Lidocaine inhibits the progression of retinoblastoma in vitro and in vivo by modulating the miR‑520a‑3p/EGFR axis

Xia,Weiyi; Wang,Libo; Yu,Dongyi; Mu,Xing; Zhou,Xin

doi:10.3892/mmr.2019.10363

Lidocaine inhibits the progression of retinoblastoma in vitro and in vivo by modulating the miR‑520a‑3p/EGFR axis

Authors:
- Weiyi Xia
- Libo Wang
- Dongyi Yu
- Xing Mu
- Xin Zhou
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Affiliations: Department of Ophthalmology, The First People's Hospital of Kunshan Affiliated with Jiangsu University, Suzhou, Jiangsu 215300, P.R. China
Published online on: June 6, 2019 https://doi.org/10.3892/mmr.2019.10363
Pages: 1333-1342
Copyright: © Xia et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Retinoblastoma (RB) is a common malignant tumor in children. Lidocaine is a local anesthetic and anti‑arrhythmic drug, and has been reported to possess anti‑tumor properties. MicroRNAs (miRs) are a group of endogenous small noncoding RNAs that have important roles in various biological processes via actions on target genes. The aim of the present study was to investigate the effect of lidocaine on retinoblastoma in vitro and in vivo. CCK‑8 assay and flow cytometry assay were used to measure cell viability and apoptosis. The relationship between miR‑520a‑3p and EGFR was predicted and confirmed by TargetScan and dual‑luciferase reporter assay. For in vivo study, tumor xenograft was performed. In addition, gene and protein expression was detected using reverse transcription‑quantitative polymerase chain reaction and western blotting respectively. In the present study, it was observed that lidocaine inhibited the proliferation and induced the apoptosis of RB cells. miR‑520a‑3p was reported to be downregulated in RB tissues and cell lines; treatment with lidocaine increased the expression of miR‑520a‑3p in RB cells. The human epidermal growth factor receptor (EGFR) was identified as a direct target of miR‑520a‑3p, and its expression was negatively associated with that of miR‑520a‑3p. Additionally, EGFR was upregulated in RB tissues and cell lines; treatment with lidocaine decreased the expression of EGFR in RB cells. Furthermore, compared with treatment with lidocaine alone, the combination of transfection with miR‑520a‑3p inhibitor and lidocaine treatment significantly decreased the expression of miR‑520a‑3p, increased EGFR expression, promoted RB cell proliferation and reduced the apoptosis of cells in vitro, and increased tumor volume and weight in vivo. The results indicated that lidocaine reduced the proliferation and induced the apoptosis of RB cells by decreasing EGFR expression via the upregulation of miR‑520a‑3p, suggesting that the miR‑520a‑3p/EGFR axis may be a novel therapeutic target in the treatment of RB.

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1	Ortiz MV and Dunkel IJ: Retinoblastoma. J Child Neurol. 31:227–236. 2016. View Article : Google Scholar : PubMed/NCBI
2	Rao R and Honavar SG: Retinoblastoma. Indian J Pediatr. 84:937–944. 2017. View Article : Google Scholar : PubMed/NCBI
3	Chen BJ, Mariño-Enríquez A, Fletcher CD and Hornick JL: Loss of retinoblastoma protein expression in spindle cell/pleomorphic lipomas and cytogenetically related tumors: An immunohistochemical study with diagnostic implications. Am J Surg Pathol. 36:1119–1128. 2012. View Article : Google Scholar : PubMed/NCBI
4	Tetzlaff JE: The pharmacology of local anesthetics. Anesthesiol Clin North Am. 18:217–233. 2000. View Article : Google Scholar : PubMed/NCBI
5	Caracas HC, Maciel JV, Martins PM, de Souza MM and Maia LC: The use of lidocaine as an anti-inflammatory substance: A systematic review. J Dent. 37:93–97. 2009. View Article : Google Scholar : PubMed/NCBI
6	Adler DMT, Damborg P and Verwilghen DR: The antimicrobial activity of bupivacaine, lidocaine and mepivacaine against equine pathogens: An investigation of 40 bacterial isolates. Vet J. 223:27–31. 2017. View Article : Google Scholar : PubMed/NCBI
7	Gao X, Yang H, Wu M, Shi K, Zhou C, Peng J and Yang Q: Targeting delivery of lidocaine and cisplatin by nanogel enhances chemotherapy and alleviates metastasis. ACS Appl Mater Interfaces. 10:25228–25240. 2018. View Article : Google Scholar : PubMed/NCBI
8	Johnson MZ, Crowley PD, Foley AG, Xue C, Connolly C, Gallagher HC and Buggy DJ: Effect of perioperative lidocaine on metastasis after sevoflurane or ketamine-xylazine anaesthesia for breast tumour resection in a murine model. Br J Anaesth. 121:76–85. 2018. View Article : Google Scholar : PubMed/NCBI
9	Calin GA and Croce CM: MicroRNA-cancer connection: The beginning of a new tale. Cancer Res. 66:7390–7394. 2006. View Article : Google Scholar : PubMed/NCBI
10	Esquela-Kerscher A and Slack FJ: Oncomirs-microRNAs with a role in cancer. Nat Rev Cancer. 6:259–269. 2006. View Article : Google Scholar : PubMed/NCBI
11	McManus MT: MicroRNAs and cancer. Semin Cancer Biol. 13:253–258. 2003. View Article : Google Scholar : PubMed/NCBI
12	Ro S, Park C, Young D, Sanders KM and Yan W: Tissue-dependent paired expression of miRNAs. Nucleic Acids Res. 35:5944–5953. 2007. View Article : Google Scholar : PubMed/NCBI
13	Mallory AC and Vaucheret H: MicroRNAs: Something important between the genes. Curr Opin Plant Biol. 7:120–125. 2004. View Article : Google Scholar : PubMed/NCBI
14	Garzon R, Calin GA and Croce CM: MicroRNAs in cancer. Annu Rev Med. 60:167–179. 2009. View Article : Google Scholar : PubMed/NCBI
15	Wu N, Song Y, Pang L and Chen Z: CRCT1 regulated by microRNA-520 g inhibits proliferation and induces apoptosis in esophageal squamous cell cancer. Tumor Biol. 37:8271–8279. 2016. View Article : Google Scholar
16	Keklikoglou I, Koerner C, Schmidt C, Zhang JD, Heckmann D, Shavinskaya A, Allgayer H, Gückel B, Fehm T, Schneeweiss A, et al: MicroRNA-520/373 family functions as a tumor suppressor in estrogen receptor negative breast cancer by targeting NF-κB and TGF-β signaling pathways. Oncogene. 31:4150–4163. 2012. View Article : Google Scholar : PubMed/NCBI
17	Ding M, Lu X, Wang C, Zhao Q, Ge J, Xia Q, Wang J, Zen K, Zhang CY and Zhang C: The E2F1-miR-520/372/373-SPOP axis modulates progression of renal carcinoma. Cancer Res. 78:6771–6784. 2018. View Article : Google Scholar : PubMed/NCBI
18	Park YY, Kim SB, Han HD, Sohn BH, Kim JH, Liang J, Lu Y, Rodriguez-Aguayo C, Lopez-Berestein G, Mills GB, et al: Tat-activating regulatory DNA-binding protein regulates glycolysis in hepatocellular carcinoma by regulating the platelet isoform of phosphofructokinase through microRNA 520. Hepatology. 58:182–191. 2013. View Article : Google Scholar : PubMed/NCBI
19	Zhang R, Liu R, Liu C, Niu Y, Zhang J, Guo B, Zhang CY, Li J, Yang J and Chen X: A novel role for MiR-520a-3p in regulating EGFR expression in colorectal cancer. Cell Physiol Biochem. 42:1559–1574. 2017. View Article : Google Scholar : PubMed/NCBI
20	Liu Y, Miao L, Ni R, Zhang H, Li L, Wang X, Li X and Wang J: microRNA-520a-3p inhibits proliferation and cancer stem cell phenotype by targeting HOXD8 in non-small cell lung cancer. Oncol Rep. 36:3529–3535. 2016. View Article : Google Scholar : PubMed/NCBI
21	Chen A, Xu J and Johnson AC: Curcumin inhibits human colon cancer cell growth by suppressing gene expression of epidermal growth factor receptor through reducing the activity of the transcription factor Egr-1. Oncogene. 25:278–287. 2006. View Article : Google Scholar : PubMed/NCBI
22	Cîrstea AE, Stepan AE, Zăvoi RE and Simionescu CE: EGFR Immunoexpression in malignant serous and mucinous ovarian tumors. Curr Health Sci J. 44:129–134. 2018.PubMed/NCBI
23	Jia Y, Li X, Jiang T, Zhao S, Zhao C, Zhang L, Liu X, Shi J, Qiao M, Luo J, et al: EGFR-targeted therapy alters the tumor microenvironment in EGFR-driven lung tumors: Implications for combination therapies. Int J Cancer. Feb 5–2019.doi: 10.1002/ijc.32191 (Epub ahead of print). View Article : Google Scholar
24	Wang P, Liu X, Shao Y, Wang H, Liang C, Han B and Ma Z: MicroRNA-107-5p suppresses non-small cell lung cancer by directly targeting oncogene epidermal growth factor receptor. Oncotarget. 8:57012–57023. 2017.PubMed/NCBI
25	Sakaguchi M, Kuroda Y and Hirose M: The antiproliferative effect of lidocaine on human tongue cancer cells with inhibition of the activity of epidermal growth factor receptor. Anesth Analg. 102:1103–1107. 2006. View Article : Google Scholar : PubMed/NCBI
26	Bundscherer AC, Malsy M, Bitzinger DI, Wiese CH, Gruber MA and Graf BM: Effects of lidocaine on HT-29 and SW480 colon cancer cells in vitro. Anticancer Res. 37:1941–1945. 2017. View Article : Google Scholar : PubMed/NCBI
27	Qu X, Yang L, Shi Q, Wang X, Wang D and Wu G: Lidocaine inhibits proliferation and induces apoptosis in colorectal cancer cells by upregulating mir-520a-3p and targeting EGFR. Pathol Res Pract. 214:1974–1979. 2018. View Article : Google Scholar : PubMed/NCBI
28	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
29	Bayne K: Revised guide for the care and use of laboratory animals available. American physiological society. Physiologist. 39:199, 208–211. 1996.
30	Freeman J, Crowley PD, Foley AG, Gallagher HC, Iwasaki M, Ma D and Buggy DJ: Effect of perioperative lidocaine and cisplatin on metastasis in a murine model of breast cancer surgery. Anticancer Res. 38:5599–5606. 2018. View Article : Google Scholar : PubMed/NCBI
31	Zhang Y, Zhu X, Zhu X, Wu Y, Liu Y, Yao B and Huang Z: MiR-613 suppresses retinoblastoma cell proliferation, invasion, and tumor formation by targeting E2F5. Tumour Biol. 39:10104283176916742017.PubMed/NCBI
32	Di LG and Croce CM: miRNA profiling of cancer. Curr Opin Genet Dev. 23:3–11. 2013. View Article : Google Scholar : PubMed/NCBI
33	Wei W, Yang Y, Cai J, Cui K, Li RX, Wang H, Shang X and Wei D: MiR-30a-5p Suppresses tumor metastasis of human colorectal cancer by targeting ITGB3. Cell Physiol Biochem. 39:1165–1176. 2016. View Article : Google Scholar : PubMed/NCBI
34	Liu X, Xie T, Mao X, Xue L, Chu X and Chen L: MicroRNA-149 increases the sensitivity of colorectal cancer cells to 5-fluorouracil by targeting forkhead box transcription factor FOXM1. Cell Physiol Biochem. 39:617–629. 2016. View Article : Google Scholar : PubMed/NCBI
35	Zhou L, Xu Z, Ren X, Chen K and Xin S: MicroRNA-124 (MiR-124) inhibits cell proliferation, metastasis and invasion in colorectal cancer by downregulating Rho-associated protein kinase 1(ROCK1). Cell Physiol Biochem. 38:1785–1795. 2016. View Article : Google Scholar : PubMed/NCBI
36	Li HT, Zhang H, Chen Y, Liu XF and Qian J: MiR-423-3p enhances cell growth through inhibition of p21Cip1/Waf1 in colorectal cancer. Cell Physiol Biochem. 37:1044–1054. 2015. View Article : Google Scholar : PubMed/NCBI
37	Nishimura M, Jung EJ, Shah MY, Lu C, Spizzo R, Shimizu M, Han HD, Ivan C, Rossi S, Zhang X, et al: Therapeutic synergy between microRNA and siRNA in ovarian cancer treatment. Cancer Discov. 3:1302–1315. 2013. View Article : Google Scholar : PubMed/NCBI
38	Li R, Yuan W, Mei W, Yang K and Chen Z: MicroRNA 520d-3p inhibits gastric cancer cell proliferation, migration, and invasion by downregulating EphA2 expression. Mol Cell Biochem. 396:295–305. 2014. View Article : Google Scholar : PubMed/NCBI
39	Gach K, Wyrębska A, Fichna J and Janecka A: The role of morphine in regulation of cancer cell growth. Naunyn Schmiedebergs Arch Pharmacol. 384:221–230. 2011. View Article : Google Scholar : PubMed/NCBI
40	Xu YB, Du QH, Zhang MY, Yun P and He CY: Propofol suppresses proliferation, invasion and angiogenesis by down-regulating ERK-VEGF/MMP-9 signaling in Eca-109 esophageal squamous cell carcinoma cells. Eur Rev Med Pharmacol Sci. 17:2486–2494. 2013.PubMed/NCBI
41	Chang YC, Liu CL, Chen MJ, Hsu YW, Chen SN, Lin CH, Chen CM, Yang FM and Hu MC: Local anesthetics induce apoptosis in human breast tumor cells. Anesth Analg. 118:116–124. 2014. View Article : Google Scholar : PubMed/NCBI
42	Yu J, Tan Q, Deng B, Fang C, Qi D and Wang R: The microRNA-520a-3p inhibits proliferation, apoptosis and metastasis by targeting MAP3K2 in non-small cell lung cancer. Am J Cancer Res. 5:802–811. 2015.PubMed/NCBI
43	Li J, Wei J, Mei Z, Yin Y, Li Y, Lu M and Jin S: Suppressing role of miR-520a-3p in breast cancer through CCND1 and CD44. Am J Transl Res. 9:146–154. 2017.PubMed/NCBI