Alternate Splicing of CD44 Messenger RNA in Prostate Cancer Growth
Abstract
Loss of CD44 standard and increased splice variant form CD44v7-10 facilitate prostate cancer (PC) invasion. Mitogen-activated protein kinase (MAPK) pathways and paracrine calcitonin may dysregulate CD44. CD44 total and CD44v7-10 RNA or protein were assessed in androgen-independent PC with known high CD44v7-10 expression and in BPH-1 cells in response to exogenous calcitonin and to inhibitors of protein kinase A, MEK, JNK, or p38 kinase. Inhibition of MEK or p38 but not JNK reduced CD44 RNA in cancer and benign cells. Calcitonin, in calcitonin receptor-positive cells only, caused suppression of CD44 total but increase in variant, the latter apparently dependent on the p38 pathway. See the attached, submitted manuscript. Aim 2: In vivo trials of altering CD44 in mouse xenografts. We spent several months cloning our DNA sequences for CD44 standard overexpression and CD44 variant RNAi out of pTracer and into pAAV-IRES-GFP to use the latter plasmid to establish therapeutic adenoassociated virus (AAV). However, when C4-2 prostate cancer cells were infected, only 30% were GFP+ infected cells, and the low percent of viable cells, due to viral cytopathic effect, prohibited in vitro or in vivo use of AAV. We decided to revert to the pTracer approach with blasticidin cell selection. Matrigel invasion assay demonstrates that non-invaded cells have much higher GFP+ than invaded ones and by western blot analysis, non-invasion correlates with altered CD44 standard and variant.
Document Details
- Document Type
- Technical Report
- Publication Date
- Apr 01, 2008
- Accession Number
- ADA483727
Entities
People
- Kenneth A. Iczkowski
Organizations
- University of Colorado Health