| 刘慧晴,李玄,孟星圻,王璇,赵宗惠,贺修胜,李素云.GRP78基因短发夹RNA表达载体的构建和鉴定.[J].中南医学科学杂志.,2021,(2):158-161. |
| GRP78基因短发夹RNA表达载体的构建和鉴定 |
| Construction and identification of short hairpin RNA expression vector of GRP78 gene |
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| DOI：10.15972/j.cnki.43-1509/r.2021.02.008 |
| 中文关键词: 胃癌 葡萄糖调节蛋白78 短发夹RNA 载体 [ |
| 英文关键词:gastric cancer GRP78 shRNA vctor |
| 基金项目: |
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| 中文摘要: |
| 目的构建葡萄糖调节蛋白78(GRP78)基因短发夹RNA表达载体并鉴定,为探讨GRP78对胃癌发生发展作用及机制提供前期基础。方法运用核糖核酸(RNA)干扰技术构建靶向干扰GRP78蛋白短发夹RNA(GRP78-shRNA)载体,载体经转化、抽提及测序,用脂质体将GRP78-shRNA载体瞬时转染至SGC-7901胃癌细胞中,应用荧光显微镜、反转录定量PCR(qRT-PCR)及免疫印迹(Western blot)观察GRP78表达情况。结果GRP78-shRNA载体测序显示序列正确；转染GRP78-shRNA载体的SGC-7901胃癌细胞可见绿色荧光；qRT-PCR和Western blot结果显示,转染GRP78-shRNA载体的SGC-7901胃癌细胞中GRP78基因和蛋白表达降低。结论成功构建并鉴定GRP78基因短发夹RNA表达载体。 |
| 英文摘要: |
| The short hairpin RNA expression vector of glucose regulatory protein 78 (GRP78) gene was constructed and identified, to provide a preliminary basis for exploring the effect and mechanism of GRP78 on the development of gastric cancer. MethodsThe GRP78 short hairpin Ribonucleic acid (GRP78-shRNA) vector was constructed by RNA interference technology. The vector was transformed, extracted and sequenced. The GRP78-shRNA vector was transiently transfected into SGC-7901 gastric cancer cell by liposome of lipofectamine 2000. The expression of GRP78 was observed by fluorescence microscopy, Quantitative Real-time PCR (qRT-PCR) and immunoblot(Western blot). ResultsThe sequence of GRP78-shRNA vector was correct. The green fluorescence was observed in SGC-7901 gastric cancer cell transfected with GRP78-shRNA vector. The qRT-PCR and Western blot results showed that the expression of GRP78 gene and protein was decreased in SGC-7901 gastric cancer cell transfected with GRP78-shRNA vector. ConclusionThe short hairpin RNA expression vector of GRP78 gene was successfully constructed and identified. |
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