Molecular design and anti‑melanoma activity of a novel bullfrog antibacterial peptide RGD‑chimera

Liu,Mengyue; Jiang,Xuan; Fu,Chao; Zhao,Ruili; Jin,Tianming; Ma,Jifei; Qin,Shunyi; Li,Liu An; Hu,Ye; Zhang,Xin

doi:10.3892/ol.2020.12376

Molecular design and anti‑melanoma activity of a novel bullfrog antibacterial peptide RGD‑chimera

Authors:
- Mengyue Liu
- Xuan Jiang
- Chao Fu
- Ruili Zhao
- Tianming Jin
- Jifei Ma
- Shunyi Qin
- Liu An Li
- Ye Hu
- Xin Zhang
View Affiliations

Affiliations: Tianjin Key Laboratory of Agricultural Animal Breeding and Healthy Husbandry, College of Animal Science and Veterinary Medicine, Tianjin Agricultural University, Tianjin 300384, P.R. China
Published online on: December 15, 2020 https://doi.org/10.3892/ol.2020.12376
Article Number: 115
Copyright: © Liu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Melanoma is a common malignant skin tumor, which is the only fatal skin tumor at present. Melanoma has a high degree of malignancy and metastasis. The activity of modified Temporin‑La (T‑La) peptides from bullfrog skin were evaluated for antitumor activity and improved targeting in melanoma cells. The amino acid sequence of T‑La was modified, resulting in the antitumor peptide, T‑La (FS). T‑La and T‑La (FS) were coupled to the RGD small molecule polypeptide to form the chimeric peptides RGD‑T‑La and RGD‑T‑La (FS), respectively. The secondary structures for the peptides, evaluated using circular dichroism, were found to be α‑helical. The structure of T‑La was evaluated using bioinformatics. In addition, the antitumor effects of the modified peptide and the targeting of RGD chimeric peptide to the tumor in vivo and in vitro were analyzed. Antitumor activity was measured in vitro using the MTT assay. Tumor cells with high integrin αvβ3 expression were detected using flow cytometry, and tumor cells were screened for sensitivity to RGD‑T‑La (FS) to establish a tumor model in nude mice. The effects of the peptides on tumor cells were measured using laser confocal microscopy in real‑time. The mechanism of the peptide antitumor activity in tumor cells was evaluated with scanning electron microscopy. B16 melanoma cells were the most sensitive to the peptides, for which the cell survival rate was 24.65% for 10 µg/ml RGD‑T‑La (FS). RGD‑La (FS) had a rapid effect on tumor cells. RGD chimeric polypeptides exhibited site‑targeting cytotoxic effects in tumor cells. In the B16 melanoma mouse model, the peptides exhibited antitumor effects against early melanoma development and induced tumor apoptosis, possibly by inhibiting VEGF and promoting caspase‑3 expression. Overall, the present study provides a scientific basis for the application of small molecule antimicrobial peptides as targeted antitumor agents and lays the foundation for the clinical application of these peptides as antitumor drugs.

View Figures

View References

1	Li S, Hao L, Bao W, Zhang P, Su D, Cheng Y, Nie L, Wang G, Hou F and Yang Y: A novel short anionic antibacterial peptide isolated from the skin of Xenopus laevis with broad antibacterial activity and inhibitory activity against breast cancer cell. Arch Microbiol. 198:473–482. 2016. View Article : Google Scholar : PubMed/NCBI
2	Zhang F and Li MS: Antibacterial peptides' roles in immune defense and anti-tumorogenesis. World Not Antibiot. 38:284–289. 2017.(In Chinese).
3	Gao J, Xie C and Lu WY: Progress in research of antitumor activity of antimicrobial peptides. World Clinical Drugs,. 10:122012.
4	Zhu LZR, Liu SS, Ma JF, Jin TM, Liu JF, Hu Y and Liu MY: Biological activity and in vivo tissue targeting distribution of catesbeianalctin. Chin J Vet Sci. 36:185–190. 2016.(In Chinese).
5	Diao Y, Han W, Zhao H, Zhu S, Liu X, Feng X, Gu J, Yao C, Liu S, Sun C, et al: Designed synthetic analogs of the α-helical peptide temporin-La with improved antitumor efficacies via charge modification and incorporation of the integrin αvβ3 homing domain. J Pept Sci. 18:476–486. 2012. View Article : Google Scholar : PubMed/NCBI
6	Ma L, Cai ZL, Wang QR and Lei CX: Study on construction of citrostatin and its bioactivity. Chin J Biochem Pharm. 31:19–22. 2010.
7	Huang R, Long J and Zhang Y: Synthesise and activity of dual targeted antineoplastic polypeptide RGDSY-CTTHWGFTLC. Cancer Res Prev Treat. 41:531–535. 2014.
8	Ma Q, Dong N, Cao Y and Shan A: Rational design of alpha-helical antimicrobial peptide with Val and Arg residues. Acta Microbiol Sin. 51:346–451. 2011.
9	Wang XX, Niu B, Chen XJ, Xie J and Zhang YH: Synthesis and activities of RGD peptide analog. J Chin Pharm Sci. 43:462–464. 2008.(In Chinese).
10	Liu KY, Li QW and Liu GY: Study progress of malignant tumor imaging of integrin αvβ3 expression with radiolabelled RGD peptides. Medical Recapitulate. 12:757–759. 2006.(In Chinese).
11	Zhang CL, Yang M and Wang RF: The structure-activity relationship of RGD peptides binding to αvβ3 integrin and radiolabeled ligand design. J Oncol. 15:76–81. 2009.
12	Yu YP, Wang Q, Liu YC and Xie Y: Molecular basis for the targeted binding of RGD-containing peptide to integrin αvβ3. Biomaterials. 35:1667–1675. 2014. View Article : Google Scholar : PubMed/NCBI
13	Ji QZ and Zhang KW: Observation of DFRKN-1 isolate by scanning electron microscope. Anhui Agric Sci. 39:33–34. 2011.
14	Seo GY, Ha Y, Park AH, Kwon OW and Kim YJ: Leathesia difformis extract inhibits α-MSH-induced melanogenesis in B16F10 cells via down-regulation of CREB signaling pathway. Int J Mol Sci. 20:E5362019. View Article : Google Scholar : PubMed/NCBI
15	Zhang CL, Wang RF and L Z: Design of ~(131)I labeling of a disulfide bridged RGD-peptide targeted to integrin αvβ3 receptor and its biodistribution and imaging in tumor bearing mice. J Oncol. 16:276–280. 2010.
16	Dia YW: Design for tumor-targeting antimicrobial peptide chimera and its anticancer mechanism study (unpublished PhD thesis). Jilin University; 2012
17	Li ZDLY and Zhen YS: Potentiation of boanmycin antitumor activity by chemotactic peptide. Chin J Cancer Res. 17:79–83. 2005.(In Chinese). View Article : Google Scholar
18	Zhao RL, Han JY, Han WY, Liu SS, Ma JF, Lei LC, Feng X, Zhang AG and Wang X: The antibacterial and antitumor function of a novel bio-active peptide Catesbeianin-1a of Rana catesbeiana. Chin J Vet Sci. 33:1407–1411. 2013.(In Chinese).
19	Wang LZ and Min Y: Advance of” 1” 8F labeled RGD peptide as integrin α_Vβ_3 receptor imaging agents. J Isot. 24:68–75. 2011.
20	Jiang Z, Vasil AI, Hale JD, Hancock RE, Vasil ML and Hodges RS: Effects of net charge and the number of positively charged residues on the biological activity of amphipathic α-helical cationic antimicrobial peptides. Biopolymers. 90:369–383. 2008. View Article : Google Scholar : PubMed/NCBI
21	Zhang J, Xiao X, Zhu J, Gao Z, Lai X, Zhu X and Mao G: Lactoferrin- and RGD-comodified, temozolomide and vincristine-coloaded nanostructured lipid carriers for gliomatosis cerebri combination therapy. Int J Nanomedicine. 13:3039–3051. 2018. View Article : Google Scholar : PubMed/NCBI
22	Chen T, Chen L, He H, Peng J, Guo X, Zheng Y and Shao C: Influence of HAnps carrier system with doping Mg and grafting RGD on endocytosis of MG63 cells. J Cent South Univ. 48:625–634. 2017.
23	Habibi N, Kamaly N, Memic A and Shafiee H: Self-assembled peptide-based nanostructures: Smart nanomaterials toward targeted drug delivery. Nano Today. 11:41–60. 2016. View Article : Google Scholar : PubMed/NCBI
24	Lambert E, Fuselier E, Ramont L, Brassart B, Dukic S, Oudart JB, Dupont-Deshorgue A, Sellier C, Machado C and Dauchez M: Conformation-dependent binding of a Tetrastatin peptide to αvβ3 integrin decreases melanoma progression through FAK/PI₃ K/Akt pathway inhibition. Sci Rep. 8:98372018. View Article : Google Scholar : PubMed/NCBI
25	Hou J, Diao Y, Li W, Yang Z, Zhang L, Chen Z and Wu Y: RGD peptide conjugation results in enhanced antitumor activity of PD0325901 against glioblastoma by both tumor-targeting delivery and combination therapy. Int J Pharm. 505:329–340. 2016. View Article : Google Scholar : PubMed/NCBI
26	Chen G, Yu F, Sheng G, Wang J and Qu B: Progress and future direction of vascular mimicry. J Mod Oncol. 1:138–141. 2017.
27	Cao QZ and Lin ZB: Antitumor and anti-angiogenic activity of Ganoderma lucidum polysaccharides peptide. Acta Pharmacol Sin. 25:833–838. 2004.PubMed/NCBI
28	Ren HD, Zhang L, Zhao Y, Si XH, Zhang L and Wang C: Study on inhibitory effect of antimicrobial peptide Temporin-1CEa and its analogues inhibit metastasis and invasion of human melanoma A375 cells and reduce angiogenesis. Chin J Biochem Pharm. 2:19–22. 2016.(In Chinese).