Safety markers for rhabdomyosarcoma cells using an in vivo imaging system

Du,Shutong; Meng,Lian; Song,Lingxie; Zhang,Pengpeng; Shou,Xi; Liu,Chunxia; Li,Feng

doi:10.3892/ol.2018.8789

Safety markers for rhabdomyosarcoma cells using an in vivo imaging system

Authors:
- Shutong Du
- Lian Meng
- Lingxie Song
- Pengpeng Zhang
- Xi Shou
- Chunxia Liu
- Feng Li
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Affiliations: Department of Pathology, School of Medicine, Shihezi University, Shihezi, Xinjiang 832002, P.R. China, Department of Animal Experiment Center, Xinjiang Key Laboratory for Medical Animal Model Research, The First Affiliated Hospital of Xinjiang Medical University, Ürümqi, Xinjiang 830011, P.R. China
Published online on: May 22, 2018 https://doi.org/10.3892/ol.2018.8789
Pages: 1031-1038

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Abstract

In vivo imaging system (IVIS) is a novel and rapidly expanding technology that is widely applied in life sciences, including cell tracing. IVIS is able to quantify biological events, including tumor proliferation, through counting the number of photons emitted from a specimen. PLA802‑enhanced green fluorescent protein (EGFP), PLA802‑monomeric cherry fluorescent protein (mCherry), RH30‑EGFP and RH30‑mCherry tumor cells were injected into 18 BALB/c female nude mice subcutaneously with 5x106 cells in 100 µl to quantitatively analyze EGFP and mCherry cells traced by IVIS. Inversion fluorescence microscopy revealed no transfection efficiency difference between PLA802‑EGFP (95.3±1.2%) and PLA802‑mCherry (95.8±1.7%), or between RH30‑EGFP (94.7±2.1%) and RH30‑mCherry (95.2±1.9%). Transfection did not influence the cell morphology of PLA802 or RH30. The cell migration, invasion and proliferation assay results of lentivirus‑EGFP and lentivirus‑mCherry revealed no significant difference prior to or following transfection. Therefore, lentivirus‑EGFP and lentivirus‑mCherry may serve as safety biological markers for PLA802 and RH30 cells. In vivo experiments demonstrated that lentivirus‑EGFP and lentivirus‑mCherry tumor luminescence signals were observed in all mice by IVIS. Hematoxylin‑eosin staining and immunohistochemistry indicated that PLA802‑EGFP, PLA802‑mCherry, RH30‑EGFP and RH30‑mCherry cell lines exhibited rhabdomyosarcoma (RMS) characteristics like the maternal cells. In summary, mCherry and green fluorescent protein in human RMS PLA802 and RH30 cancer cells may be safely and stably expressed for a long time in vitro and in vivo.

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1	Kashi VP, Hatley ME and Galindo RL: Probing for a deeper understanding of rhabdomyosarcoma: Insights from complementary model systems. Nat Rev Cancer. 15:426–439. 2015. View Article : Google Scholar : PubMed/NCBI
2	Parham DM and Ellison DA: Rhabdomyosarcomas in adults and children: An update. Arch Pathol Lab Med. 130:1454–1465. 2006.PubMed/NCBI
3	Zhang M, Linardic CM and Kirsch DG: RAS and ROS in rhabdomyosarcoma. Cancer Cell. 24:689–691. 2013. View Article : Google Scholar : PubMed/NCBI
4	Erratum: Inhibition of rhabdomyosarcoma cell and tumor growth by targeting specificity protein (sp) transcription factors. Int J Cancer. 137:E92015. View Article : Google Scholar : PubMed/NCBI
5	Hinson AR, Jones R, Crose LE, Belyea BC, Barr FG and Linardic CM: Human rhabdomyosarcoma cell lines for rhabdomyosarcoma research: Utility and pitfalls. Front Oncol. 3:1832013. View Article : Google Scholar : PubMed/NCBI
6	Ebe T: Green fluorescent protein as a marker gene expression. Tanpakushitsu Kakusan Koso. 52(Suppl 13): 1766–1767. 2007.(In Japanese). PubMed/NCBI
7	Zhang Q, Walawage SL, Tricoli DM, Dandekar AM and Leslie CA: A red fluorescent protein (DsRED) from discosoma sp. as a reporter for gene expression in walnut somatic embryos. Plant Cell Rep. 34:861–869. 2015.
8	Katoh A, Shoguchi K, Matsuoka H, Yoshimura M, Ohkubo JI, Matsuura T, Maruyama T, Ishikura T, Aritomi T, Fujihara H, et al: Fluorescent visualisation of the hypothalamic oxytocin neurones activated by cholecystokinin-8 in rats expressing c-fos-enhanced green fluorescent protein and oxytocin-monomeric red fluorescent protein 1 fusion transgenes. J Neuroendocrinol. 26:341–347. 2014. View Article : Google Scholar : PubMed/NCBI
9	Fink D, Wohrer S, Pfeffer M, Tombe T, Ong CJ and Sorensen PH: Ubiquitous expression of the monomeric red fluorescent protein mCherry in transgenic mice. Genesis. 48:723–729. 2010. View Article : Google Scholar : PubMed/NCBI
10	Rossetti M, Cavarelli M, Gregori S and Scarlatti G: HIV-derived vectors for gene therapy targeting dendritic cells. Adv Exp Med Biol. 762:239–261. 2013. View Article : Google Scholar : PubMed/NCBI
11	Chang LJ and He J: Retroviral vectors for gene therapy of AIDS and cancer. Curr Opin Mol Ther. 3:468–475. 2001.PubMed/NCBI
12	Shichinohe T, Bochner BH, Mizutani K, Nishida M, Hegerich-Gilliam S, Naldini L and Kasahara N: Development of lentiviral vectors for antiangiogenic gene delivery. Cancer Gene Ther. 8:879–889. 2001. View Article : Google Scholar : PubMed/NCBI
13	Dancsok AR, Asleh-Aburaya K and Nielsen TO: Advances in sarcoma diagnostics and treatment. Oncotarget. 8:7068–7093. 2017. View Article : Google Scholar : PubMed/NCBI
14	Shern JF, Yohe ME and Khan J: Pediatric rhabdomyosarcoma. Crit Rev Oncog. 20:227–243. 2015. View Article : Google Scholar : PubMed/NCBI
15	Casini N, Forte IM, Mastrogiovanni G, Pentimalli F, Angelucci A, Festuccia C, Tomei V, Ceccherini E, Di Marzo D, Schenone S, et al: SRC family kinase (SFK) inhibition reduces rhabdomyosarcoma cell growth in vitro and in vivo and triggers p38 MAP kinase-mediated differentiation. Oncotarget. 6:12421–12435. 2015. View Article : Google Scholar : PubMed/NCBI
16	Liu C, Li D, Jiang J, Hu J, Zhang W, Chen Y, Cui X, Qi Y, Zou H, Zhang W and Li F: Analysis of molecular cytogenetic alteration in rhabdomyosarcoma by array comparative genomic hybridization. PloS One. 9:e949242014. View Article : Google Scholar : PubMed/NCBI
17	Ntziachristos V, Ripoll J, Wang LV and Weissleder R: Looking and listening to light: The evolution of whole-body photonic imaging. Nat Biotechnol. 23:313–320. 2005. View Article : Google Scholar : PubMed/NCBI
18	Maggi A and Ciana P: Reporter mice and drug discovery and development. Nat Rev Drug Discov. 4:249–255. 2005. View Article : Google Scholar : PubMed/NCBI
19	Dubey P: Reporter gene imaging of immune responses to cancer: Progress and Challenges. Theranostics. 2:355–362. 2012. View Article : Google Scholar : PubMed/NCBI
20	Martelli C, Dico AL, Diceglie C, Lucignani G and Ottobrini L: Optical imaging probes in oncology. Oncotarget. 7:48753–48787. 2016. View Article : Google Scholar : PubMed/NCBI
21	Hoffman RM: The multiple uses of fluorescent proteins to visualize cancer in vivo. Nat Rev Cancer. 5:796–806. 2005. View Article : Google Scholar : PubMed/NCBI
22	Hoffman RM: Fluorescent proteins as visible in vivo sensors. Prog Mol Biol Transl Sci. 113:389–402. 2013. View Article : Google Scholar : PubMed/NCBI
23	Weissleder R: Molecular imaging: Exploring the next frontier. Radiology. 212:609–614. 1999. View Article : Google Scholar : PubMed/NCBI
24	Chen ZY, Wang YX, Lin Y, Zhang JS, Yang F, Zhou QL and Liao YY: Advance of molecular imaging technology and targeted imaging agent in imaging and therapy. Biomed Res Int. 2014:8193242014.PubMed/NCBI
25	Liu K, Wang MW, Lin WY, Phung DL, Girgis MD, Wu AM, Tomlinson JS and Shen CK: Molecular imaging probe development using microfluidics. Curr Org Synth. 8:473–487. 2011. View Article : Google Scholar : PubMed/NCBI
26	Hillman EM, Amoozegar CB, Wang T, McCaslin AF, Bouchard MB, Mansfield J and Levenson RM: In vivo optical imaging and dynamic contrast methods for biomedical research. Philos Trans A Math Phys Eng Sci. 369:4620–4643. 2011. View Article : Google Scholar : PubMed/NCBI
27	Chalfie M, Tu Y, Euskirchen G, Ward WW and Prasher DC: Green fluorescent protein as a marker for gene expression. Science. 263:802–805. 1994. View Article : Google Scholar : PubMed/NCBI
28	Adusumilli PS, Eisenberg DP, Stiles BM, Chung S, Chan MK, Rusch VW and Fong Y: Intraoperative localization of lymph node metastases with a replication-competent herpes simplex virus. J Thorac Cardiovasc Surg. 132:1179–1188. 2006. View Article : Google Scholar : PubMed/NCBI
29	Shen Y, Chen Y, Wu J, Shaner NC and Campbell RE: Engineering of mCherry variants with long Stokes shift, red-shifted fluorescence, and low cytotoxicity. PLoS One. 12:e01712572017. View Article : Google Scholar : PubMed/NCBI
30	Shen Y, Lai T and Campbell RE: Red fluorescent proteins (RFPs) and RFP-based biosensors for neuronal imaging applications. Neurophotonics. 2:0312032015. View Article : Google Scholar : PubMed/NCBI
31	Piatkevich KD and Verkhusha VV: Advances in engineering of fluorescent proteins and photoactivatable proteins with red emission. Curr Opin Chem Biol. 14:23–29. 2010. View Article : Google Scholar : PubMed/NCBI
32	Campbell RE, Tour O, Palmer AE, Steinbach PA, Baird GS, Zacharias DA and Tsien RY: A monomeric red fluorescent protein. Proc Natl Acad Sci USA. 99:7877–7882. 2002. View Article : Google Scholar : PubMed/NCBI
33	Merzlyak EM, Goedhart J, Shcherbo D, Bulina ME, Shcheglov AS, Fradkov AF, Gaintzeva A, Lukyanov KA, Lukyanov S, Gadella TW and Chudakov DM: Bright monomeric red fluorescent protein with an extended fluorescence lifetime. Nat Methods. 4:555–557. 2007. View Article : Google Scholar : PubMed/NCBI
34	Shaner NC, Campbell RE, Steinbach PA, Giepmans BN, Palmer AE and Tsien RY: Improved monomeric red, orange and yellow fluorescent proteins derived from discosoma sp. red fluorescent protein. Nat Biotechnol. 22:1567–1572. 2004. View Article : Google Scholar
35	Merten OW, Hebben M and Bovolenta C: Production of lentiviral vectors. Mol Ther Methods Clin Dev. 3:160172016. View Article : Google Scholar : PubMed/NCBI
36	Rashidian M, Dozier JK and Distefano MD: Chemoenzymatic labeling of proteins: Techniques and approaches. Bioconjug Chem. 24:1277–1294. 2013. View Article : Google Scholar : PubMed/NCBI
37	Lee K, Majumdar MK, Buyaner D, Hendricks JK, Pittenger MF and Mosca JD: Human mesenchymal stem cells maintain transgene expression during expansion and differentiation. Mol Ther. 3:857–866. 2001. View Article : Google Scholar : PubMed/NCBI