育厅教学计划(L201145);; 辽宁省高等学校优秀人才支持计划(LJQ2011091)资助项目
【CateGory Index】: R737.9
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