一种嵌合型调控元件在肿瘤靶向基因治疗中的应用

doi:10.13560/j.cnki.biotech.bull.1985.2015.10.036

生物技术通报 ›› 2015, Vol. 31 ›› Issue (10): 255-262.doi: 10.13560/j.cnki.biotech.bull.1985.2015.10.036

一种嵌合型调控元件在肿瘤靶向基因治疗中的应用

周培杰¹,高维强^1,2,方煜翔¹

1.上海交通大学医学院附属仁济医院仁济-MedX临床干细胞研究中心癌基因及相关基因国家重点实验室, 上海 200127;
2.上海交通大学生物医学工程学院, 上海 200030

收稿日期:2015-03-11 出版日期:2015-10-28 发布日期:2015-10-28
作者简介:周培杰, 男, 硕士研究生, 研究方向：肿瘤学; E-mail：zhpejie@126.com
基金资助:
国家自然科学基金项目（81301857, 81130038）

An Integrated Gene Regulatory Element for Tumor-specific Targeting Therapy

Zhou Peijie¹, Gao Weiqiang^{1, 2}, Fang Yuxiang¹

1. State Key Laboratory of Oncogenes and Related Genes, Renji-MedX Clinical Stem Cell Research Center, Affiliated Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127; 2. School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai 200030

Received:2015-03-11 Published:2015-10-28 Online:2015-10-28

摘要/Abstract

摘要： 肿瘤靶向基因治疗成功的关键是调控治疗基因在肿瘤细胞中特异、高效地表达。首次构建一种嵌合型表达调控元件, 旨在转录水平、转录后水平和翻译水平上实现联合调控目的基因在肿瘤细胞中特异性表达。体外实验表明, 在前列腺癌细胞系LNCaP中, 该调控元件可将报告基因增强型绿色荧光蛋白（EGFP）和荧光素酶（luciferase）的肿瘤表达特异性分别提高420%和480%。体外细胞存活实验表明, 运用该元件调控单纯疱疹病毒-1胸腺激酶（HSV-1 TK）的表达能特异性杀伤LNCaP细胞, 验证了该元件可成功用于治疗基因的肿瘤靶向表达。

关键词: 肿瘤靶向基因治疗, 嵌合型表达调控条件, 绿色荧光蛋白, 单纯疱疹病毒-1胸腺激酶, 前列腺癌

Abstract: The key for a successful tumor-specific gene therapy is to regulate the therapeutic gene specifically and efficiently expressing in tumor cells. By the construction of an embedded expression regulatory element, a high efficiency as well as tumor-specific expression of target genes in tumor cells was achieved by integrated regulation at transcriptional, post-transcriptional and translational levels simultaneously. It was shown that the integrated gene regulatory element improved expressional specificities in prostate cancer cell line LNCaP up to 420% and 480% using enhanced green fluorescent protein（EGFP）and luciferase as reporter gene respectively in vitro. By survival assay in vitro, LNCaP cell survival was specifically suppressed by HSV-1 TK expression under the control of this regulator. These results confirmed that this element could be applied in tumor-specific targeting therapy.

Key words: tumor-specific targeting therapy, embedded expression regulatory element, EGFP, HSV-1 TK, prostate cancer

周培杰,高维强,方煜翔. 一种嵌合型调控元件在肿瘤靶向基因治疗中的应用[J]. 生物技术通报, 2015, 31(10): 255-262.

Zhou Peijie, Gao Weiqiang, Fang Yuxiang. An Integrated Gene Regulatory Element for Tumor-specific Targeting Therapy[J]. Biotechnology Bulletin, 2015, 31(10): 255-262.

参考文献

[1]Lie-A-Ling M, Bakker CT, Deurholt T, et al. Selection of tumour specific promoters for adenoviral gene therapy of cholangiocarcinoma[J]. J Hepatol, 2006, 44（1）：126-133.
[2]Foka P, Pourchet A, Hernandez AR, et al. Novel tumour-specific promoters for transcriptional targeting of hepatocellular carcinoma by herpes simplex virus vectors[J]. J Gene Med, 2010, 12（12）：956-967.
[3]Fang L, Shanqu L, Ping G, et al. Gene therapy with RNAi targeting UHRF1 driven by tumor-specific promoter inhibits tumor growth and enhances the sensitivity of chemotherapeutic drug in breast cancer in vitro and in vivo[J]. Cancer Chemother Pharmacol, 2012, 69（4）：1079-1087.
[4]Lukowski SW, Rothnagel JA, Trezise AE. CFTR mRNA expression is regulated by an upstream open reading frame and RNA secondary structure in its 5' untranslated region[J]. Hum Mol Genet, 2015, 24（4）：899-912.
[5]Wolfe AL, Singh K, Zhong Y, et al. RNA G-quadruplexes cause eIF4A-dependent oncogene translation in cancer[J]. Nature, 2014, 513（7516）：65-70.
[6]Spilka R, Ernst C, Mehta AK, et al. Eukaryotic translation initiation factors in cancer development and progression[J]. Cancer Lett, 2013, 340（1）：9-21.
[7]DeFatta RJ, Li Y, De Benedetti A. Selective killing of cancer cells based on translational control of a suicide gene[J]. Cancer Gene Ther, 2002, 9（7）：573-578.
[8]Moussavi M, Moshgabadi N, Fazli L, et al. Fibroblast growth factor and ornithine decarboxylase 5'UTRs enable preferential expression in human prostate cancer cells and in prostate tumors of PTEN（-/-）transgenic mice[J]. Cancer Gene Ther, 2012, 19（1）：19-29.
[9]Cipollini M, Landi S, Gemignani F. MicroRNA binding site polymorphisms as biomarkers in cancer management and research[J]. Pharmgenomics Pers Med, 2014, 7：173-1791.
[10]Valinezhad Orang A, Safaralizadeh R, Kazemzadeh-Bavili M. Mechanisms of miRNA-mediated gene regulation from common downregulation to mRNA-specific upregulation[J]. Int J Genomics, 2014：970607.
[11]Mazzacurati L, Marzulli M, Reinhart B, et al. Use of miRNA response sequences to block off-target replication and increase the safety of an unattenuated, glioblastoma-targeted oncolytic HSV[J]. Mol Ther, 2015, 23（1）：99-107.
[12]Li JM, Kao KC, Li LF, et al. MicroRNA-145 regulates oncolytic herpes simplex virus-1 for selective killing of human non-small cell lung cancer cells[J]. Virol J, 2013, 10：241.
[13]Lai YH, Lin CC, Chen SH, et al. Tumor-specific suicide gene therapy for hepatocellular carcinoma by transcriptionally targeted retroviral replicating vectors[J]. Gene Ther, 2015, 22（2）：155-162.
[14]Habibie, Yokoyama S, Abdelhamed S, et al. Survivin suppression through STAT3/β-catenin is essential for resveratrol-induced melanoma apoptosis[J]. Int J Oncol, 2014, 45（2）：895-901.
[15]Kogo R, How C, Chaudary N, et al. The microRNA-218~Survivin axis regulates migration, invasion, and lymph node metastasis in cervical cancer[J]. Oncotarget, 2015, 6（2）：1090-1100.
[16]Paik S, Shak S, Tang G, et al. A multigene assay to predict recurrence of tamoxifen-treated, node-negative breast cancer[J]. N Engl J Med, 2004, 351（27）：2817-2826.
[17]Hynes NE, Stoelzle T. Key signalling nodes in mammary gland development and cancer：Myc[J]. Breast Cancer Res, 2009, 11（5）：210.
[18]Meijer TW, Kaanders JH, Span PN, et al. Targeting hypoxia, HIF-1, and tumor glucose metabolism to improve radiotherapy efficacy[J]. Clin Cancer Res, 2012, 18（20）：5585-5594.
[19]Shi Y, Frost P, Hoang B, et al. MNK kinases facilitate c-myc IRES activity in rapamycin -treated multiple myeloma cells[J]. Oncogene, 2013, 32（2）：190-197.
[20]Lang KJ, Kappel A, Goodall GJ. Hypoxia-inducible factor-1alpha mRNA contains an internal ribosome entry site that allows efficient translation during normoxia and hypoxia[J]. Mol Biol Cell, 2002, 13（5）：1792-1801.
[21]Sujobert P, Bardet V, Cornillet-Lefebvre P, et al. Essential role for the p110delta isoform in phosphoinositide 3-kinase activation and cell proliferation in acute myeloid leukemia[J]. Blood, 2005, 106（3）：1063-1066.
[22]Fang Y, Xue JL, Shen Q, et al. MicroRNA-7 inhibits tumor growth and metastasis by targeting the phosphoinositide 3-kinase/Akt pathway in hepatocellular carcinoma[J]. Hepatology, 2012, 55（6）：1852-1862.

一种嵌合型调控元件在肿瘤靶向基因治疗中的应用

An Integrated Gene Regulatory Element for Tumor-specific Targeting Therapy

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