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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.

DOI:10.3892/etm.2017.5154
PMID:29201172
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5704315/
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/cfc72c3c7358/etm-14-05-4721-g04.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8018/5704315/5c3f12e0c6d4/etm-14-05-4721-g00.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8018/5704315/d85f6e0f3b8e/etm-14-05-4721-g01.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8018/5704315/15bebb3a339e/etm-14-05-4721-g02.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8018/5704315/5ae099822a8b/etm-14-05-4721-g03.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8018/5704315/cfc72c3c7358/etm-14-05-4721-g04.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8018/5704315/5c3f12e0c6d4/etm-14-05-4721-g00.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8018/5704315/d85f6e0f3b8e/etm-14-05-4721-g01.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8018/5704315/15bebb3a339e/etm-14-05-4721-g02.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8018/5704315/5ae099822a8b/etm-14-05-4721-g03.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8018/5704315/cfc72c3c7358/etm-14-05-4721-g04.jpg

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本文引用的文献

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Influence of survivin-targeted therapy on chemosensitivity in the treatment of acute myeloid leukemia.生存素靶向治疗对急性髓系白血病治疗中化疗敏感性的影响。
Cancer Lett. 2015 Oct 1;366(2):160-72. doi: 10.1016/j.canlet.2015.05.033. Epub 2015 Jun 26.
2
Survivin: A molecular biomarker in cancer.生存素:癌症中的一种分子生物标志物。
Indian J Med Res. 2015 Apr;141(4):389-97. doi: 10.4103/0971-5916.159250.
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ARE/SUZ12 dual specifically-regulated adenoviral TK/GCV system for CML blast crisis cells.用于慢性粒细胞白血病急变期细胞的ARE/SUZ12双重特异性调控腺病毒TK/GCV系统
J Exp Clin Cancer Res. 2015 May 28;34(1):56. doi: 10.1186/s13046-015-0139-4.
4
Progress and problems with the use of suicide genes for targeted cancer therapy.用于靶向癌症治疗的自杀基因的进展与问题
Adv Drug Deliv Rev. 2016 Apr 1;99(Pt A):113-128. doi: 10.1016/j.addr.2015.05.009. Epub 2015 May 22.
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Downregulation of survivin inhibits proliferation and migration of human gastric carcinoma cells.生存素的下调抑制人胃癌细胞的增殖和迁移。
Int J Clin Exp Pathol. 2015 Feb 1;8(2):1731-6. eCollection 2015.
6
Evaluation of the toxicity of anticancer chemotherapy in patients with colon cancer.结肠癌患者抗癌化疗毒性评估
Adv Clin Exp Med. 2015 Jan-Feb;24(1):103-11. doi: 10.17219/acem/38154.
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Survivin over-expression is correlated with a poor prognosis in esophageal cancer patients.生存素过表达与食管癌患者的不良预后相关。
Clin Chim Acta. 2015 Jun 15;446:82-5. doi: 10.1016/j.cca.2015.04.009. Epub 2015 Apr 17.
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Inducible RNAi system and its application in novel therapeutics.诱导性 RNAi 系统及其在新型治疗中的应用。
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Survivin - The inconvenient IAP.存活素——颇具争议的凋亡抑制蛋白
Semin Cell Dev Biol. 2015 Mar;39:91-6. doi: 10.1016/j.semcdb.2014.12.007. Epub 2015 Jan 12.
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J Control Release. 2015 Feb 28;200:179-87. doi: 10.1016/j.jconrel.2015.01.003. Epub 2015 Jan 7.