摘要
目的探讨外源性野生型PTEN cDNA对人子宫内膜癌细胞RL95—2增殖、克隆形成和凋亡的影响。方法脂质体介导携带野生型VIEN cDNA的质粒pcDNA3．0-PTENcDNA转染人子宫内膜癌细胞RL95—2；G418筛选被转染的细胞株并扩增培养；RT—PCR方法检测PTEN cDNA在RL95—2细胞中的表达，并以转染空载体及未转染的RL95—2为对照组。四唑盐（MTT ）比色实验检测细胞增殖活力，软琼脂克隆形成实验检测转染后细胞的克隆形成率，透射电镜观察细胞形态。结果C-418筛选得到稳定表达细胞株；RT—PCR方法检测PTEN cDNA在RL95—2细胞中持续表达。转染PTEN cDNA组的细胞培养24h、48h、72h和96h后‰值明显低于转染空载体组（P〈0．01），而转染空载体组的A4帅值与未转染组比较无明显差异（P〉0．05）；转染PTEN cDNA组细胞克隆形成数与转染空载体组之间相比有显著性差异（P〈0．01），而转染空载体组与未转染组之间相比没有显著性差异（P〉0．05）；转染PTEN cDNA组透射电镜下出现肿瘤细胞凋亡表现，而转染空载体组细胞超微结构未见异常。结论PTEN cDNA稳定表达可明显抑制RL95—2细胞的体外增殖、克隆形成并诱导细胞凋亡。
Objective To study the effects of exogenous wild - type PIEN（phosphatase and tensin homolng deleted on chromosome ten） eDNA on in vitro proliferation colony-forming efficiency and apoptosis of the human endometrial carcinoma cells RL95- 2.Methods The plasmids including wild type PTEN cDNA, pcDNA3.0- PTEN eDNA were transferred with lipefectamine into in vitro cultured human endometrial carcinoma cells RL95- 2.After G418 selection,RL95 - 2 cells infected were obtained and amplified.The expression of target genes was detected by RT- PCR. Cell viability was determined by MTT assay. Colony - forming efficiency behaviors of RL95 - 2 cells were tested by soft agar cloning method. The ultrastrueture of the cells were observed under electronic microscope. Results The cells infected were succesefully obtained by G418 selection assay. The steady expression of PTEN cDNA in human RL95 - 2 cells were verified by RT- PCR. After culture for 24 h,48 h,72 h and 96 h,the number of A490 of the group transfected with PTEN eDNA were （0.1 225 ± 0.035 104）, （0.1 905 ± 0.033 822）,（0.3 040±0.022 839）,and （0.327 667 ±0.11 892）, respectively, which means that the group transfected with PTEN eDNA had a much lower cell viability than other ones （P 〈 0.01 ）. The colony forming number of the group transfected with PTEN cDNA was （ 11.8 ± 1.92）,which was significantly decreased than that of the control group （ P 〈 0.01）. Some classical morphological changes of apoptesis were observed in the group transfected with PTEN cDNA under electronic microscope and none in the control group. Conclusions The expression of PTEN cDNA suppresses the in vitro growth of human endometrial carcinoma cells, decrease the colony - forming efficiency and induces RL95 - 2 cells to apoptosis. PTEN gene may be a novel therapeutic agent for endometrial carcinoma.
出处
《武警医学》
CAS
2008年第10期894-897,F0003,共5页
Medical Journal of the Chinese People's Armed Police Force
关键词
PTEN
子宫内膜癌
增殖
凋亡
PTEN gene Endometrial carcinoma Proliferation Apoptosis
