Overexpression of PKA regulatory subunits in ovarian cancer cells

The regulatory subunit of cAMP-dependent protein kinase (PKA) exists in two isoforms, RI and RII, which distinguish the PKA isozymes, type I (PKA-I) and type II (PKA-II). Evidence obtained from a variety of different experimental approaches has shown that the relative levels of type I and type II PKA in cells can play a major role in determining the balance between cell growth and differentiation. RIα transfected cells exhibit hyper-proliferative growth and RIIβ transfected cells revert to a relatively quiescent state. Profiling by microarray revealed equally profound changes in gene expression between RIα, RIIβ, and parental OVCAR cells. Keywords: genetic modification In order to characterize the effect of PKA type I and type II regulatory subunits on gene transcription at a global level, the PKA regulatory subunit genes for RIα and RIIβ were stably transfected into cells of the ovarian cancer cell line (OVCAR8). Three independent samples form each of RIα, RIIβ stably transfected cell lines as well as from parental cells were harvested for RNA isolation and subsequent microarray analysis.

环腺苷酸依赖性蛋白激酶（cAMP-dependent protein kinase，简称PKA）的调节亚基存在RI和RII两种同工型，二者可区分I型（PKA-I）与II型（PKA-II）两种PKA同工酶。
多项不同实验手段获取的证据显示，细胞内I型与II型PKA的相对水平，在调控细胞生长与分化的平衡过程中发挥核心作用。
转染RIα的细胞呈现过度增殖的生长表型，而转染RIIβ的细胞则恢复至相对静止的状态。通过基因芯片（microarray）进行的表达谱分析表明，RIα、RIIβ转染细胞与亲本OVCAR细胞之间的基因表达均发生了极为显著的变化。
关键词：基因修饰（genetic modification）
为了在全局水平上表征PKA I型与II型调节亚基对基因转录的影响，研究人员将RIα与RIIβ的PKA调节亚基基因稳定转染至卵巢癌细胞系OVCAR8中。
分别从RIα、RIIβ稳定转染细胞系以及亲本细胞中各获取3份独立样本，用于RNA提取及后续的基因芯片分析。