|
| Status |
Public on Mar 27, 2007 |
| Title |
HT-29 cell line - shNOX1 - Rep 4 |
| Sample type |
RNA |
| |
|
| Source name |
HT-29 stable clone 6A4 (shNOX1)
|
| Organism |
Homo sapiens |
| Characteristics |
HT-29 colon cancer cell line
stably transfected with shNOX1
clone 6A4
|
| Extracted molecule |
total RNA |
| Label |
biotin
|
| |
|
| Description |
We have found that NOX1 over-expression in colon cancer cell lines as well as surgical samples suggests a role in colon cancer. We identified unique target sequences of siRNAs for post-transcriptional silencing of NOX1. Gene specific and scrambled (control) sequences carrying the U6 promoter/shRNA cassettes were cloned into a GFP expression vector. Stable clones were selected based on NOX1 expression after transient transfection of HT-29 cells. HT-29 parental cells, clone SA (scrambled) and clone 6A (gene specific) were utilized to establish xenografts in athymic mice to investigate the effect of NOX1 silencing on tumor growth and gene expression in vivo. A significant decrease was measured in the volume of tumors formed. The stable clones and corresponding xenografts were analyzed on microarray. The microarray data was validated with real-time PCR and western-blot analysis demonstrating the inhibition of genes important in cell cycle regulation and angiogenesis. The results of this study show silencing NOX1 levels of generated ROS decreased, the tumor formation attenuated in vivo suggesting NOX1 may be a therapeutic target for colon cancer.
|
| Data processing |
The following were applied using the affy library in the R-Project Bioconductor statistical tools package: RMA background correction, pmonly probe-level correction, quantile normalization, median-polish summary method.
|
| |
|
| Submission date |
Mar 27, 2006 |
| Last update date |
Mar 29, 2006 |
| Contact name |
Agnes Juhasz |
| E-mail(s) |
ajuhasz@coh.org
|
| Organization name |
City of Hope Compr. Cancer Center
|
| Department |
Medical Oncology
|
| Street address |
1500 E. Duarte Rd
|
| City |
Duarte |
| State/province |
CA |
| ZIP/Postal code |
91010 |
| Country |
USA |
| |
|
| Platform ID |
GPL571 |
| Series (1) |
| GSE4561 |
Inhibition of NOX1 Gene Expression with shRNA in Human Colon Cancer |
|
