Establishment and research of a biomechanical model of bladder outflow obstruction at single cell level in vitro / 第三军医大学学报

ABSTRACT

Objective To establish a brand new model of bladder outflow obstruction (BOO) at single cell level in vitro to offer a more stable and scientific experimental base. Methods Recurrent mechanical stretch generated by vacuum facility was applied to cells attached to flexible membrane of special culture plate, which led to 10%, 20% and 30% elongation of them. Immunocytochemistry was used to analyze the expression of ?-actin (sign of contractile phenotype) and RT-PCR was performed to detect the dynamic changes of mRNA expression of PCNA and Western blotting to protein expression of PCNA and cdk2 kinase. MTT assay was used to observe the changes of proliferation of cells. Results In the case of gradual elongation, the expression of ?-actin rose at first and fell afterwards; the mRNA expression of PCNA and protein expression of PCNA and cdk2 kinase rose gradually. Changes in 30% elongation group were greater than the control (P