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The Effects of shRNA Mediated Gene Silencing of Transcription Factor SNAI1 on Biological Phenotypes in Breast Cancer Cell Line MCF-7

Author: YuLiNa
Tutor: QuanChengShi
School: Jilin University
Course: Pathology and Pathophysiology
Keywords: Breast Cancer Transcription factor SNAI1 Gene silencing Biological phenotypes
CLC: R737.9
Type: Master's thesis
Year: 2010
Downloads: 104
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Abstract


Objective: in vitro cultured human breast cancer MCF-7 cells for the study, the study transcription factor the snail (SNAI1) gene silencing of MCF-7 cell biology phenotype . : SNAI1 gene sequence shRNA expressing green fluorescent protein ( Green Fluorescent Protein, GFP ) eukaryotic expression vector pGCsilencerTMU6/Neo/GFP in connection transfected MCF-7 cells and G418 selection to obtain plasmid relatively stable expression cells ; mRNA levels by RT-PCR the identification SNAI1 gene silencing effect ; the immunofluorescence detection SNAI1 gene protein levels of silence effect change and express positioning ; inverted microscope changes in cell morphology was observed ; cell count assay of cell proliferation ; cells zoned mark method detecting cells in vitro migration capacity ; Boyden chamber method detects cells in the in vitro invasion ability ; RT-PCR detection of epithelial markers claudin - 4 of claudin - 6 of claudin - 7 , of E-cadherin and between qualitative marker expression of MMP - 2 expression the level of change . Results the : SNAI1 gene silencing group relative to the control group, cell morphology change ; proliferation, and migration and invasion of diminished capacity ; cells of epithelial markers claudin-4, claudin-7, E-cadherin expression was elevated mesenchymal markers MMP 2 Expression reduced. Conclusion : SNAI1 gene silencing can inhibit MCF-7 cells malignant phenotype , and reverse the EMT process through the regulation of target genes .

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CLC: > Medicine, health > Oncology > Genitourinary tumors > Breast tumor
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