|
| Status |
Public on Dec 23, 2006 |
| Title |
BxPC3 MSX2_vector |
| Sample type |
RNA |
| |
|
| Source name |
B7, MSX2-overexpressing BxPC3 with expression vectors
|
| Organism |
Homo sapiens |
| Characteristics |
Tissue:pancreatic cancer cell line
|
| Biomaterial provider |
American Type Culture Collection (ATCC, Manassas, VA)
|
| Treatment protocol |
Transfections of cells with expression vectors (MSX2 cDNA or vector alone) were performed using FuGENE 6 (Roche, Indianapolis, IN) as recommended by the supplier and cell lines were selected with 800ug/ml of G418 (Invitrogen).
|
| Growth protocol |
Grown in modified Eagle’s medium (MEM, Invitrogen, Grand Island, NY) containing 10% fetal bovine serum (Miles, Kankakee, IL), maintained at 37C in 5% CO2 in a humidified environment.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Procedures were performed according to the manufacturer (CodeLinkTM, whole Human Genome Expression Bioarray, Amersham Biosciences) ’s protocol.
|
| Label |
Cy5-streptavidin
|
| Label protocol |
Incubation with Cy5-streptavidin at room temperature for 30 min in the dark.
|
| |
|
| Hybridization protocol |
10ug aliquots of total RNA was faragmented at 94C for 20 min in the presence of magnesium. The fragmented RNA was hybridized to Uniset Human Whole Genome Expression Bioarray slides in hybridization buffer at 37C for 24 hours in an INNOVATM 4080 Shaking Incubator (New Brunswick Scientific, Edison, NJ) at 300rpm. After hybridization the arrays were washed in 0.75x TNT buffer at 46C for 1 hour followed by incubation with Cy5-streptavidin at room temperature for 30 min in the dark. Arrays were then washed in 1x TNT four times for 5 min each followed by a rinse in 0.1x SSC/0.05% Tween 20 in water. The slides were then dried by centrifugation and kept in the dark until scanning.
|
| Scan protocol |
Array slides were scanned using an Array worx (GE Healthcare Bio-Sciences Corp., Piscatway, NJ), and expression values were measured and manipulated subsequently by CodeLinkTM Expression Analysis v.4.0 software (Amersham Biosciences).
|
| Description |
Twist 1 expression was significantly associated with MSX2 expression in human pancreatic carcinomas and was down-regulated in MSX2 pancreatic cancer cells by transfected with a small interfering RNA expression vector to inactivated MSX2.
|
| Data processing |
Processed by CodeLinkTM Expression Analysis v.4.0 software (Amersham Biosciences).
|
| |
|
| Submission date |
Dec 20, 2006 |
| Last update date |
Dec 22, 2006 |
| Contact name |
Wataru Fujibuchi |
| E-mail(s) |
fujibuchi-wataru@aist.go.jp
|
| Organization name |
Advanced Industrial Science and Technology
|
| Department |
Computational Biology Research Center
|
| Street address |
2-42 Aomi, Koto-ku
|
| City |
Tokyo |
| State/province |
Tokyo |
| ZIP/Postal code |
135-0064 |
| Country |
Japan |
| |
|
| Platform ID |
GPL2895 |
| Series (1) |
| GSE6585 |
MSX2 enhances the malignant phenotype and associates with Twist 1 expression in human pancreatic cancer |
|
