survivin基因修饰后对卵巢上皮性癌HO-8910细胞周期及化疗敏感性的影响  

Influence of human epithelial ovarian cancer HO-8910 cells with modified survivin gene on the cell cycle distribution and chemosensitivity

作　　者：彭立平[1] 黄建鸣[2] 张国楠[1] 查晓[2] 任源[2] 樊英[1] 邓碧芳[2] 

机构地区：[1]四川省肿瘤医院妇瘤科,成都610041 [2]四川省肿瘤医院基础研究部,成都610041

出　　处：《中华妇产科杂志》2010年第11期860-864,共5页Chinese Journal of Obstetrics and Gynecology

摘　　要：目的 探讨survivin基因经不同方式基因修饰后对卵巢上皮性癌（卵巢癌）细胞系HO-8910细胞周期和化疗敏感性的影响.方法 对survivin基因进行两种方式的基因修饰,即survivin基因核苷酸序列第34位苏氨酸突变为丙氨酸（T34A）和N末端缺失8个氨基酸（N-8AA）,构建含survivin^N-8AA和survivin^T34A基因的表达载体,将含survivin^N-8AA、survivin^T34A和野生型survivin基因的表达载体转染HO-8910细胞（分别为SN-HO-8910组、M-HO-8910组和Sur-HO-8910组）,以转染空载体pcDNA3.1质粒（PC-HO-8910组）为对照.（1）采用逆转录（RT）PCR技术检测和测序法鉴定转染后HO-8910细胞各目的 基因mRNA的表达;（2）采用流式细胞仪检测各组转染后HO-8910细胞的细胞周期比例;（3）采用四甲基偶氮唑蓝（MTT）比色法检测各组转染后HO-8910细胞对顺铂、紫杉醇和磷脂酰肌醇3激酶（PI3K）抑制剂--LY294002的敏感性[以50%抑制浓度（IC50）表示].结果 （1）RT-PCR技术检测和测序法鉴定结果显示,转染后的HO-8910细胞分别稳定表达survivin^T34A、survivin^N-8AA和野生型survivin mRNA.（2）SN-HO-8910组细胞G0/G1期比例为44.72%,明显低于PC-HO-8910组的49.64%（P＜0.05）;而G2/M期和S期比例（分别为1.06%和54.22%）明显高于PC-HO-8910组（分别为0.56%和49.80%;P＜0.05）.M-HO-8910组细胞G2/M期和S期比例分别为0.16%和36.33%,明显低于PC-HO-8910组（P＜0.05）;而G0/G1期比例（63.51%）明显高于PC-HO-8910组（P＜0.05）.Sur-HO-8910组G0/G1期、G2/M期和S期比例分别为54.46%、0.62%、44.92%,分别与PC-HO-8910组比较,差异均无统计学意义（P＞0.05）.（3）Sur-HO-8910组细胞对顺铂和紫杉醇的IC50[分别为（20.4±6.1）和（36.7±4.0）μmol/L]均明显高于PC-HO-8910组[分别为（14.4±3.9）和（28.6±3.6）μmol/L;P＜0.05].SN-HO-8910组细胞对顺铂和LY294002的IC50[分别为（7.6±1.0）和（13.2±4.0）μmol/L]均明显低于PC-HO-8910组[分别为（14.4±Objective To study the influence of survivin mutant-T34A （ survivin^T34A） and survivin deletant-N-terminal 8 amino acids residues （ survivin^N-8AA ） on the cell cycle distribution and chemosensitivity in human ovarian cancer HO-8910 cells for explorating the roles of modified survivin-mediated apoptosis induced by chemotherapeutic agents and possible signaling pathways involved. Methods pcDNA3.1 plasmid contained wild-type, survivin^T34A and survivin^N-8AA genes were transfected into HO-8910 cells,respectively, the control groups were HO-8910 cells transfected with pcDNA3. 1 plasmids. The expression of mRNA was examined by reverse transcription（RT） PCR and identified by DNA sequencing; the cell cycles were determined by flow cytometer analysis （ FCM ）; the growth inhibitions rate of cisplatin （ DDP）,paclitaxel （PTX） and LY294002 on the transfected cells were determined using methyl thiazolyl tetrazolium （MTT） assay. Results （1） The RT-PCR procedures and genome sequences showed that the survivin mRNA were expressed stable in the transfected HO-8910 cells. （2） There was lower percent of G0/G1 phase cells in SN-HO-8910 cells than that in PC-HO-8910 cells （44. 72% vs. 49.64%, P 〈0. 05） ;while higher percentage of G2/M phase and S phase cells（ 1.06% and 54. 22% vs. 0. 56% and 49. 80%, P 〈 0. 05 ）.There was lower the G2/M phase and S phase cells in M-HO-8910 cells 0. 16% and 36. 33%, than that in PC-HO-8910 cells（ P 〈 0. 05 ）; while higher percentage of G0/G1 phase cells（63. 51% ,P 〈 0. 05 ）. G0/G1 ,G2/M and S phase cells in Sur-HO-8910 cells were 54. 46%, 0. 62% and 44. 92%, and there were not significantly difference （ P 〉 0. 05 ）, compared to those in PC-HO-8910 cells. （ 3 ） The inhibitory concentration （ IC50 ） of DDP and PTX were higher in Sur-HO-8910 cells than those in control cells [（20. 4 ±6. 1）vs. （14.4 ±3.9）μmol/L,（36.7 ±4.0） vs. （28.6 ±3.6） μmol/L;all P〈0.05]. The IC50 of DDP and LY294002 in SN-HO-8910 cells we

关 键 词：微管相关蛋白质类 卵巢肿瘤 细胞系 肿瘤 细胞周期 顺铂 色酮类 吗啉类 

分 类 号：R686[医药卫生—骨科学]