刘靖, 鲍依稀, 刘明学, 祝绚, 李进. RNA干扰enolase-1基因治疗胃癌的实验研究[J]. 中国肿瘤临床, 2008, 35(8): 455-458,462.
引用本文: 刘靖, 鲍依稀, 刘明学, 祝绚, 李进. RNA干扰enolase-1基因治疗胃癌的实验研究[J]. 中国肿瘤临床, 2008, 35(8): 455-458,462.
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LIU Jing, BAO Yixi, LIU Mingxue, ZHU Xuan, LI Jin. The Effect of RNA Interference of the Enolase-1 Gene on Gastric Carcinoma[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2008, 35(8): 455-458,462.
Citation: LIU Jing, BAO Yixi, LIU Mingxue, ZHU Xuan, LI Jin. The Effect of RNA Interference of the Enolase-1 Gene on Gastric Carcinoma[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2008, 35(8): 455-458,462.
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RNA干扰enolase-1基因治疗胃癌的实验研究

The Effect of RNA Interference of the Enolase-1 Gene on Gastric Carcinoma

    摘要
  • 摘要: 目的: 探讨RNA干扰enolase-1基因对人胃癌的治疗作用。 方法: 根据enolase-1mRNA序列构建表达enolase-1特异的小干扰RNA(small interfering RNA,siRNA)真核表达质粒pMSCVU6/ENO-1si1,pMSCVU6/ENO-1si2,pMSCVU6/ENO-1si3。种植人胃癌SGC-7901细胞于裸鼠皮下建立裸鼠肿瘤模型。将构建成功的三种siRNA质粒分别转染SGC-7901细胞株,pMSCVU6/ENO-1si2和pMSCVU6/ENO-1si3转染裸鼠瘤体内。MTT法检测SGC-7901细胞增殖状况;测量荷瘤裸鼠的肿瘤体积,计算抑瘤率;RT-PCR检测SGC-7901细胞enolase-1mRNA表达;分别以免疫组化SP法和Western blot检测体内外enolase-1蛋白表达。 结果: SGC-7901细胞株及裸鼠瘤体均被所采用的质粒成功转染。SGC-7901细胞受pMSCVU6/ENO-1si2转染后,其生长活性受到明显抑制(P<0.01),enolase-1mRNA及蛋白表达显著降低(P<0.01)。体内抑瘤实验表明,pMSCVU6/ENO-1si2组平均瘤体积为(380±26)mm3,pMSCVU6/ENO-1si3组与空白对照组分别为(993±124)mm3和(1102±131)mm3;pMSCVU6/ENO-1si2组抑瘤率为65.52%,pMSCVU6/ENO-1si3组为9.89%;pMSCVU6/ENO-1si2组肿瘤体积明显小于pMSCVU6/ENO-1si3组和空白对照组,差异具有显著性(P<0.01);pMSCVU6/ENO-1si3组和空白对照组之间肿瘤体积无显著差异(P>0.05);pM-SCVU6/ENO-1si2组与pMSCVU6/ENO-1si3组和空白对照组相比,enolase-1蛋白表达减弱。 结论: 表达siRNA的enolase-1基因特异的真核表达质粒能成功转染体内、外胃癌细胞,其中pMSCVU6/ENO-1si2可有效抑制人胃癌细胞的生长。

     

    Abstract: Objective: To assess the effect of RNA interference of enolase-1 mRNA in the treatment of gastric carcinoma. Methods: Three types of plasmids were synthesized: pMSCVU6/ENO-1si1, pMSCVU6/ENO-1si2, and pMSCVU6/ENO-1si3. The tumor model was established in nude mice with subcutaneous implantation of SGC-7901 gastric carcinoma cells.pMSCVU6/ENO-1si1, pMSCVU6/ENO-1si2 and pMSCVU6/ENO-1si3 were transfected into SGC-7901 cells. pMSCVU6/ENO-1si2 and pMSCVU6/ENO-1si3 were transfected into the transplanted tumors. The proliferation of SGC-7901 cells was detected by MTT assay. The growth inhibiting rate was calculated. Enolase-1 mRNA expression in SGC-7901 cells was detected by RT-PCR. Western blot and immunohistochemistry were employed to confirm enolase-1 protein expression in vitro and in vivo. Results: After the SGC-7901 cells were transfected with the plasmids, the viability of cells was in-hibited by pMSCVU6/ENO-1si2 (P<0.01). pMSCVU6/ENO-1si2 decreased the level of enolase-1 mRNA and protein ex-pression in SGC-7901 cells(P<0.01). In vivo experiments demonstrated that the tumor volume was 380± 26mm3 in the pM-SCVU6/ENO-1si2 group, 993± 124mm3 in the pMSCVU6/ENO-1si3 group, and 1102± 131mm3 in the control group. The growth inhibiting rate was 65.52% in the pMSCVU6/ENO-1si2 group and 9.89% in the pMSCVU6/ENO-1si3 group. The tumor volume in the pMSCVU6/ENO-1si2 group was smaller than that in the pMSCVU6/ENO-1si3 group and the control group(P<0.01). No significant difference was found in tumor volume between the pMSCVU6/ENO-1si3 group and the con-trol group (P>0.05). Enolase-1 protein expression was depressed in the pMSCVU6/ENO-1si2 group compared with that in the pMSCVU6/ENO-1si3group and the control group. pMSCVU6/ENO-1si2 significantly inhibited tumor growth in nude mice (P<0.01) and depressed enolase-1 protein expression in vivo. Conclusion: Small interfering RNA (siRNA) plasmids that express specific sequences of the enolase-1 gene can be successfully transfected into SGC-7901 cells in vivo and in vitro. Gastric carcinoma can be effectively inhibited by the plasmid pMSCVU6/ENO-1si2.

     

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