摘要
目的观察慢病毒介导shRNA沉默PTTG基因表达对人生长激素型垂体腺瘤细胞生长的影响。方法原代培养人生长激素型垂体腺瘤细胞,分为正常对照组、阴性对照组和实验组。实验组构建PTTG-shRNA的慢病毒表达载体并转染肿瘤细胞,阴性对照组转染空慢病毒载体,正常对照组细胞不转染。流式细胞仪分析细胞转染效率,RT-PCR和Western blot检测肿瘤细胞PTTG mRNA和蛋白的表达水平,MTT检测细胞增殖情况,流式细胞仪检测细胞周期变化。结果与正常对照组和阴性对照组比较,实验组细胞转染PTTG-shRNA慢病毒载体后,PTTG mRNA和蛋白表达显著下降(均P<0.01),细胞生长显著变慢(均P<0.05),G0/G1期细胞比例显著增高(均P<0.05),凋亡率明显增加(均P<0.05)。结论慢病毒介导shRNA沉默PTTG基因表达可显著抑制人生长激素型垂体腺瘤细胞的生长,为进一步研究垂体腺瘤的发病机制和肿瘤的基因治疗提供实验基础。
Objective To observe the effects of shRNA-associated PTTG silencing mediated with lentiviral vectors on the growth of human growth hormone-secreting pituitary adenoma cells. Methods The human growth hormone-secreting pituitary adenoma cells were cultured primarily and divided into normal control group, negative control group and experimental group. The tumor cells were transfected by lentiviral vector of PTTG-shRNA consmJcted in experimental group and by blank lentiviral vector in negative control group. No transfection was executed in normal control group. The transfection efficiency of tumor cells was analyzed by flow cytometer （FCM）. Expressions of PTTG mRNA and protein were analyzed by RT-PCR and Western blot respectively. Cell proliferation was detected by MTT. The change of cell cycle was determined by FCM. Results Compared with the normal control group and negative control group, the expressions of PTTG mRNA and protein significantly decreased in experimental group after the transfection of PTTG-shRNA lentiviral vector （all P 〈 0.01）, the proliferation of pituitary adenoma cells was inhibited （both P 〈 0.05）, the cell proportion of GdGI phase markedly rose （both P 〈 0.05）, and the apoptosis rate obviously increased （both P 〈 0.05）. Conclusions shRNA-associated PTTG silencing mediated by lentiviral vector can significantly inhibit the growth of human growth hormone-secreting pituitary adenoma cells, which provides experimental basis for further research of pathogenesis and gene therapy for pituitary adenomas.
出处
《中国微侵袭神经外科杂志》
CAS
2012年第8期376-378,共3页
Chinese Journal of Minimally Invasive Neurosurgery
关键词
垂体肿瘤
短发夹RNA
癌基因
细胞分裂
pituitary neoplasms
short hairpin RNA
oncogenes
cell division
