Short hairpin RNA silencing the expression of insulin-like growth factor binding protein 3 in corpus cavernosal smooth muscle cell in rat

• DOI: 10.3760/cma.j.issn.1001-9030.2017.08.033
• Publication Date: 2017-08-08
• Authors: Pu Xiaoyong; Xiangxue Zhou; Hengjun Xiao; Chuqi Lin; Wei-Hua Peng; Xue‐cheng Bi; Dong Li; Jiumin Liu

Objective To screen the most effective short hairpin RNA (shRNA) which down-regulate insulin-like growth factor binding protein 3 (IGFBP-3) in corpus cavernosal smooth muscle cell (CCSMC) vitro.

Methods 4 pair IGFBP-3 small interfering (siRNA), and plasmid constructs for U6 promoter vector (pGPU6/GFP/Neo-IGFBP-3-shRNA) negative control were constructed. Corpus cells primary cultured divided into 6 groups: CCSMCs-only, transfection pGPU6/GFP/Neo-IGFBP-3 (611) group, (526) (866) (710) group (transfection with pGPU6/GFP/Neo-shNC). The silencing effection was determined by real-time quantitative polymerase chain reaction (Real-time PCR) Western blotting.

Results CCSMCs transfected shRNA showed an obviously decrease of mRNA levels compared CCSMCs CCSMCs-only group. inhibiting observed trasnfection pGPU6/GFP/Neo-shIGFBP3-526 72% 79% PCR blotting.

Conclusion shRNA can silence expression CCSMCs. It may be methods to improved erection vivo.

Key words: Erectile dysfunction; Insulin-like 3; Gene Therapy; siRNA Technology; Rats