睾丸组织高表达基因PRKACG对细胞增殖的影响

The effect of a testicular highly expression gene PRKACG on cell proliferation

目的研究睾丸组织高表达cAMP依赖蛋白激酶催化亚基γ(protein kinase,cAMP-dependent,catalytic,gamma,PRKACG)基因的功能,观察其对NF-κB信号通路的激活作用和促细胞增殖作用。方法采用双荧光素酶报告基因系统,利用体外培养的293T细胞,分析PRKACG对NF-κB信号通路的激活作用,并观察293T细胞的形态学,通过细胞计数和CCK8观察其对细胞生长和细胞增殖的影响。结果转染PRKACG的293T细胞相对于对照组转染PCMV-Sport 6的细胞体积变大、饱满、触角伸长;CCK8和细胞计数的结果显示,过表达PRKACG的293T的生长速度明显高于对照组Sport 6。结论 PRKACG能够激活293T细胞的NF-κB信号通路,并可促进细胞增殖,提示PRKACG可能是睾丸发育和精子发生过程中的一个功能基因。

Objective： To study the function of a testicular highly expression gene PRKACG （protein kinase, cAMP-dependent, catalytic, gamma）, and find out novel diagnosis and therapeutic targets for the treatment of male infertility. Methods： The activation effect of PRKACG in the NF-~B signaling pathways that are related to spermatogenesis was analyzed by using dual luciferase report system, and the effect of PRKACG on cell proliferation was further studied by morphological observation, cell counting and CCK8 assay. Results：The 293T cells present the changes containing volume fill-out, full, antenna elongation, connection large expanse mostly when PRKACG transfect in the cells. Cell counting and CCK8 assays showed that PRKACG expression enhances overall cell growth rate compared with Sport 6. Conclusions： PRKACG could activate NF-kB signaling pathway, and promote cell proliferation, indicating that PRKACG might be a novel functional gene in testicular development and spermatogenesis.