Identification of genes that regulate epithelial cell migration using an siRNA screening approach.
To provide a systematic analysis of genes that regulate epithelial cell migration, we performed a high throughput wound healing screen with MCF-10A breast epithelial cells, using siRNAs targeting 1,081 human genes encoding phosphatases, kinases and proteins predicted to influence cell migration and adhesion. The primary screen identified three categories of hits: those that accelerate, those that inhibit and those that impair migration with associated effects on cell proliferation or metabolism. Extensive validation of all the hits yielded 66 high confidence genes that, when downregulated, either accelerated or impaired migration; 42 of these high confidence genes have not been previously associated with motility or adhesion. Time-lapse video microscopy revealed a broad spectrum of phenotypic changes involving alterations in the extent and nature of disruption of cell-cell adhesion, directionality of motility, cell polarity and shape, and protrusion dynamics. Informatics analysis highlighted three major signalling nodes, beta-catenin, beta1-integrin and actin, and a large proportion of the genes that accelerated migration impaired cell-cell adhesion.
Simpson
Kaylene J
KJ
Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, USA.
Selfors
Laura M
LM
Bui
James
J
Reynolds
Angela
A
Leake
Devin
D
Khvorova
Anastasia
A
Brugge
Joan S
JS
D000199:Actins / Q000235:genetics / Q000378:metabolism; D015815:Cell Adhesion Molecules / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics*; D049109:Cell Proliferation; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism*; D053263:Gene Regulatory Networks; D005820:Genetic Testing; D006801:Humans; D019012:Integrin beta1 / Q000235:genetics / Q000378:metabolism; D010641:Phenotype; D034741:RNA, Small Interfering / Q000235:genetics*; D015203:Reproducibility of Results; D015398:Signal Transduction; D013997:Time Factors; D014162:Transfection; D014945:Wound Healing; D051176:beta Catenin / Q000235:genetics / Q000378:metabolism
D000199:Actins; D015815:Cell Adhesion Molecules; D019012:Integrin beta1; D034741:RNA, Small Interfering; D051176:beta Catenin