Gene Expression Analysis of Interleukin 8 in NSCLC Cell Lines and Preparation of Noncovalently Functionalized Single Walled Carbon Nanotubes Conjugated with IL 8 siRNA
Abstract
Interleukin-8/CXCL 8 (IL-8) is a potent chemokine, angiogenic factor, autocrine growth factor with direct growth-promoting effect on many human cancers, and overexpression of IL-8 in carcinomas allows tumor growth and invasion. Ribonucleic acid (RNA) interference is an effective post-transcriptional gene silencing concept, specifically identifying and down-regulating the target gene’s expression. Small interfering RNAs (siRNAs) acts as a promising and powerful tool for application in gene therapy. However, siRNA therapies have been impeded by a lack of potential delivery systems. In this scenario, carbon nanotubes are used as an emerging platform for constructing safe, specific, and effective siRNA delivery systems. In this present work, the gene expression of IL-8 was studied in A 549 and H 460 non-small cell lung cancer cells using reverse transcription polymerase chain reaction (RT-PCR), and enzyme-linked immune sorbent assay produced significant levels of IL 8 protein (9.57 ± 1.6 ng mL-1 per 106cells and 83.26 ± 2.8 ng mL-1 per 106cells). Then potential tumor targeting nano delivery system was designed using single-walled carbon nanotubes (SWNTs), non-covalently functionalized with DSPE-PEG 2000 amine polymer to improve the solubility biocompatibility and lower the cytotoxicity. Then these functionalized SWNTs were conjugated to IL 8 siRNA via cleavable disulfide bond using a Sulfo-LC-SPDP cross-linker. The optimized formulation with enhanced biocompatibility and solubility may effectively be used as a nanocarrier for the potential delivery of IL 8 siRNA to tumor target sites.
Keywords:
Interleukin 8, Biocompatibility, Gene expression, small interfering RNADOI
https://doi.org/10.25004/IJPSDR.2021.130214References
Selma Rivas-Fuentes, Alfonso Salgado-Aguayo, Silvana Pertuz Belloso, Patricia Gorocica Rosete, Noé Alvarado-Vásquez, and Guillermo Aquino-Jarquin. Role of Chemokines in Non-Small Cell Lung Cancer: Angiogenesis and Inflammation. J Cancer. 2015; 6(10): 938–952.
Justin M. David, Charli Dominguez, Duane H. Hamilton, and Claudia Palena. The IL-8/IL-8R Axis: A Double Agent in Tumor Immune Resistance. Vaccines (Basel). 2016; 4(3):22.
Xinxin Long, Yingnan Ye, Lijie Zhang, Pengpeng Liu, Wenwen Yu, Feng Wei, Xiubao Ren and Jinpu Yu. IL-8, A novel messenger to cross-link inflammation and tumor EMT via autocrine and paracrine pathways. International Journal of Oncology. 2016;48(1):5-12.
M. F. Sanmamed J. L. Perez-Gracia K. A. Schalper J. P. Fusco. Changes in serum interleukin-8 (IL-8) levels reflect and predict response to anti-PD-1 treatment in melanoma and non-small-cell lung cancer patients. Annals of Oncology. 2017;28(8):1988-1995.
Boudreau R L and Davidson B L. RNAi therapeutics for CNS disorders. Brain Res. 2010;1338:112-21.
Suckau L et al. Long-term cardiac-targeted RNA interference for the treatment of heart failure restores cardiac function and reduces pathological hypertrophy. Circulation. 2009; 119 (9):1241-52.
Robert E. Farrell. RNAi: Take a RISC–Role the Dicer. RNA Methodologies. Edn 4, Elsevier, 2010, pp. 539-560.
Peixuan Guo, Oana Coban, Nick Snead, Joe Trebley, Steve Hoeprich, Songchuan Guo, and Yi Shu. Engineering RNA for Targeted siRNA Delivery and Medical Application. Adv Drug Deliv Rev. 2010; 62(6): 650–666.
Cheung,W, Pontoriero, F, Taratula,O ,Chen A.M. DNA and carbon nanotubes as medicine. Adv. Drug Deliv. Rev. 2010; 62: 633–649.
Shadpour Mal lakpour and Samaneh Soltanian. Surface functionalization of carbon nanotubes: fabrication and applications. RSC Advances. 2016; 6: 109916 –109935.
King Sun Siu et al. Non-covalently functionalized single-walled carbon nanotube for topical siRNA delivery into melanoma. Biomaterials. 2014; 35(10): 3435-3442.
Liu Z, Winters M, Holodniy, M,Dai, H. siRNA delivery into human Tcells and primary cells with carbon-nanotube transporters. Angew. Chem. Int. Ed. Engl. 2007; 46: 2023-2027.
Zhuang Liu, Scott Tabakman, Zhuo Chen, Hongjie Dai. Preparation of carbon nanotube bioconjugates for biomedical applications. Nat Protoc. 2009; 4(9):1372-82.
Wang J, Huang M, Lee P, Komanduri K, Sharma S, Chen G, Dubinett SM. Interleukin-8 inhibits non-small cell lung cancer proliferation: a possible role for regulation of tumor growth by autocrine and paracrine pathways. J Interferon Cytokine Res. 1996; 16: 53–60.
Arenberg DA, Kunkel SL, Polverini PJ, Glass M, Burdick MD, Strieter RM. Inhibition of interleukin-8 reduces tumorigenesis of human non-small cell lung cancer in SCID mice. J Clin Invest. 1996; 97: 2792-2802.
Noriaki Sunaga, Hisao Imai, Kimihiro Shimizu, David S Shames, Seiichi Kakegawa, Luc Girard, Mitsuo Sato, Kyoichi Kaira, Tamotsu Ishizuka, Adi F Gazdar, John D Minna, Masatomo Mori. Oncogenic KRAS-induced interleukin-8 overexpression promotes cell growth and migration and contributes to aggressive phenotypes of non-small cell lung cancer. Int J Cancer. 2012; 130(8):1733-44.
Chen JJW, Yao PL, Yuan A, Hong T-M, Shun C-T, Kuo M-L, Lee Y-C, Yang P-. Up-regulation of tumor interleukin-8 expression by infiltrating macrophages: its correlation with tumor angiogenesis and patient survival in non-small cell lung cancer. Clin Cancer Res. 2003; 9: 729-737.
Yuan-Pin Huang, I-Jou Lin, Chih-Chen Chen, Yi-Chiang Hsu, Chi- Chang Chang, Mon-Juan Lee. Delivery of small interfering RNAs in human cervical cancer cells by polyethylenimine-functionalized carbon nanotubes. Nanoscale Research Letters. 2013; 8(1):267.
Podesta JE, Al-Jamal KT, Herrero MA, et al. Antitumor activity and prolonged survival by carbon nanotube mediated therapeutic siRNA silencing in a human lung xenograft model. Small. 2009; 5:1176-85.
Haixia Li, Nan Zhang, Yongwei Hao, Yali Wang, Shasha Jia, Hongling Zhang, Yun Zhang, and Zhenzhong Zhang. Formulation of curcumin delivery with functionalized single-walled carbon nanotubes: characteristics and anticancer effects in vitro. Drug Deliv. 2014; 21(5):379-87.
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