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Transduction of HOXB7 Gene Impact on Invasion of SKBR-3 Breast Cancer Cell Line

Author: LiuJiaHui
Tutor: WangJiJian
School: Chongqing Medical University
Course: Surgery
Keywords: Breast Cancer HOXB-7 Invasive Matrix metalloproteinase Justice transfected
CLC: R737.9
Type: Master's thesis
Year: 2009
Downloads: 27
Quote: 0
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Abstract


Objective To study the human homeobox gene HOXB - the core fragment justice transfected SKBR-3 breast cancer cell lines affect its invasiveness . HOXB-7 is extracted from the breast cancer cell line MCF-7 gene core coding region (HOXB7-CDS) fragment construct containing HOXB7-CDS fragment the plasmid ( PIRES2 -EGFP - of HOXB7 ) . Using semi-quantitative RT - PCR , Western blot blotting methods were used to detect breast cancer cell lines SKBR-3 HOXB-7 gene and fibroblast growth factor ( bFGF ) mRNA and protein expression levels . Will be divided into three groups: control group , negative control group (transfected with empty plasmid PIRES2 -EGFP was ) , HOXB - 7 justice SKBR-3 cells transfected group ( transfected to build on plasmid PIRES2 - of HOXB7 ) . Fluorescence microscopy transfection efficiency , RT-PCR , Western blot detection after transfection HOXB-7 and bFGF mRNA and protein expression . The Transwell chamber detected three groups of cell invasion , athletic ability changes ; Gelatin zymography assay of two matrix metalloproteinase activity in cell supernatants and protein ; Western blot to detect cell changes in matrix metalloproteinase -2 (MMP-2) . RT- PCR , Western blot results : of HOXB7 gene and bFGF factor in breast cancer MCF-7 cells positive expression , no expression in breast cancer SKBR-3 cells transfected with a plasmid PIRES2-HOXB7 SKBR-3 cells , HOXB-7 gene expression of the cell ; chamber experiments and tends motion experiments after transfection, cells whose invasion force and tend sports ability were rising ; RT-PCR , gelatin enzyme spectrum , Western blot results show HOXB7 justice turn dyed , SKBR - 3 of MMP-2 expression no significant changes in the expression of MMP -9 enhanced . Conclusion HOXB7 core fragment justice transfection allows breast cancer SKBR-3 cells HOXB-7 gene and bFGF factor expression and enhanced the expression of MMP-9 , MMP-2 had no significant effect ; same time enhance its migration , invasion , Tip HOXB-7 may play a role in breast cancer cell invasion and metastasis .

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