摘要
目的利用RNA干扰技术，研究靶向bcl-2的小干扰RNA（siRNA）在体内外对胃癌细胞生长的影响，探讨其对胃癌治疗的可行性。方法化学合成bcl-2基因的siRNA，脂质体法将bcl-2 siRNA转染胃癌SGC7901细胞，采用实时定量PCR和Western印迹观察bcl-2 siRNA转染前后胃癌细胞bcl-2基因的表达变化，四甲基偶氮唑盐（MTT）实验、流式细胞检测技术和端粒重复序列扩增法（TRAP）分别检测胃癌细胞增殖、凋亡和端粒酶活性变化。并将转染bcl-2 siRNA的胃癌SGC7901细胞接种于裸鼠皮下，观察其在裸鼠体内成瘤及生长情况。结果数据经方差分析，bcl-2 siRNA转染胃癌SGC7901细胞后，明显抑制bcl-2基因表达，并与其浓度和作用时间相关，以100nmol／L bcl-2 siRNA对bcl-2基因表达抑制效果最佳。以该浓度的bcl-2 siRNA转染胃癌SGC7901细胞后，bcl-2 mRNA和蛋白表达抑制分别为79．9％和85．3％；经bcl-2 siRNA作用的SGC7901细胞生长明显缓于对照组（P〈0．05），细胞凋亡率高于对照组（P〈0．05），端粒酶活性明显低于对照组（P〈0．05）。转染100nmol／L bcl-2 siRNA的胃癌SGC7901细胞接种裸鼠皮下后，肿瘤出现时间晚，体积小，生长受到明显抑制（P〈0．05）。结论靶向bcl-2的siRNA可明显下调靶基因bcl-2的表达，在体内外抑制胃癌SGC7901细胞的生长，为探索胃癌基因治疗提供新的策略。
Objective To determine the inhibitory effect of synthetic bcl-2 short interfering RNA （siRNA）on the expression of bcl-2 gene in human gastric cancer line SGC-7901 cells, and its effect on proliferation of SGC-7901 cells in vitro and in vivo, in order to probe the feasibility of gene therapy in this domain. Methods siRNA targeting bcl-2 mRNA was transfected into SGC-7901 cells, and bcl-2 expression was determined by real-time PCR and Western blot. Cell proliferation, apoptosis, and telomerase activity were examined by MTT and flow cytometry, and telomeric repeat amplification protocol assay, respectively. Gastric cancer cells treated by 100 nmol/L bcl-2 siRNA were subcutaneously transplanted into 5-week-old male nude mice and tumor growth was assessed. Results Bcl-2 siRNA significantly inhibited the expression of bcl-2 in gastric cancer cells at both mRNA and protein levels （79.9% and 85. 3%, respectively）, and the inhibition effect was characterized by time- and dosedependent transfection, and the concentration of 100 nmol/L for siRNA was regarded as the most effective. Telomerase activity of SGC-7901 cells treated with bcl-2 siRNA was lower than that of control cells （P〈0.05）, and the rate of apoptosis of SOC-7901 cells was higher than that of control groups （P〈 0.05）. The proliferation of SGC-7901 cells was also significantly suppressed both in vitro and in vivo （P〈0.05）. Conclusions siRNA directed against bcl-2 can significantly suppress proliferation of SGC- 7901 cells both in vitro and in vivo. This may provide a new gene therapy approach for gastric cancer.
出处
《中华消化杂志》
CAS
CSCD
北大核心
2007年第1期2-6,共5页
Chinese Journal of Digestion
关键词
胃肿瘤
RNA
小干扰
基因
BCL-2
Stomach neoplasms
RNA, small interferencing
Genes, bcl-2
