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Optomizing Transfection Efficiency of Cervical Cancer Cells Transfected by Cationic Liposomes LipofectamineTM2000

  • Huang, Fei (Department of Pathology, The Affiliated Tumor Hospital Of Xinjiang Medical University) ;
  • Zhao, Feng (Department of Pathology, The Affiliated Tumor Hospital Of Xinjiang Medical University) ;
  • Liang, Li-Ping (Department of Pathology, The Affiliated Tumor Hospital Of Xinjiang Medical University) ;
  • Zhou, Mei (Department of Pathology, The Affiliated Tumor Hospital Of Xinjiang Medical University) ;
  • Qu, Zhi-Ling (Department of Pathology, Tongji Medical College, Huazhong University of Science and Technology) ;
  • Cao, Yan-Zhen (Department of Pathology, The Affiliated Tumor Hospital Of Xinjiang Medical University) ;
  • Lin, Chen (Department of Pathology, College of Basic Medicine, Xinjiang Medical University)
  • Published : 2015.12.03

Abstract

Background: Currently, cationic liposome has become the commonly used vehicles for gene transfection. Furthermore, one of the most significant steps in microRNAs expression studies is transferring microRNAs into cell cultures successfully. In this study we aim to approach the feasibility of transfection of cervical cancer cell lines mediated by liposome and to obtain the optimized transfection condition for cervical cancer cell lines. Materials and Methods: $Lipofectamine^{TM}2000$ as the carrier, miR-101 mimic was transfected into Hela cells and Siha cells. Using green fluorescent protein as reporter gene, to set different groups according to cell seeding density, the amount of miRNA, miRNA and the proportion of Liposomes, Whether to add serum into medium to study their impact on the liposomal transfection efficiency. Finally, MTT assay was used to analyze the relative minimal cell toxicity of liposome reagents. Results: The seeding density of Hela cell line and Siha are $1.5{\times}10^4$ (per well of 24 well plates), miRNA amount is 1ul of both, the ratio of miRNA and liposome is 1:0.5 of Hela cell line; 1:0.7 of Siha cell line respectively, after 24 hours we can get the highest transfection efficiency. Compared with serum medium, only Siha cells cultured with serum-free medium obtained higher transfection efficiency before transfection (P<0.01). MTT assay showed that according to the above conditions which has the lowest cytotoxicity. Conclusions: The method of Liposome to transfected is a suitable way and it can be an efficient reagent for miRNA delivery for Hela cells and Siha cells in vitro. It may serve as a reference for the further research or application.

Keywords

MicroRNA;cervical cancer cell;cell transfection liposome;transfection efficiency

Acknowledgement

Supported by : Xinjiang Uygur Autonomous Region Natual Science Foundation

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