| 周洲朱金水△ 许志朋.慢病毒siRNA靶向干扰YAP基因胃癌细胞株的建立[J].,2011,11(4):605-610 |
| 慢病毒siRNA靶向干扰YAP基因胃癌细胞株的建立 |
| Establishment of Gastric Cancer Cells with Stable Interference bysiRNA Targeting YAP Gene |
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| DOI： |
| 中文关键词: YAP基因 慢病毒载体 胃癌细胞株SGC7901 RNA干扰 |
| 英文关键词: YAP gene Lentivirus vector Gastric cancer cell line SGC7901 RNA interference |
| 基金项目:上海市科委动物实验基金重点项目（10140902500） |
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| 中文摘要: |
| 目的：构建并鉴定YAP 基因短发夹干扰RNA(shRNA)慢病毒载体，建立稳定干扰YAP 基因表达的胃癌细胞株SGC7901。
方法：荧光定量PCR 检测YAP 基因在多种胃癌细胞株中的表达情况。构建重组靶向YAP 基因的shRNA 慢病毒表达质粒
PGC-shRNA-YAP，用脂质体转染的方法将载体导入胃癌细胞。经杀稻瘟菌素筛选后，建立稳定表达siRNA 的细胞株。荧光定量
PCR检测干扰效率。结果：在胃癌细胞株SGC7901中，YAP基因显示高表达。测序验证PGC-shRNA-YAP重组质粒构建成功。将
重组质粒稳定转染入胃癌细胞株SGC7901后能明显抑制YAPmRNA表达水平。结论：成功构建了PGC-shRNA-YAP慢病毒重组
质粒，建立了靶向稳定干扰YAP基因表达的siRNA胃癌细胞株SGC7901。 |
| 英文摘要: |
| Objective: To construct siRNA expression vector targeting YAP gene and establish the gastric cancer cells SGC7901
with stable inhibition of YAP gene. Methods: Rea-time PCR was used to examine the YAP gene expression in sevel gastric cancer cell
lines. The recombinant lentivirus PGC-shRNA-YAP vector was constructed, which was transfected into gastric cancer cells by Lipofectamine
2000. After gastric cancer cells with constant expression of the PGC-shRNA-YAP were established by blasticidin selection, the
interfere efficicncy of YAP mRNA expression level was detected by real-time PCR. Results: There was high expression of YAP gene in
the gastric cancer cell line SGC-7901. Recombinant lentivirus PGC-shRNA-YAP vector was successfully constructed by the identification
of sequencing. The recombinant vectormarkedly inhibited the expression ofYAP gene in gastric cancer cell line SGC-7901. Conclusion:
Recombinant lentivirus vector PGC-shRNA-YAP was successfully constructed and gastric cancer cell line SGC7901 with stable siRNA
expression targeting YAP gene was established. |
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