Lentiviral Vector with a Radiation-inducible Promoter, Carrying the ING4 Gene, Mediates Radiosensitization Controlled by Radiotherapy in Cervical Cancer Cells

Overview

Journal Oncol Lett

Specialty Oncology

Date 2020 Dec 28

PMID 33365078

Authors

Tao Ma

Rui Guo

Xi Wang

Wen-Tong Shen

Min Zhu

Ye-Ning Jin

Hao-Ping Xu

Affiliations

Soon will be listed here.

Abstract

The presence of hypoxia in solid tumors is considered one of the major factors that contribute to radiation resistance. The aim of the present study was to establish a therapeutic system, which can be controlled by radiation itself, to enhance radiosensitivity. For this purpose, a lentiviral gene therapy vector containing the human inhibitor of growth 4 (ING4) and its upstream promoter, human early growth response factor-1 (EGR1), which possesses the radiation-inducible characteristics to activate the transcription of its downstream genes, was constructed. Downstream fluorescence proteins were investigated to ensure that the EGR1 promoter was induced by irradiation. Furthermore, ING4 open reading frame (ORF) expression was detected by western blotting. The cell cycle was analyzed by fluorescence-activated cell sorting analysis 48 h after the cells were exposed to X-rays ranging between 0 and 8 Gy. In cells stably and transiently transfected with reporter plasmids, the EGR1-driver gene was sensitive to ionizing irradiation. Furthermore, irradiation-induced ING4 gene expression was observed. The enhanced ING4 expression increased the number of cells in the G/M phase and decreased the proportion of cells in the G/S phase. Therefore, ING4 expression inhibited cell proliferation and was associated with less colonies being formed. Furthermore, ING4 suppressed hypoxia-inducible factor 1α expression under hypoxic conditions and promoted cell apoptosis. Overall, these results revealed that combining the EGR1 promoter and ING4 ORF using a lentivirus system may be a promising therapeutic strategy with which to enhance radiosensitivity controlled by radiation. However, further studies using models are required to confirm these findings.

References

Rockwell S, Dobrucki I, Kim E, Marrison S, Vu V . Hypoxia and radiation therapy: past history, ongoing research, and future promise. Curr Mol Med. 2009; 9(4):442-58. PMC: 2752413. DOI: 10.2174/156652409788167087. View

Chino J, Annunziata C, Beriwal S, Bradfield L, Erickson B, Fields E . Radiation Therapy for Cervical Cancer: Executive Summary of an ASTRO Clinical Practice Guideline. Pract Radiat Oncol. 2020; 10(4):220-234. PMC: 8802172. DOI: 10.1016/j.prro.2020.04.002. View

Toma-Dasu I, Dasu A, Karlsson M . The relationship between temporal variation of hypoxia, polarographic measurements and predictions of tumour response to radiation. Phys Med Biol. 2004; 49(19):4463-75. DOI: 10.1088/0031-9155/49/19/002. View

Du Y, Cheng Y, Su G . The essential role of tumor suppressor gene in various human cancers and non-neoplastic disorders. Biosci Rep. 2019; 39(1). PMC: 6356015. DOI: 10.1042/BSR20180773. View

Yang W, Li X . Anti-tumor effect of pEgr-interferon-gamma-endostatin gene-radiotherapy in mice bearing Lewis lung carcinoma and its mechanism. Chin Med J (Engl). 2005; 118(4):296-301. View

Leskov K, Criswell T, Antonio S, Li J, Yang C, Kinsella T . When X-ray-inducible proteins meet DNA double strand break repair. Semin Radiat Oncol. 2001; 11(4):352-72. DOI: 10.1053/srao.2001.26912. View

You Q, Wang X, Fu S, Jin X . Downregulated expression of inhibitor of growth 4 (ING4) in advanced colorectal cancers: a non-randomized experimental study. Pathol Oncol Res. 2011; 17(3):473-7. DOI: 10.1007/s12253-010-9301-7. View

Lukas J, Lukas C, Bartek J . Mammalian cell cycle checkpoints: signalling pathways and their organization in space and time. DNA Repair (Amst). 2004; 3(8-9):997-1007. DOI: 10.1016/j.dnarep.2004.03.006. View

Wang Q, Li M, Zhang L, Jin Y, Tong D, Yu Y . Down-regulation of ING4 is associated with initiation and progression of lung cancer. Histopathology. 2010; 57(2):271-81. PMC: 3855453. DOI: 10.1111/j.1365-2559.2010.03623.x. View

10.

Wang Y, Yang J, Sheng W, Xie Y, Liu J . Adenovirus-mediated ING4/PTEN double tumor suppressor gene co-transfer modified by RGD enhances antitumor activity in human nasopharyngeal carcinoma cells. Int J Oncol. 2015; 46(3):1295-303. DOI: 10.3892/ijo.2015.2822. View

11.

Hickey M, Simon M . Regulation of angiogenesis by hypoxia and hypoxia-inducible factors. Curr Top Dev Biol. 2006; 76:217-57. DOI: 10.1016/S0070-2153(06)76007-0. View

12.

Moeller B, Dewhirst M . HIF-1 and tumour radiosensitivity. Br J Cancer. 2006; 95(1):1-5. PMC: 2360497. DOI: 10.1038/sj.bjc.6603201. View

13.

Zhang H, Zhou X, Xu C, Yang J, Xiang J, Tao M . Synergistic tumor suppression by adenovirus-mediated ING4/PTEN double gene therapy for gastric cancer. Cancer Gene Ther. 2015; 23(1):13-23. DOI: 10.1038/cgt.2015.59. View

14.

Rovetta F, Stacchiotti A, Faggi F, Catalani S, Apostoli P, Fanzani A . Cobalt triggers necrotic cell death and atrophy in skeletal C2C12 myotubes. Toxicol Appl Pharmacol. 2013; 271(2):196-205. DOI: 10.1016/j.taap.2013.05.005. View

15.

Yan R, He L, Li Z, Han X, Liang J, Si W . SCF(JFK) is a bona fide E3 ligase for ING4 and a potent promoter of the angiogenesis and metastasis of breast cancer. Genes Dev. 2015; 29(6):672-85. PMC: 4378198. DOI: 10.1101/gad.254292.114. View

16.

Liu L, Smith M, Sun B, Wang G, Redmond H, Wang J . Combined IFN-gamma-endostatin gene therapy and radiotherapy attenuates primary breast tumor growth and lung metastases via enhanced CTL and NK cell activation and attenuated tumor angiogenesis in a murine model. Ann Surg Oncol. 2009; 16(5):1403-11. DOI: 10.1245/s10434-009-0343-6. View

17.

Weichselbaum R, Kufe D . Translation of the radio- and chemo-inducible TNFerade vector to the treatment of human cancers. Cancer Gene Ther. 2009; 16(8):609-19. DOI: 10.1038/cgt.2009.37. View

18.

Zhao Y, Li Z, Sheng W, Miao J, Yang J . Radiosensitivity by ING4-IL-24 bicistronic adenovirus-mediated gene cotransfer on human breast cancer cells. Cancer Gene Ther. 2012; 20(1):38-45. DOI: 10.1038/cgt.2012.82. View

19.

Xie Y, Zhang H, Sheng W, Xiang J, Ye Z, Yang J . Adenovirus-mediated ING4 expression suppresses lung carcinoma cell growth via induction of cell cycle alteration and apoptosis and inhibition of tumor invasion and angiogenesis. Cancer Lett. 2008; 271(1):105-16. DOI: 10.1016/j.canlet.2008.05.050. View

20.

Galal El-Shemi A, Mohammed Ashshi A, Oh E, Jung B, Basalamah M, Alsaegh A . Efficacy of combining ING4 and TRAIL genes in cancer-targeting gene virotherapy strategy: first evidence in preclinical hepatocellular carcinoma. Gene Ther. 2017; 25(1):54-65. PMC: 5817393. DOI: 10.1038/gt.2017.86. View