PTEN cDNA抑制子宫内膜癌细胞增殖作用的研究  

Effects of PTEN cDNA on cell proliferation of endometrial carcinoma cell line

作　　者：杨培丽[1] 杨选平[1] 刘玉环[2] 崔英[2] 

机构地区：[1]武警北京总队医院妇产科,北京100027 [2]第二军医大学长海医院妇产科,上海200433

出　　处：《武警医学院学报》2009年第3期208-212,F0003,共6页Acta Academiae Medicinae CPAPF

摘　　要：【目的】研究外源性PTEN cDNA对子宫内膜癌细胞生长增殖的影响,探讨子宫内膜癌治疗的新途径。【方法】经携带野生型PTEN cDNA的重组腺病毒(Ad-PTEN)介导将PTEN cDNA转入人子宫内膜癌细胞RL95-2,用反向聚合酶链反应(RT-PCR)检测出PTEN cDNA在RL95-2细胞中持续表达,X-gal染色方法测定腺病毒转染效率,台盼蓝染色活细胞计数绘制生长曲线,流式细胞分析仪(FCM)测定细胞周期。受试细胞随机分成(1)Ad-PTEN组:转染复制缺陷型腺病毒Ad-PTEN。(2)Ad-CMV组:转染空载体腺病毒Ad-CMV。(3)对照组:仅用培养基而不加腺病毒。【结果】Ad-PTEN感染RL95-2细胞后,RT-PCR方法检测PTEN cDNA在RL95-2细胞中持续表达;X-gal染色方法测得Ad-PTEN最佳感染复数为100。经台盼蓝染色活细胞计数后绘制细胞生长曲线,见经Ad-PTEN感染的细胞生长速度明显低于空载体病毒Ad-CMV感染及未经病毒感染的RL95-2细胞(P<0.01),而空载体Ad-CMV感染后的细胞与未经感染的RL95-2细胞比较,生长速度无差异(P>0.05);细胞周期分析表明,感染Ad-PTEN后的RL95-2细胞G1期细胞比例明显增多,而S期G2-M期的细胞减少(P<0.05)。【结论】Ad-PTEN介导的PTEN cDNA在子宫内膜癌细胞RL95-2中有表达。PTEN cDNA在RL95-2的中表达能够抑制细胞的增殖,将细胞周期阻断在G1期。[Objective] To study the effects of the exogenous wild-type PTEN （Phosphatase and Tensin homolog deleted on chromoseme Ten） cDNA on the prohfiration of human endometrial carcinoma cells RL95 - 2. [Methods] The wild type PTEN cDNA were transferred by recombinant adenovirus containing FFEN eDNA （Ad-PTEN） into cultured human endometrial earcinoma cells RL95 - 2. The steady expression of PTEN eDNA was detected by RT-PCR assy and the efficiency of Ad-PTEN trartsfecdon was deteoted by the method of X-gal stain. CeLl growth curve was obtained by viable count after trypan bluc stain. Cell cycle was determined by flow cytometry （FCM） . The cells for experiment were divided into 3 group by random. （1） experimental group which was infected by Ad-CMV. （2） adenovirus control group which was infected by Ad-CMV and （3） blank control group which was fock infected by medium. [Results] After transfection by Ad-PTEN the steady expression of PTEN cDNA in RL95-2 cells were vetificatianed by RT-PCR. The number of multiplicity of infection was 100. In comparison with the control group, the cell growth curve proved that growth velocity of the experimental group was low than the control groups, the differences were statistically significant （ P 〈 0.01 ） . Cell cycle analysis showed that cell was retarded and many cells were arrested in G1 phasc, the differences were statistically significant （P 〈 0.05） . [Conclusions] PTEN eDNA can be steady expressed in RL95-2 cells after transfection by Ad-PTEN. The expression of PTEN cDNA can suppress the growth of human endometrial carcinoma cells, and arrest the cell cycle in G1 phase. PTEN gene may be a novel therapeutic agent for endometrial carcinoma.

关 键 词：子宫内膜癌 PTEN 细胞增殖 细胞周期 

分 类 号：Q786[生物学—分子生物学]