Real-time detection of gene expression in cancer cells using molecular beacon imaging: new strategies for cancer research.
Development of novel approaches for quantitative analysis of gene expression in intact tumor cells should provide new means for cancer detection and for studying the response of cancer cells to biological and therapeutic reagents. We developed procedures for detecting the levels of expression of multiple genes in fixed as well as viable cells using molecular beacon imaging technology. We found that simultaneous delivery of molecular beacons targeting survivin and cyclin D1 mRNAs produced strong fluorescence in breast cancer but not in normal breast cells. Importantly, fluorescence intensity correlated well with the level of gene expression in the cells detected by real-time reverse transcription-PCR or Western blot analysis. We further show that molecular beacons can detect changes of survivin gene expression in viable cancer cells following epidermal growth factor stimulation, docetaxel treatment, and overexpression of p53 gene. Thus, molecular beacon imaging is a simple and specific method for detecting gene expression in cancer cells. It has great potential for cancer detection and drug development.
Peng
Xiang-Hong
XH
Department of Surgery, Winship Cancer Institute, Emory University School of Medicine, Atlanta, Georgia 30322, USA.
Cao
Ze-Hong
ZH
Xia
Jin-Tang
JT
Carlson
Grant W
GW
Lewis
Melinda M
MM
Wood
William C
WC
Yang
Lily
L
D001483:Base Sequence; D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism*; D015153:Blotting, Western; D001940:Breast / Q000378:metabolism / Q000473:pathology; D001943:Breast Neoplasms / Q000175:diagnosis* / Q000235:genetics / Q000378:metabolism; D005260:Female; D005456:Fluorescent Dyes / Q000378:metabolism; D006801:Humans; D007090:Image Interpretation, Computer-Assisted; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism*; D015335:Molecular Probes; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D012689:Sequence Homology, Nucleic Acid; D000077022:Survivin; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism*
C107489:BIRC5 protein, human; D014408:Biomarkers, Tumor; D005456:Fluorescent Dyes; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins; D015335:Molecular Probes; D009363:Neoplasm Proteins; D012333:RNA, Messenger; D000077022:Survivin; D016159:Tumor Suppressor Protein p53