Full Text:   <713>

Summary:  <248>

Suppl. Mater.: 

CLC number: 

On-line Access: 2022-11-15

Received: 2022-03-08

Revision Accepted: 2022-07-22

Crosschecked: 2022-11-16

Cited: 0

Clicked: 1065

Citations:  Bibtex RefMan EndNote GB/T7714




Yong HAN


Lin XU


-   Go to

Article info.
Open peer comments

Journal of Zhejiang University SCIENCE B 2022 Vol.23 No.11 P.915-930


Optimized thyroid transcription factor-1 core promoter-driven microRNA-7 expression effectively inhibits the growth of human non-small-cell lung cancer cells

Author(s):  Shipeng CHEN, Lian GUAN, Xu ZHAO, Jing YANG, Longqing CHEN, Mengmeng GUO, Juanjuan ZHAO, Chao CHEN, Ya ZHOU, Yong HAN, Lin XU

Affiliation(s):  Special Key Laboratory of Gene Detection and Therapy & Base for Talents in Biotherapy of Guizhou Province, Zunyi 563000, China; more

Corresponding email(s):   xulinzhouya@163.com, Chinahotfog@126.com, zhouyazmc@163.com

Key Words:  Lung cancer, Thyroid transcription factor-1 (TTF-1), Promoter, MicroRNA-7 (miR-7), Nuclear factor-1 (NF-1)

Shipeng CHEN, Lian GUAN, Xu ZHAO, Jing YANG, Longqing CHEN, Mengmeng GUO, Juanjuan ZHAO, Chao CHEN, Ya ZHOU, Yong HAN, Lin XU. Optimized thyroid transcription factor-1 core promoter-driven microRNA-7 expression effectively inhibits the growth of human non-small-cell lung cancer cells[J]. Journal of Zhejiang University Science B, 2022, 23(11): 915-930.

@article{title="Optimized thyroid transcription factor-1 core promoter-driven microRNA-7 expression effectively inhibits the growth of human non-small-cell lung cancer cells",
author="Shipeng CHEN, Lian GUAN, Xu ZHAO, Jing YANG, Longqing CHEN, Mengmeng GUO, Juanjuan ZHAO, Chao CHEN, Ya ZHOU, Yong HAN, Lin XU",
journal="Journal of Zhejiang University Science B",
publisher="Zhejiang University Press & Springer",

%0 Journal Article
%T Optimized thyroid transcription factor-1 core promoter-driven microRNA-7 expression effectively inhibits the growth of human non-small-cell lung cancer cells
%A Shipeng CHEN
%A Lian GUAN
%A Jing YANG
%A Longqing CHEN
%A Mengmeng GUO
%A Juanjuan ZHAO
%A Chao CHEN
%A Yong HAN
%A Lin XU
%J Journal of Zhejiang University SCIENCE B
%V 23
%N 11
%P 915-930
%@ 1673-1581
%D 2022
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B2200116

T1 - Optimized thyroid transcription factor-1 core promoter-driven microRNA-7 expression effectively inhibits the growth of human non-small-cell lung cancer cells
A1 - Shipeng CHEN
A1 - Lian GUAN
A1 - Xu ZHAO
A1 - Jing YANG
A1 - Longqing CHEN
A1 - Mengmeng GUO
A1 - Juanjuan ZHAO
A1 - Chao CHEN
A1 - Ya ZHOU
A1 - Yong HAN
A1 - Lin XU
J0 - Journal of Zhejiang University Science B
VL - 23
IS - 11
SP - 915
EP - 930
%@ 1673-1581
Y1 - 2022
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B2200116

Targeted gene therapy has become a promising approach for lung cancer treatment. In our previous work, we reported that the targeted expression of microRNA-7 (miR-7) operated by thyroid transcription factor-1 (TTF-1) promoter inhibited the growth of human lung cancer cells in vitro and in vivo; however, the intervention efficiency needed to be further improved. In this study, we identified the core promoter of TTF-1 (from -1299 bp to -871 bp) by 5' deletion assay and screened out the putative transcription factors nuclear factor-1 (NF-1) and activator protein-1 (AP-1). Further analysis revealed that the expression level of NF-1, but not AP-1, was positively connected with the activation of TTF-1 core promoter in human non-small-cell lung cancer (NSCLC) cells. Moreover, the silencing of NF-1 could reduce the expression level of miR-7 operated by TTF-1 core promoter. Of note, we optimized four distinct sequences to form additional NF-1-binding sites (TGGCA) in the sequence of TTF-1 core promoter (termed as optTTF-1 promoter), and verified the binding efficiency of NF-1 on the optTTF-1 promoter by electrophoretic mobility shift assay (EMSA). As expected, the optTTF-1 promoter could more effectively drive miR-7 expression and inhibit the growth of human NSCLC cells in vitro, accompanied by a reduced transduction of NADH dehydrogenase (ubiquinone) 1α subcomplex 4 (NDUFA4)/protein kinase B (Akt) pathway. Consistently, optTTF-1 promoter-driven miR-7 expression could also effectively abrogate the growth and metastasis of tumor cells in a murine xenograft model of human NSCLC. Finally, no significant changes were detected in the biological indicators or the histology of some important tissues and organs, including heart, liver, and spleen. On the whole, our study revealed that the optimized TTF-1 promoter could more effectively operate miR-7 to influence the growth of human NSCLC cells, providing a new basis for the development of microRNA-based targeting gene therapy against clinical lung cancer.


概要:靶向基因治疗是一种很有前景的肺癌治疗方法。在前期工作中,我们报道了由甲状腺转录因子-1(TTF-1)启动子调控微小RNA-7(miR-7)的靶向表达可在体外和体内抑制人肺癌细胞的生长,但干预效率有待进一步提高。在本研究中,我们通过5’缺失分析鉴定了TTF-1的核心启动子(从−1299 bp到−871 bp),并筛选出潜在的结合转录因子核因子-1(NF-1)和激活蛋白-1(AP-1)。进一步分析的结果表明:NF-1的表达水平,而非AP-1的表达水平,与人非小细胞肺癌NSCLC细胞中TTF-1核心启动子的活性呈正相关。此外,NF-1的沉默可以降低由TTF-1核心启动子调控的miR-7的表达。重要的是,我们在TTF-1核心启动子的序列上优化了四个不同的序列(称为optTTF-1启动子)以形成额外的NF-1结合位点(TGGCA),并通过电泳迁移率实验(EMSA)分析验证了NF-1对optTTF-1启动子的结合效率。通进一步研究,结果显示optTTF-1启动子可更有效地驱动miR-7表达并在体外抑制人NSCLC细胞的生长,同时减少Ⅰ型辅酶脱氢酶1α亚复合物4(NDUFA4)/蛋白激酶B(Akt)通路转导。optTTF-1启动子驱动的miR-7表达同样也可有效地抑制人NSCLC异种移植模型中肿瘤细胞的体内生长和转移。最后,模型动物的心脏、肝脏和脾脏等重要组织和器官的生物学指标和组织学未见明显变化。总之,本研究揭示了优化的TTF-1启动子可更有效地调控miR-7表达来抑制人NSCLC细胞的生长,为开发基于miRNA的靶向基因治疗临床肺癌提供了新的实验基础。


Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


[1]AmreddyN, BabuA, MuralidharanR, et al., 2017. Polymeric nanoparticle-mediated gene delivery for lung cancer treatment. Top Curr Chem (Cham), 375(2):35.

[2]BillietC, PeetersS, DecaluwéH, et al., 2016. Postoperative radiotherapy for lung cancer: is it worth the controversy? Cancer Treat Rev, 51:10-18.

[3]BucherK, Rodríguez-BocanegraE, DauletbekovD, et al., 2021. Immune responses to retinal gene therapy using adeno-associated viral vectors-implications for treatment success and safety. Prog Retin Eye Res, 83:100915.

[4]CaoQ, MaoZD, ShiYJ, et al., 2016. MicroRNA-7 inhibits cell proliferation, migration and invasion in human non-small cell lung cancer cells by targeting FAK through ERK/MAPK signaling pathway. Oncotarget, 7(47):77468-77481.

[5]ChanBA, CowardJIG, 2013. Chemotherapy advances in small-cell lung cancer. J Thorac Dis, 5(S5):S565-S578.

[6]ChandlerRJ, VenturoniLE, LiaoJ, et al., 2021. Promoterless, nuclease-free genome editing confers a growth advantage for corrected hepatocytes in mice with methylmalonic acidemia. Hepatology, 73(6):2223-2237.

[7]ChenHZ, GuoMM, YueDX, et al., 2021. MicroRNA-7 negatively regulates Toll-like receptor 4 signaling pathway through FAM177A. Immunology, 162(1):44-57.

[8]ChenSP, WangY, LiDM, et al., 2022. Mechanisms controlling microRNA expression in tumor. Cells, 11(18):2852.

[9]DaninoYM, EvenD, IdesesD, et al., 2015. The core promoter: at the heart of gene expression. Biochim Biophys Acta, 1849(8):1116-1131.

[10]DuX, XiaoJJ, FuXF, et al., 2021. A proteomic analysis of Bcl-2 regulation of cell cycle arrest: insight into the mechanisms. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 22(10):839-855.

[11]DuX, ZhangJQ, LiuL, et al., 2022. A novel anticancer prop

[12]erty of Lycium barbarum polysaccharide in triggering ferroptosis of breast cancer cells. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 23(4):286-299.

[13]DziadziuszkoR, KrebsMG, de BraudF, et al., 2021. Updated integrated analysis of the efficacy and safety of entrectinib in locally advanced or metastatic ROS1 fusion-positive non-small-cell lung cancer. J Clin Oncol, 39(11):1253-1263.

[14]EvenDY, KedmiA, IdesesD, et al., 2017. Functional screening of core promoter activity. In: Gould D (Ed.), Mammalian Synthetic Promoters. Humana, New York, p.77-91.

[15]FukazawaT, MaedaY, DurbinML, et al., 2007. Pulmonary adenocarcinoma-targeted gene therapy by a cancer- and tissue-specific promoter system. Mol Cancer Ther, 6(1):244-252.

[16]FulzeleSV, ChatterjeeA, ShaikMS, et al., 2006. Inhalation delivery and anti-tumor activity of celecoxib in human orthotopic non-small cell lung cancer xenograft model. Pharm Res, 23(9):2094-2106.

[17]GuanL, ZhaoX, TangL, et al., 2021. Thyroid transcription factor-1: structure, expression, function and its relationship with disease. Biomed Res Int, 2021:9957209.

[18]HuangQM, ZengYM, LinHH, et al., 2017. Transfection with Livin and Survivin shRNA inhibits the growth and proliferation of non-small cell lung cancer cells. Mol Med Rep, 16(5):7086-7091.

[19]IkedaK, Shaw-WhiteJR, WertSE, et al., 1996. Hepatocyte nuclear factor 3 activates transcription of thyroid transcription factor 1 in respiratory epithelial cells. Mol Cell Biol, 16(7):3626-3636.

[20]KimJH, KimHS, KimBJ, et al., 2018. Prognostic impact of TTF-1 expression in non-squamous non-small-cell lung cancer: a meta-analysis. J Cancer, 9(22):4279-4286.

[21]KizzireK, KharghariaS, RiceKG, 2013. High-affinity PEGylated polyacridine peptide polyplexes mediate potent in vivo gene expression. Gene Ther, 20(4):407-416.

[22]Lara-GuerraH, RothJA, 2016. Gene therapy for lung cancer. Crit Rev Oncog, 21(1-2):115-124.

[23]LeeAY, ChoMH, KimS, 2019. Recent advances in aerosol gene delivery systems using non-viral vectors for lung cancer therapy. Expert Opin Drug Deliv, 16(7):757-772.

[24]LeiLY, ChenC, ZhaoJJ, et al., 2017. Targeted expression of miR-7 operated by TTF-1 promoter inhibited the growth of human lung cancer through the NDUFA4 pathway. Mol Ther Nucleic Acids, 6:183-197.

[25]LiC, WuX, ZhangW, et al., 2014. AEG-1 promotes metastasis through downstream AKR1C2 and NF1 in liver cancer. Oncol Res, 22(4):203-211.

[26]LiC, BrantE, BudakH, et al., 2021. CRISPR/Cas: a Nobel Prize award-winning precise genome editing technology for gene therapy and crop improvement. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 22(4):253-284.

[27]LiJR, ZhengYJ, SunGY, et al., 2014. Restoration of miR-7 expression suppresses the growth of Lewis lung cancer cells by modulating epidermal growth factor receptor signaling. Oncol Rep, 32(6):2511-2516.

[28]LiY, EggermontK, VanslembrouckV, et al., 2013. NKX2-1 activation by SMAD2 signaling after definitive endoderm differentiation in human embryonic stem cell. Stem Cells Dev, 22(9):1433-1442.

[29]Linnerth-PetrikNM, SantryLA, YuDL, et al., 2012. Adeno-associated virus vector mediated expression of an oncogenic retroviral envelope protein induces lung adenocarcinomas in immunocompetent mice. PLoS ONE, 7(12):e51400.

[30]LiuZL, JiangZM, HuangJY, et al., 2014. miR-7 inhibits glioblastoma growth by simultaneously interfering with the PI3K/ATK and Raf/MEK/ERK pathways. Int J Oncol, 44(5):1571-1580.

[31]LuxCT, ScharenbergAM, 2017. Therapeutic gene editing safety and specificity. Hematol Oncol Clin North Am, 31(5):787-795.

[32]MassaroG, HughesMP, WhalerSM, et al., 2020. Systemic AAV9 gene therapy using the synapsin I promoter rescues a mouse model of neuronopathic Gaucher disease but with limited cross-correction potential to astrocytes. Hum Mol Genet, 29(12):1933-1949.

[33]NakazatoM, ChungHK, UlianichL, et al., 2000. Thyroglobulin repression of thyroid transcription factor 1 (TTF-1) gene expression is mediated by decreased DNA binding of nuclear factor I proteins which control constitutive TTF-1 expression. Mol Cell Biol, 20(22):8499-8512.

[34]NaldiniL, 2015. Gene therapy returns to centre stage. Nature, 526(7573):351-360.

[35]NasimF, SabathBF, EapenGA, 2019. Lung cancer. Med Clin North Am, 103(3):463-473.

[36]PhilpottC, TovellH, FraylingIM, et al., 2017. The NF1 somatic mutational landscape in sporadic human cancers. Hum Genomics, 11:13.

[37]PowellSK, Rivera-SotoR, GraySJ, 2015. Viral expression cassette elements to enhance transgene target specificity and expression in gene therapy. Discov Med, 19(102):49-57.

[38]PrullerJ, HoferI, GanassiM, et al., 2021. A human Myogenin promoter modified to be highly active in alveolar rhabdomyosarcoma drives an effective suicide gene therapy. Cancer Gene Ther, 28(5):427-441.

[39]QianB, WangDM, GuXS, et al., 2018. LncRNA H19 serves as a ceRNA and participates in non-small cell lung cancer development by regulating microRNA-107. Eur Rev Med Pharmacol Sci, 22(18):5946-5953.

[40]RaiKM, TakigawaN, ItoS, et al., 2011. Liposomal delivery of microRNA-7-expressing plasmid overcomes epidermal growth factor receptor tyrosine kinase inhibitor-resistance in lung cancer cells. Mol Cancer Ther, 10(9):1720-1727.

[41]SebestyénMG, BudkerVG, BudkerT, et al., 2006. Mechanism of plasmid delivery by hydrodynamic tail vein injection. I. Hepatocyte uptake of various molecules. J Gene Med, 8(7):852-873.

[42]SheikhS, ErnstD, KeatingA, 2021. Prodrugs and prodrug-activated systems in gene therapy. Mol Ther, 29(5):1716-1728.

[43]SherYP, TzengTF, KanSF, et al., 2009. Cancer targeted gene therapy of BikDD inhibits orthotopic lung cancer growth and improves long-term survival. Oncogene, 28(37):3286-3295.

[44]SuJC, RuanSL, DaiSK, et al., 2019. NF1 regulates apoptosis in ovarian cancer cells by targeting MCL1 via miR-142-5p. Pharmacogenomics, 20(3):155-165.

[45]TolozaEM, MorseMA, LyerlyHK, 2006. Gene therapy for lung cancer. J Cell Biochem, 99(1):1-22.

[46]UmakanthanS, BukeloMM, 2021. Concise genetic profile of lung carcinoma. Postgrad Med J, 0:1-5.

[47]UmakanthanS, RaoAVC, MohammedW, 2021. Role of immunohistochemistry markers in neoplastic lung lesions. J Cancer Res Ther, 17(6):1382-1388.

[48]WigginsJF, RuffinoL, KelnarK, et al., 2010. Development of a lung cancer therapeutic based on the tumor suppressor microRNA-34. Cancer Res, 70(14):5923-5930.

[49]WilsonC, NimickM, NehoffH, et al., 2017. ALK and IGF-1R as independent targets in crizotinib resistant lung cancer. Sci Rep, 7:13955.

[50]XieXM, HsuJL, ChoiMG, et al., 2009. A novel hTERT promoter-driven E1A therapeutic for ovarian cancer. Mol Cancer Ther, 8(8):2375-2382.

[51]XiongSD, ZhengYJ, JiangP, et al., 2011. MicroRNA-7 inhibits the growth of human non-small cell lung cancer A549 cells through targeting BCL-2. Int J Biol Sci, 7(6):805-814.

[52]ZhangBY, WangO, QinJC, et al., 2013. cis-Acting elements and trans-acting factors in the transcriptional regulation of Raf kinase inhibitory protein expression. PLoS ONE, 8(12):e83097.

[53]ZhangEB, YinDD, SunM, et al., 2014. P53-regulated long non-coding RNA TUG1 affects cell proliferation in human non-small cell lung cancer, partly through epigenetically regulating HOXB7 expression. Cell Death Dis, 5(5):e1243.

[54]ZhangR, WangQ, ZhangL, et al., 2015. Optimized human factor IX expression cassettes for hepatic-directed gene therapy of hemophilia B. Front Med, 9(1):90-99.

[55]ZhaoJJ, WangKL, LiaoZY, et al., 2015. Promoter mutation of tumor suppressor microRNA-7 is associated with poor prognosis of lung cancer. Mol Clin Oncol, 3(6):‍1329-1336.

[56]ZhaoJJ, ChuFY, XuHL, et al., 2021. C/EBPα/miR-7 controls CD4+ T-cell activation and function and orchestrates experimental autoimmune hepatitis in mice. Hepatology, 74(1):379-396.

[57]ZhaoX, YangJ, HuangRY, et al., 2021. The role and its mechanism of intermittent fasting in tumors: friend or foe? Cancer Biol Med, 18(1):63-73.

[58]ZhuangXB, ZhaoC, LiJY, et al., 2019. Clinical features and therapeutic options in non-small cell lung cancer patients with concomitant mutations of EGFR, ALK, ROS1, KRAS or BRAF. Cancer Med, 8(6):2858-2866.

Open peer comments: Debate/Discuss/Question/Opinion


Please provide your name, email address and a comment

Journal of Zhejiang University-SCIENCE, 38 Zheda Road, Hangzhou 310027, China
Tel: +86-571-87952783; E-mail: cjzhang@zju.edu.cn
Copyright © 2000 - 2024 Journal of Zhejiang University-SCIENCE