185 Impact of RhoGDI Gene Transfection of Bladder Smooth Muscle Contractility in a Validated Ex-vivo Murine Model
JOURNAL OF SEXUAL MEDICINE(2019)
Abstract
Plasmid-based gene therapy is an intriguing option for treating malignant and bladder pathologies. The RhoA pathway is involved in bladder smooth muscle regulation, cancer invasion and metastasis. Rho GDP-dissociation inhibitor (RhoGDI) is an inhibitor of the RhoA pathway. We validated ex-vivo bladder gene transfer to facilitate assessment of gene targets for treating bladder pathology. Basic Local Alignment Search Tool was used to identify human RhoGDI coding sequences and to compare between rats and humans, which were cloned into a eCMV-based expression vector. NBTII rat bladder cancer cell lines were transfected using FuGENE (Promega, USA) and human protein expression and interaction with endogenous RhoA were tested using flow cytometry, immunofluorescence, and RNA expression analysis. Bladders were harvested from female Lewis Rats (∼250g) and sectioned and cultured for 72-hours following transfection with RhoGDI and FuGENE. Transfected bladder tissues were analyzed as described above. Non-transfected cultured bladder segments were analyzed using myography for viability and intact smooth muscle physiology in response to 120 mM KCl and 30uM carbachol.
MoreTranslated text
Key words
Gene Therapy
AI Read Science
Must-Reading Tree
Example
![](https://originalfileserver.aminer.cn/sys/aminer/pubs/mrt_preview.jpeg)
Generate MRT to find the research sequence of this paper
Chat Paper
Summary is being generated by the instructions you defined