Antimicrobial peptide 17BIPHE2 inhibits the proliferation of lung cancer cells <em>in vitro</em> and <em>in vivo</em> by regulating the ERK signaling pathway

Yang,Tingting; Li,Jun; Jia,Qinqin; Zhan,Shisheng; Zhang,Qiannan; Wang,Yarong; Wang,Xiuqing

doi:10.3892/ol.2021.12762

Antimicrobial peptide 17BIPHE2 inhibits the proliferation of lung cancer cells in vitro and in vivo by regulating the ERK signaling pathway

Authors:
- Tingting Yang
- Jun Li
- Qinqin Jia
- Shisheng Zhan
- Qiannan Zhang
- Yarong Wang
- Xiuqing Wang
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Affiliations: Department of Clinical Laboratory, Yinchuan Maternal and Child Health Care Hospital, Yinchuan, Ningxia Hui Autonomous Region 750004, P.R. China, Department of Clinical Laboratory, Tangshan Gongren Hospital, Tangshan, Hebei 063000, P.R. China, Department of Laboratory Medicine, Health Center, Chun Rong, Gansu 745211, P.R. China, Department of Clinical Laboratory, Hebei Yanda Lu Daopei Hospital, Langfang, Hebei 065200, P.R. China, Department of Laboratory Medicine, College of Clinical Medicine, Shuangyi Campus, Ningxia Medical University, Yinchuan, Ningxia Hui Autonomous Region 750004, P.R. China, Department of Pathology, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences, Tianjin 300020, P.R. China
Published online on: April 28, 2021 https://doi.org/10.3892/ol.2021.12762
Article Number: 501
Copyright: © Yang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

In 2018, there were 18.1 million new cancer cases and 9.6 million cancer‑related deaths worldwide, among which the incidence rate of lung cancer (11.6%) and fatality rate (18.4%) both ranked first. The antimicrobial peptide LL‑37 is an important component of the natural immune system and possesses several biological properties, including antibacterial, antiviral and anticancer effects. The antimicrobial peptide 17BIPHE2, the shortest synthetic peptide derivative of LL‑37, exhibits biological activities similar to those of LL‑37. The objective of the present study was to investigate the mechanism of action of exogenous 17BIPHE2 against lung cancer cells. The human lung adenocarcinoma cell line A549 was treated with 17BIPHE2. Changes in cell proliferation, migration, invasion, mitochondrial membrane potential (ΔΨm), and the levels of reactive oxygen species (ROS), Ca²⁺ and apoptosis‑related proteins, including BAX, BCL‑2 and ERK, were detected using flow cytometry, transmission electron microscopy and western blotting. The results showed that 17BIPHE2 significantly increased the apoptosis rate of A549 cells and elevated BAX expression, ERK phosphorylation, and ROS and Ca²⁺ levels, but decreased the expression of BCL‑2, ERK and Ki67. In addition, the peptide reduced ΔΨm and the cell migration ability of A549 cells and inhibited tumor growth. ERK inhibition significantly attenuated the anticancer effect of 17BIPHE2. The present observations suggested that 17BIPHE2 can effectively inhibit cancer cells by regulating the ERK signaling pathway.

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1	Singh RD, Shandilya R, Bhargava A, Kumar R, Tiwari R, Chaudhury K, Srivastava RK, Goryacheva IY and Mishra PK: Quantum dot based nano-biosensors for detection of circulating cell free miRNAs in lung carcinogenesis: From Biology to Clinical translation. Front Genet. 9:6162018. View Article : Google Scholar : PubMed/NCBI
2	Chen W, Zheng R, Baade PD, Zhang S, Zeng H, Bray F, Jemal A, Yu XQ and He J: Cancer statistics in China, 2015. CA Cancer J Clin. 66:115–132. 2016. View Article : Google Scholar : PubMed/NCBI
3	Fahy RJ and Wewers MD: Pulmonary defense and the human cathelicidin hCAP-18/LL-37. Immunol Res. 31:75–89. 2005. View Article : Google Scholar : PubMed/NCBI
4	Gennaro R and Zanetti M: Structural features and biological activities of the cathelicidin-derived antimicrobial peptides. Biopolymers. 55:31–49. 2015. View Article : Google Scholar : PubMed/NCBI
5	Wang X, Mishra B, Lushnikova T, Narayana JL and Wang G: Amino acid composition determines peptide activity spectrum and hot-spot-based design of merecidin. Adv Biosyst. 2:17002592018. View Article : Google Scholar : PubMed/NCBI
6	Wang G, Hanke ML, Mishra B, Lushnikova T, Heim CE, Chittezham Thomas V, Bayles KW and Kielian T: Transformation of human cathelicidin LL-37 into selective, stable, and potent antimicrobial compounds. ACS Chem Biol. 9:1997–2002. 2014. View Article : Google Scholar : PubMed/NCBI
7	Zasloff M: Antimicrobial peptides of multicellular organisms. Nature. 415:389–395. 2002. View Article : Google Scholar : PubMed/NCBI
8	Mader JS, Mookherjee N, Hancock RE and Bleackley RC: The human host defense peptide LL-37 induces apoptosis in a calpain- and apoptosis-inducing factor-dependent manner involving Bax activity. Mol Cancer Res. 7:689–702. 2009. View Article : Google Scholar : PubMed/NCBI
9	Xu Z, Zhang F, Bai C, Yao C, Zhong H, Zou C and Chen X: Sophoridine induces apoptosis and S phase arrest via ROS-dependent JNK and ERK activation in human pancreatic cancer cells. J Exp Clin Cancer Res. 36:1242017. View Article : Google Scholar : PubMed/NCBI
10	McCubrey JA, Steelman LS, Chappell WH, Abrams SL, Wong EW, Chang F, Lehmann B, Terrian DM, Milella M, Tafuri A, et al: Roles of the Raf/MEK/ERK pathway in cell growth, malignant transformation and drug resistance. Biochim Biophys Acta. 1773:1263–1284. 2007. View Article : Google Scholar : PubMed/NCBI
11	Le Chevalier T: Adjuvant chemotherapy for resectable non-small-cell lung cancer: Where is it going? Ann Oncol. 21 (Suppl 7):vii196–vii198. 2010. View Article : Google Scholar : PubMed/NCBI
12	Hu L, Zhang T, Liu D, Guan G, Huang J, Proksch P, Chen X and Lin W: Notoamide-type alkaloid induced apoptosis and autophagy via a P38/JNK signaling pathway in hepatocellular carcinoma cells. RSC Adv. 9:19855–19868. 2019. View Article : Google Scholar
13	Tian SW, Ren Y, Pei JZ, Ren BC and He Y: Pigment epithelium-derived factor protects retinal ganglion cells from hypoxia-induced apoptosis by preventing mitochondrial dysfunction. Int J Ophthalmol. 10:1046–1054. 2017.PubMed/NCBI
14	Kang J, Dietz MJ and Li B: Antimicrobial peptide LL-37 is bactericidal against Staphylococcus aureus biofilms. PLoS One. 14:e02166762019. View Article : Google Scholar : PubMed/NCBI
15	Ko JK and Zhang Z: LL-37 inhibits pancreatic cancer development through inhibition of autophagy and reprogramming of the tumor microenvironment. Pergamon. 110:S5–S6. 2019.
16	Agerberth B, Charo J, Werr J, Olsson B, Idali F, Lindbom L, Kiessling R, Jörnvall H, Wigzell H and Gudmundsson GH: The human antimicrobial and chemotactic peptides LL-37 and alpha-defensins are expressed by specific lymphocyte and monocyte populations. Blood. 96:3086–3093. 2000. View Article : Google Scholar : PubMed/NCBI
17	Dale BA and Fredericks LP: Antimicrobial peptides in the oral environment: Expression and function in health and disease. Curr Issues Mol Biol. 7:119–133. 2005.PubMed/NCBI
18	Rico-Mata R, De Leon-Rodriguez LM and Avila EE: Effect of antimicrobial peptides derived from human cathelicidin LL-37 on Entamoeba histolytica trophozoites. Exp Parasitol. 133:300–306. 2013. View Article : Google Scholar : PubMed/NCBI
19	Rekha RS, Rao Muvva SS, Wan M, Raqib R, Bergman P, Brighenti S, Gudmundsson GH and Agerberth B: Phenylbutyrate induces LL-37-dependent autophagy and intracellular killing of Mycobacterium tuberculosis in human macrophages. Autophagy. 11:1688–1699. 2015. View Article : Google Scholar : PubMed/NCBI
20	Bucki R, Leszczyńska K, Namiot A and Sokołowski W: Cathelicidin LL-37: A multitask antimicrobial peptide. Arch Immunol Ther Exp (Warsz). 58:15–25. 2010. View Article : Google Scholar : PubMed/NCBI
21	Dosler S and Karaaslan E: Inhibition and destruction of Pseudomonas aeruginosa biofilms by antibiotics and antimicrobial peptides. Peptides. 62:32–37. 2014. View Article : Google Scholar : PubMed/NCBI
22	Amatngalim GD, Nijnik A, Hiemstra PS and Hancock RE: Cathelicidin Peptide LL-37 Modulates TREM-1 expression and inflammatory responses to microbial compounds. Inflammation. 34:412–425. 2011. View Article : Google Scholar : PubMed/NCBI
23	Shaykhiev R, Beisswenger C, Kändler K, Senske J, Püchner A, Damm T, Behr J and Bals R: Human endogenous antibiotic LL-37 stimulates airway epithelial cell proliferation and wound closure. Am J Physiol Lung Cell Mol Physiol. 289:L842–L848. 2005. View Article : Google Scholar : PubMed/NCBI
24	Pan G: Inhibitory effect of antimicrobial peptide LL-37 on bladder tumor cells (unpublished PhD thesis). Kunming Medical University; 2015
25	Ren SX, Shen J, Cheng AS, Lu L, Chan RL, Li ZJ, Wang XJ, Wong CC, Zhang L, Ng SS, et al: FK-16 Derived from the Anticancer Peptide LL-37 induces caspase-independent apoptosis and autophagic cell death in colon cancer cells. PLoS One. 8:e636412013. View Article : Google Scholar : PubMed/NCBI
26	Coffelt SB, Waterman RS, Florez L, Höner zu Bentrup K, Zwezdaryk KJ, Tomchuck SL, LaMarca HL, Danka ES, Morris CA and Scandurro AB: Ovarian cancers overexpress the antimicrobial protein hCAP-18 and its derivative LL-37 increases ovarian cancer cell proliferation and invasion. Int J Cancer. 122:1030–1039. 2008. View Article : Google Scholar : PubMed/NCBI
27	Kim JE, Kim HJ, Choi JM, Lee KH, Kim TY, Cho BK, Jung JY, Chung KY, Cho D and Park HJ: The antimicrobial peptide human cationic antimicrobial protein-18/cathelicidin LL-37 as a putative growth factor for malignant melanoma. Br J Dermatol. 163:959–967. 2010. View Article : Google Scholar : PubMed/NCBI
28	Von Haussen J, Koczulla R, Shaykhiev R, Herr C, Pinkenburg O, Reimer D, Wiewrodt R, Biesterfeld S, Aigner A, Czubayko F and Bals R: The host defence peptide LL-37/hCAP-18 is a growth factor for lung cancer cells. Lung Cancer. 59:12–23. 2008. View Article : Google Scholar : PubMed/NCBI
29	Chieosilapatham P, Ikeda S, Ogawa H and Niyonsaba F: Tissue-specific regulation of innate immune responses by human cathelicidin LL-37. Curr Pharm Des. 24:1079–1091. 2018. View Article : Google Scholar : PubMed/NCBI
30	Ou Hongyu, Zhu Haihong, Zhu Wenjun, et al: Antimicrobial peptide LL-37 induces apoptosis in AGS cells of gastric cancer by activating p53 signaling pathway. J Anhui Med Uni. 56:571–576. 2021.
31	Vandamme D, Landuyt B, Luyten W and Schoofs L: A comprehensive summary of LL-37, the factotum human cathelicidin peptide. Cell Immunol. 280:22–35. 2012. View Article : Google Scholar : PubMed/NCBI
32	Wang G: Structures of Human host defense cathelicidin LL-37 and its smallest antimicrobial peptide KR-12 in lipid micelles. J Biol Chem. 283:32637–32643. 2008. View Article : Google Scholar : PubMed/NCBI
33	Doss M, White MR, Tecle T and Hartshorn KL: Human defensins and LL-37 in mucosal immunity. J Leukoc Biol. 87:79–92. 2010. View Article : Google Scholar : PubMed/NCBI
34	An LL, Ma XT, Yang YH, Lin YM, Song YH and Wu KF: Marked reduction of LL-37/hCAP-18, an antimicrobial peptide, in patients with acute myeloid leukemia. Int J Hematol. 81:45–47. 2005. View Article : Google Scholar : PubMed/NCBI
35	Choi SY, Kim SJ, Chi BH, Kwon JK and Chang IH: Modulating the internalization of bacille calmette-guérin by cathelicidin in bladder cancer cells. Urology. 85:964.e7–964.e12. 2015. View Article : Google Scholar : PubMed/NCBI
36	Ang MK and Mok TSK: Twenty-five years of Respirology: Advances in lung cancer. Respirology. 25:26–31. 2020. View Article : Google Scholar : PubMed/NCBI
37	Xu N, Jia D, Chen W, Wang H, Liu F, Ge H, Zhu X, Song Y, Zhang X, Zhang D, et al: FoxM1 is associated with poor prognosis of non-small cell lung cancer patients through promoting tumor metastasis. PLoS One. 8:e594122013. View Article : Google Scholar : PubMed/NCBI
38	Xuan Y, Zhao S, Xiao X, Xiang L and Zheng HC: Inhibition of chaperone-mediated autophagy reduces tumor growth and metastasis and promotes drug sensitivity in colorectal cancer. Mol Med Rep. 23:3602021. View Article : Google Scholar : PubMed/NCBI
39	Wu WK, Sung JJ, To KF, Yu L, Li HT, Li ZJ, Chu KM, Yu J and Cho CH: The host defense peptide LL-37 activates the tumor-suppressing bone morphogenetic protein signaling via inhibition of proteasome in gastric cancer cells. J Cell Physiol. 223:178–186. 2010.PubMed/NCBI
40	Murray NP, Aedo S, Fuentealba C, Salazar A, Reyes E, Lopez MA and Minzer S: Circulating prostate cells and bone marrow micro-metastasis and not treatment modality determine the risk and time to biochemical failure in low risk prostate cancer. Arch Esp Urol. 72:1000–1009. 2019.(In Spanish). PubMed/NCBI
41	Sancar A, Lindsey-Boltz LA, Unsal-Kaçmaz K and Linn S: Molecular mechanisms of mammalian DNA repair and the DNA damage checkpoints. Annu Rev Biochem. 73:39–85. 2004. View Article : Google Scholar : PubMed/NCBI
42	Liu X, Li J, Huang L, Wang Y, Yang M, Tang M and Qiu T: Preparation and evaluation of MPEG-PCL polymeric nanoparticles against gastric cancer. J Wuhan Univ Technol-Mat Sci Edit. 35:1162–1168. 2021. View Article : Google Scholar
43	Tian W, Li B, Zhang X, Dang W, Wang X, Tang H, Wang L, Cao H and Chen T: Suppression of tumor invasion and migration in breast cancer cells following delivery of siRNA against Stat3 with the antimicrobial peptide PR39. Oncol Rep. 28:1362–1368. 2012. View Article : Google Scholar : PubMed/NCBI
44	Zhu M, Miao S, Zhou W, Elnesr SS, Dong X and Zou X: MAPK, AKT/FoxO3a and mTOR pathways are involved in cadmium regulating the cell cycle, proliferation and apoptosis of chicken follicular granulosa cells. Ecotoxicol Environ Saf. 214:1120912021. View Article : Google Scholar : PubMed/NCBI
45	Bartek J, Lukas C and Lukas J: Checking on DNA damage in S phase. Nat Rev Mol Cell Biol. 5:792–804. 2004. View Article : Google Scholar : PubMed/NCBI
46	Al-Ejeh F, Kumar R, Wiegmans A, Lakhani SR, Brown MP and Khanna KK: Harnessing the complexity of DNA-damage response pathways to improve cancer treatment outcomes. Oncogene. 29:6085–6098. 2010. View Article : Google Scholar : PubMed/NCBI
47	Campos A and Clemente-Blanco A: Cell cycle and DNA repair regulation in the damage response: Protein phosphatases take over the reins. Int J Mol Sci. 21:4462020. View Article : Google Scholar : PubMed/NCBI
48	Zeng Q: Expression of antimicrobial peptide LL-37 in urothelial carcinoma of the bladder. Kunming Medical University; 2015
49	Su L, Xu G, Shen J, Tuo Y, Zhang X, Jia S, Chen Z and Su X: Anticancer bioactive peptide suppresses human gastric cancer growth through modulation of apoptosis and the cell cycle. Oncol Rep. 23:3–9. 2010.PubMed/NCBI
50	Fink SL and Cookson BT: Apoptosis, pyroptosis, and necrosis: Mechanistic description of dead and dying eukaryotic cells. Infect Immun. 73:1907–1916. 2005. View Article : Google Scholar : PubMed/NCBI
51	Li Jun: Antimicrobial peptide 17BIPHE2 inhibits lung adenocarcinoma A549 cells and its mechanism. Ningxia Medical University; 2018
52	Song J, Ham J, Hong T, Song G and Lim W: Fraxetin suppresses cell proliferation and induces apoptosis through mitochondria dysfunction in human hepatocellular carcinoma cell lines Huh7 and Hep3B. Pharmaceutics. 13:1122021. View Article : Google Scholar : PubMed/NCBI
53	Chuang KC, Chen FW, Tsai MH and Shieh JJ: EGR-1 plays a protective role in AMPK inhibitor compound C-induced apoptosis through ROS-induced ERK activation in skin cancer cells. Oncol Lett. 21:3042021. View Article : Google Scholar : PubMed/NCBI
54	Schwartz GK and Shah MA: Targeting the cell cycle: A new approach to cancer therapy. J Clin Oncol. 23:9408–9421. 2006. View Article : Google Scholar : PubMed/NCBI
55	Dong Y, Yang Y, Wei Y, Gao Y, Jiang W, Wang G and Wang D: Facile synthetic nano-curcumin encapsulated Bio-fabricated nanoparticles induces ROS-mediated apoptosis and migration blocking of human lung cancer cells. Process Biochemistry. 95:91–98. 2020. View Article : Google Scholar
56	Hong Y, Sun Y, Rong X, Li D, Lu Y and Ji Y: Exosomes from adipose-derived stem cells attenuate UVB-induced apoptosis, ROS, and the Ca²⁺ level in HLEC cells. Exp Cell Res. 396:1123212020. View Article : Google Scholar : PubMed/NCBI
57	Wang CL, Liu C, Niu LL, Wang LR, Hou LH and Cao XH: Surfactin-induced apoptosis through ROS-ERS-Ca2+-ERK pathways in HepG2 cells. Cell Biochem Biophys. 67:1433–1439. 2013. View Article : Google Scholar : PubMed/NCBI
58	Kuriyama I, Miyazaki A, Tsuda Y, Yoshida H and Mizushina Y: Inhibitory effect of novel somatostatin peptide analogues on human cancer cell growth based on the selective inhibition of DNA polymerase β. Bioorg Med Chem. 21:403–411. 2013. View Article : Google Scholar : PubMed/NCBI
59	Cabrera Zapata LE, Bollo M and Cambiasso MJ: Estradiol-mediated axogenesis of hypothalamic neurons requires ERK1/2 and ryanodine receptors-dependent intracellular Ca²⁺ rise in male rats. Front Cell Neurosci. 13:1222019. View Article : Google Scholar : PubMed/NCBI
60	Adams JM and Cory S: The Bcl-2 protein family: Arbiters of cell survival. Science. 281:1322–1326. 1998. View Article : Google Scholar : PubMed/NCBI
61	Liu Y, et al: HSV-2 miR-H4-5p negatively regulates CDKL2 gene expression, blocking actinomycin D (ActD)-induced apoptosis in vero cells. Chin J Biochem Mol Bio. 17:9728–9735. 2015.PubMed/NCBI
62	Desagher S and Martinou JC: Mitochondria as the central control point of apoptosis. Trends in Cell Biol. 10:369–377. 2000. View Article : Google Scholar : PubMed/NCBI
63	Wang M, Lu X, Dong X, Hao F, Liu Z, Ni G and Chen D: pERK1/2 silencing sensitizes pancreatic cancer BXPC-3 cell to gemcitabine-induced apoptosis via regulating Bax and Bcl-2 expression. World J Surg Oncol. 13:662015. View Article : Google Scholar : PubMed/NCBI
64	Wang Z, Xu Z, Niu Z, Liang B and Niu J: Epieriocalyxin A induces cell apoptosis through JNK and ERK1/2 signaling pathways in colon cancer cells. Cell Biochem Biophys. 73:559–564. 2015. View Article : Google Scholar : PubMed/NCBI