cyclin G2荧光蛋白重组质粒的构建及其对宫颈癌HeLa细胞的作用

Construction of fluorescent protein recombinant plasmid with cyclin G2 and its function in HeLa cells

目的构建包含人cyclin G2基因的荧光蛋白重组质粒,探讨cyclin G2对宫颈癌He-La细胞凋亡的影响。方法运用RT-PCR和基因克隆技术构建pDsRed2-Cyclin G2重组质粒;脂质体法转染重组质粒至HeLa细胞中,荧光显微镜下观察其蛋白的表达和定位;流式细胞术检测HeLa细胞的凋亡率。结果成功地构建了pDsRed2-Cyclin G2荧光蛋白重组质粒,其融合蛋白在HeLa细胞中的定位为弥漫分布在细胞核和细胞质。转染重组质粒能够促进HeLa细胞的凋亡。结论外源性cy-clin G2转染至HeLa细胞中可促进其凋亡,抑制细胞增殖。

Objective To construct human cyclin G2 gene eukaryotic vector and study the effect on HeLa cells growth in vitro. Methods The pDsRed2-cyclin G2 fusion protein expression vectors were constructed by RT-PCR and gene cloning techniques. Then it was transferred into HeLa cells by cation lipofectin reagent. Expression, intracellular localization and function of this fusion protein in HeLa cells were observed by fluorescence microscope after transfection. The apoptosis were analyzed by FCM after 48 hours. Results cyclin G2 gene eukaryotic expression vector was constructed successfully, fusion proteins located diffusely in nucleus and cytoplasm of HeLa cells. The pDsRed2-G2 fusion protein can advance apoptosis of HeLa cells. Conclusion cyclin G2 gene can inhibit the growth of HeLa cells.