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一种表达Survivin-shRNA与融合自杀基因yCDglyTK的新型载体的构建及其在抑制结肠癌细胞增殖和迁移中的应用

Construction of a novel vector expressing Survivin-shRNA and fusion suicide gene yCDglyTK and its application in inhibiting proliferation and migration of colon cancer cells.

作者信息

Ye Ling, Yang Yuan, Ma Xin-Yu, Li Dan, Xu Mei-Li, Tan Pan, Long Li-Min, Wang Hai-Qin, Liu Ting, Guo Yong-Hong

机构信息

Department of Geriatrics, The Second Xiangya Hospital of Central South University, Changsha, Hunan 410011, P.R. China.

Department of Gastroenterology, Xiangya Hospital of Central South University, Changsha, Hunan 410008, P.R. China.

出版信息

Exp Ther Med. 2017 Nov;14(5):4721-4728. doi: 10.3892/etm.2017.5154. Epub 2017 Sep 20.

Abstract

Despite progress achieved in cancer chemotherapy in recent decades, adverse effects remain a limiting factor for a number of patients with colorectal cancer, suggesting the requirement for novel therapeutic strategies. Gene therapy appears to be a promising strategy for treating cancer. The present study aimed to investigate the anti-tumor effect of a combined gene therapy, using Survivin downregulation by RNAi and a fusion suicide gene yCDglyTK therapy system. A triple-gene vector expressing Survivin-targeted small hairpin RNA (Survivin-shRNA) and fusion suicide gene yCDglyTK was constructed, and administered to HCT116 cells. Survivin expression decreased significantly and yCDglyTK fusion gene expression was confirmed by both reverse transcription-quantitative polymerase chain reaction and western blot analysis. Introduction of Survivin-shRNA into yCDglyTK/prodrug system eradicated colon cancer cells and induced apoptosis more effectively. Furthermore, this therapeutic system is able to inhibit the migration of HCT116 cells. These results indicate that the recombinant plasmid may serve as a novel gene therapy approach to treat colorectal carcinoma.

摘要

尽管近几十年来癌症化疗取得了进展,但不良反应仍然是许多结直肠癌患者的限制因素,这表明需要新的治疗策略。基因治疗似乎是一种很有前景的癌症治疗策略。本研究旨在探讨一种联合基因治疗的抗肿瘤作用,该联合基因治疗采用RNA干扰下调Survivin以及融合自杀基因yCDglyTK治疗系统。构建了一种表达靶向Survivin的小发夹RNA(Survivin-shRNA)和融合自杀基因yCDglyTK的三基因载体,并将其应用于HCT116细胞。通过逆转录定量聚合酶链反应和蛋白质印迹分析证实,Survivin表达显著降低,yCDglyTK融合基因表达得到确认。将Survivin-shRNA引入yCDglyTK/前药系统能更有效地根除结肠癌细胞并诱导细胞凋亡。此外,该治疗系统能够抑制HCT116细胞的迁移。这些结果表明,重组质粒可作为一种治疗结直肠癌的新型基因治疗方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8018/5704315/5c3f12e0c6d4/etm-14-05-4721-g00.jpg

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