摘要
目的:研究livin特异性小分子干扰RNA (siRNA)沉默胃癌SGC-7901细livin基因及对促胃癌细胞凋亡的影响。方法:设计针对人源livin基因的两条SiRNA:si-livin1和si-livin2,分别转染至对数生长期的胃癌SGC-7901细胞:逆转录酶链式反应(RT- PCR)检测转染前后胃癌SGC-7901细胞livin基因表达的变化,MTT法检测转染前后该细胞对5-FU、顺铂的半数抑制浓度(IC50)的变化以及检测转染前后细胞增殖能力的变化,PI染色后流式细胞仪检测转染前后细胞凋亡的变化。结果:livin特异性siRNA转染胃癌SGC-7901细胞48h后,si-livin1组livinαmRNA(灰度值表示)表达较空白对照组和阴性对照组显著减少(livinα:0.167±0.013 vs 0.403±0.036,0.389±0.053;livinβ:0.181±0.028 vs 0.413±0.041,0.404±0.029,均尸<0.01),而si-livin2组livin mRNA表达相比于空白对照组和阴性对照无显著差异;转染siRNA后,si-livin1组胃癌细胞生长缓慢,生长曲线较对照组平缓;si-livin1组细胞对5-FU和顺铂半数抑制浓度IC50较空白对照组和阴性对照组显著降低(5-FU:34.07±2.98 vs 74.39±4.91,69.85±4.57;顺铂:4.56±0.35 vs 9.07±0.44,7.96±0.64,均P<0.01);转染siRNA后,si-livin1组胃癌细胞自发凋亡率较空白对照组和阴性对照组增加(11.07±1.36 vs 3.54±2.89,6.72±1.77,P<0.01,P<0.05).5-FU和顺铂作用后,si-livin1组胃癌细胞凋亡率较空白对照组和阴性对照组显著增加(5-FU:34.90±1.76 vs 11.54±O.83,13.54±2.55;顺铂:48.14±2.70 vs 14.51±0.35,15.71±0_34,均P<0.01).结论:RNA干扰可有效沉,livin基因表达从而抑制胃癌SGC-7901细胞生长及增加该细胞凋亡敏感性,livin可能成为胃癌凋亡治疗途径的一个分子靶点。
AIM： To explore the teasiDlltry o~ ~llta~ interfering RNA （siRNA） for inhibiting livin gene expression, and to investigate the apoptotic susceptibility of SGC-7901 cells to siRNA- mediated silencing of the livin gene.
METHODS： Two siRNAs （si-livinl and si-livin2） that specifically target the livin gene were designed based on their coding regions and were synthesized in vitro. They were then transfected into gastric cancer SGC-7901 cells. Expression level of livin mRNA was assayed by RT-PCR. Apoptosis was assessed by flow cytometry. Growth of SGC-7901 cells and 50% inhibitory concentration （IC50） of 5-fluorouracil （5-FU） and cisplatin for cell growth were analyzed by the MTT method.
RESULTS： The expression level of livin mRNA treated for 48 hours by si-livinl was significantly decreased in SGC7901 cells （livin αr, 0.167 ± 0.013 vs 0.403 ± 0.036, 0.389 ± 0.053; livin 9, 0.181 ± 0.028 vs 0.413 ± 0.041, 0.404 ± 0.029, all P 〈 0.01） with an inhibited cell growth state, but not in silivin2-treated cells. IC50 of 5-FU and cisplatin in SGC-7901 cells treated by siivinl was decreased （5-FU, 34.07 ± 2.98 vs 74.39 ± 4.91, 69.85± 4.57; cisplatin, 4.56 ± 0.35 vs 9.07 ± 0.44, 7.96± 0.64, all P 〈 0.01）. The spontaneous apoptosis rate of cells was increased when they were treated by si- Iivinl （11.07 ± 1.36 vs 3.54 ± 2.89, 6.72 ＋ 1.77, P 〈 0.01, P 〈 0.05）, and cells treated by siivinl were more susceptible to proapoptotic stimuli （5-FU and cisplatin） than control groups （5-FU, 34.90 ± 1.76 vs 11.54 ± 0.83, 13.54± 2.55; cisplatin, 48.14 ± 2.70 vs 14.51 ± 0.35, 15.71±0.34, all P〈0.01）.
CONCLUSION： siRNA can silence livin gene expression in SGC-7901 cells and induce cell apoptosis; thus, livin might serve as a new target for apoptosis-inducing therapy against gastric cancer.
出处
《世界华人消化杂志》
CAS
北大核心
2007年第22期2377-2381,共5页
World Chinese Journal of Digestology
基金
江苏省医学重点人才135工程资助项目
No.52-2001~~
