A novel anti‑adhesion peptide (β3) inhibits hepatocellular carcinoma activity in vitro and in vivo

Wang,Songmei; Zhu,Jun; Liu,Yinkun

doi:10.3892/ol.2016.5277

A novel anti‑adhesion peptide (β3) inhibits hepatocellular carcinoma activity in vitro and in vivo

Authors:
- Songmei Wang
- Jun Zhu
- Yinkun Liu
View Affiliations

Affiliations: Laboratory of Molecular Biology, Experimental Teaching Center of Basic Medicine, Fudan University, Shanghai 200032, P.R. China, Department of Pharmacy, Shanghai Chest Hospital, Shanghai 200031, P.R. China, Liver Cancer Institute, Fudan University, Shanghai 200032, P.R. China
Published online on: October 18, 2016 https://doi.org/10.3892/ol.2016.5277
Pages: 4744-4748

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

The present study aimed to investigate the blocking of tumor cell adhesion to the extracellular matrix, the prevention of tumor metastasis by the β peptide trimer β3, as well as the influence of β3 on the recurrence and survival time of hepatocellular carcinoma (HCC) nude mice model LCI‑D20 after early resection. To this end, the DNA fragment of the β3 peptide (DLYYLMDLSYSMKGGDLYYLMDLSYSMKGGDLYYLMDLSYSMK) was cloned into the expression vector pET‑His and the fusion protein His‑β3 was expressed in E. coli BL21 (DE3) plysS. The anti‑adhesion effect of β3 on the highly metastatic HCC cell line HCCLM6 to fibronectin (FN) was measured by MTT assay. The inhibition of HCCLM6 cell invasion by β3 was analyzed using a Transwell (modified Boyden chamber) system and Matrigel. The influence of β3 on the recurrence of HCC and mouse survival time after early resection was investigated using the HCC metastasis nude mice model LCI‑D20. HCCLM6 cells incubated with 10, 20, 50 or 100 µmol/l β3 for 3 h demonstrated a marked reduction in adhesion to FN. The adhesion inhibition rates were 11.8, 21.7, 37.5 and 66.4%, respectively. In addition, cell invasion was reduced by 51.3% in HCCLM6 cells cultured with 100 µmol/l β3. Treatment with β3 also inhibited tumor recurrence at the incisal edge and prolonged the survival time of LCI‑D20 mice following early resection. The present study provided evidence that β3 peptide specifically blocked the adhesion and invasion of HCCLM6 cells, inhibited HCC recurrence in vivo and prolonged the survival time of HCC nude mice LCI‑D20 following hepatectomy. Therefore, β3 may be further investigated as a novel anti‑tumor drug.

View Figures

View References

1	Fang WQ, Li SP, Zhang CQ, Xu L, Shi M, Chen MS and Li JQ: Prophylaxis and clinical treatment for surgical margin recurrence of small primary hepatocellular carcinoma. Ai Zheng. 24:834–836. 2005.PubMed/NCBI
2	Zhi X, Lin L, Yang S, Bhuvaneshwar K, Wang H, Gusev Y, Lee MH, Kallakury B, Shivapurkar N, Cahn K, et al: βII-spectrin (SPTBN1) suppresses progression of hepatocellular carcinoma and Wnt signaling by regulation of Wnt inhibitor kallistatin. Hepatology. 61:598–612. 2015. View Article : Google Scholar : PubMed/NCBI
3	Wyke JA: Overview-burgeoning promise in metastasis research. Eur J Cancer. 36:1589–1594. 2000. View Article : Google Scholar : PubMed/NCBI
4	Liotta LA, Steeg PS and Stetler-Stevenson WG: Cancer metastasis and angiogenesis: An imbalance of positive and negative regulation. Cell. 64:327–336. 1991. View Article : Google Scholar : PubMed/NCBI
5	Chu XY and Chen LB: Cellular adhesive molecular and the invasion and metastasis of neoplasm. Yixue Yanjiusheng Xuebao. 13:42–45. 2000.
6	Li FH: The inhibitory effect of bioactive peptides on neoplasm metastasis. Kouqiang Hemian Waike Zazhi. 9:231–234. 1999.
7	Liu LY, Chen ZY and Zhao TH: Investigations of a peptide with RGD and YIGSR fragments: Synthesis and its anti-tumor invasion activities. Zhongguo Xinyao Zazhi. 14:729–731. 2005.
8	Saiki I, Yoneda J, Kobayashi H, Igarashi Y, Komazawa H, Ishizaki Y, Kato I and Azuma I: Antimetastatic effect by anti-adhesion therapy with cell-adhesive peptide of fibronectin in combination with anticancer drugs. Jpn J Cancer Res. 84:326–335. 1993. View Article : Google Scholar : PubMed/NCBI
9	Liu LY, Chen ZY and Zhao TH: Synthesis of RGD identical-fork-peptide derivative with inhibitive effecton adhesiveness of advanced metastatic tumor cells. Zhongguo Xinyao Zazhi. 15:1661–1663. 2006.
10	Zhang HQ, Shinohara H, Gu N, Sasaki H and Sisido M: Cell adhesion inhibition by RGD peptides linked with a photoisomerizable nonnatural amino acid. J Southeast Univ. 17:22–26. 2001.
11	Zhao M, Wang C, Jiang X and Pen S: Synthesis of RGD containing peptides and their bioactivities. Prep Biochem Biotechnol. 32:363–380. 2002. View Article : Google Scholar : PubMed/NCBI
12	Cao K, Zhao TH, Chen ZY, Gao W, Yang HS and Shi B: The invasive capacity of human lung great cellular xancerous PG cells on reformed basement membrane and inhibition of synthetic peptides. Zhongliu Fangzhi Yanjiu. 29:20–22. 2002.
13	Okrój M, Dobrzańska-Paprocka Z, Rolka K and Bigda J: In vitro and in vivo analyses of the biological activity of RGD peptides towards Ab Bomirski melanoma. Cell Mol Biol Lett. 8:873–884. 2003.PubMed/NCBI
14	Liu J, Guo SX and Tang JG: Research progress of RGD-peptide for cancer therapy. Guowai Yixue (Zhongliuxue Fence). 30:193–197. 2003.
15	Liu YK, Nemoto A, Feng Y and Uemura T: The binding ability to matrix proteins and the inhibitory effects on cell adhesion of synthetic peptides derived from a conserved sequence of integrins. J Biochem. 121:961–968. 1997. View Article : Google Scholar : PubMed/NCBI
16	Uemura T, Nemoto A and Liu YK: Synthetic peptide derived from a conserved sequence of integrin β subunit. Res Adv in Biosci & Bioeng. 23:65–83. 2000.
17	Sun JJ, Zhou XD, Liu YK, Tang ZY, Sun RX, Zhao Y and Uemura T: Inhibitory effects of synthetic beta peptide on invasion and metastasis of liver cancer. J Cancer Res Clin Oncol. 126:595–600. 2000. View Article : Google Scholar : PubMed/NCBI
18	Wang SM, Zhu J, Li Y, Pan LF, Zha XL and Liu YK: Molecular cloning and expression of anti-tumor adhesion peptide (beta3). Sheng Wu Gong Cheng Xue Bao. 21:558–562. 2005.(In Chinese). PubMed/NCBI
19	Knutson JR, Lida J, Fields GB and McCarthy JB: CD44/chondroitin sulfate proteoglycan and alpha 2beta 1 integrin mediate human melanoma cell migration on type IV collagen and invasion of basement membranes. Mol Biol Cell. 7:383–396. 1996. View Article : Google Scholar : PubMed/NCBI
20	Sun FX, Tang ZY, Lui KD, Ye SL, Xue Q, Gao DM and Ma ZC: Establishment of a metastatic model of human hepatocellular carcinoma in nude mice via orthotopic implantation of histologically intact tissues. Int J Cancer. 66:239–243. 1996. View Article : Google Scholar : PubMed/NCBI
21	Syrigos KN and Karayiannakis AJ: Adhesion molecules as targets for the treatment of neoplastic diseases. Curr Pharm Des. 12:2849–2861. 2006. View Article : Google Scholar : PubMed/NCBI
22	Jiang CG and Xu HM: Research and application of anti-adhesion therapy in cancer metastasis. Guowai Yixue (Zhongliuxue Fence). 32:31–34. 2005.
23	Wang YH, Liu YK, Li WC, Ye SL and Tang ZY: Inhibitory effect of anti-adhesion peptides on invasion/metastasis ability of hepatocellular carcinoma cells. Zhonghua Shiyan Waike Zazhi. 21:1168–1169. 2004.
24	Liu YK, Wu WZ, Wu X, Jiang Y and Zhou XD: Liver cancer metastasis and signal transduction. In: Tang ZY. Metastasis and recurrence of hepatocellular carcinoma-basic and clinical studies. Shanghai Shanghai Scientific and technological education public house. 93–104. 2003.
25	Maeda M, Izuno Y, Kawasaki K, Kaneda Y, Mu Y, Tsutsumi Y, Nakagawa S and Mayumi T: Amino acids and peptides. XXXI. Preparation of analogs of the laminin-related peptide YIGSR and their inhibitory effect on experimental metastasis. Chem Pharm Bull (Tokyo). 46:347–350. 1998. View Article : Google Scholar : PubMed/NCBI
26	Kaneda Y, Yamamoto Y, Okada N, Tsutsuml Y, Nakagawa S, Kakiuch M, Maeda M, Kawasaki K and Mayumi T: Antimetastatic effect of synthetic Glu-Ile-Leu-Asp-Val peptide derivatives containing D-amino acids. Anticancer Drugs. 8:702–707. 1997. View Article : Google Scholar : PubMed/NCBI
27	Feng ZH, Huang B, Zhang GM, Li D and Wang HT: Inducement of antitumor-immunity by DC activated by Hsp70-H22 tumor antigen peptide. Chinese Journal of Cancer Research. 15:79–85. 2003. View Article : Google Scholar
28	Wang SM, Zhu J, Li Y, Pan LF, Zha XL and Liu YK: Inhibitory effects of β peptide and polymeric β peptide on adhesion of hepatocellular carcinoma cells to fibronectin. Shiyong Zhongliu Zazhi. 21:223–226. 2005.
29	Li NF, Gemenetzidis E, Marshall FJ, Davies D, Yu Y, Frese K, Froeling FE, Woolf AK, Feakins RM, Naito Y, et al: RhoC interacts with integrin α5β1 and enhances its trafficking in migrating pancreatic carcinoma cells. PLoS One. 8:e815752013. View Article : Google Scholar : PubMed/NCBI
30	Sengupta N and MacDonald TT: The role of matrix metalloproteinases in stromal/epithelial interactions in the gut. Physiology (Bethesda). 22:401–409. 2007. View Article : Google Scholar : PubMed/NCBI
31	Benton G, Arnaoutova I, George J, Kleinman HK and Koblinski J: Matrigel: From discovery and ECM mimicry to assays and models for cancer research. Adv Drug Deliv Rev 79–80. 3–18. 2014. View Article : Google Scholar