Amino Acids

, Volume 50, Issue 5, pp 577–592 | Cite as

Selection and identification of novel peptides specifically targeting human cervical cancer

Original Article


Cervical cancer is the second most commonly diagnosed cancer and the third leading cause of cancer deaths among females in underdeveloped countries. This study aimed to identify several novel cervical cancer-specific targeting peptides (CSPs) to provide new methods for the effective diagnosis and treatment of cervical cancer. Peptide library screening in vivo was performed on human cervical cancer xenografts with Ph.D.™-12 and C7C phage display peptide libraries. Two specific peptide sequences (GDALFSVPLEVY and KQNLAEG), which were enriched in tumors, were screened, and respectively, named CSP-GD and CSP-KQ through three rounds of biopanning. The in vivo tumor-targeting ability of these peptides was identified by injecting them into mice with cervical cancer xenograft. CSPs were compounded and labeled with fluorescein isothiocyanate (FITC). The specificity and affinity of FITC-CSPs were evaluated in human cervical cancer cell lines and tissue microarrays in vitro by immunofluorescent staining. Results showed that FITC-CSP-GD and FITC-CSP-KQ evidently and specifically bound to the cell membrane and cytoplasm of SiHa, ME-180, and C-33A cells in vitro. In human cervical cancer tissue, FITC-CSP-GD and FITC-CSP-KQ strongly targeted human cervical adenocarcinoma and cervical squamous cell carcinoma tissues, respectively. A bright FITC signal was located mainly on the cell membrane and cytoplasm of tumor cells. In conclusion, the novel 12-residue peptide CSP-GD and 7-residue peptide CSP-KQ could specifically target human cervical cancer and may have the potential to be used in the diagnosis and targeted therapy of cervical cancer.


Peptides Tumor targeting Cervical cancer Phage display 



We would like to express our gratitude to Dr. Tang and Dr. Liang, who provided valuable guidance in each stage of our paper writing. We would also like to show our appreciation to all individuals who contributed to this paper. This study was supported by the National Natural Science Foundation of China (Grant no. 81472449 and 81101988), the Hunan Provincial Education Department Foundation of China (Grant no. 11C1091), and the Hengyang Industry University Research Project in Hunan Province of China (Grant no. 2015kc53). The study was performed using equipment purchased with funding from the construction program of the key discipline in Hunan Province, China (Basic Medicine Sciences in University of South China).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Bastien JI, McNeill KA, Fine HA (2015) Molecular characterizations of glioblastoma, targeted therapy, and clinical results to date. Cancer 121(4):502–516. CrossRefPubMedGoogle Scholar
  2. Brown KC (2010) Peptidic tumor targeting agents: the road from phage display peptide selections to clinical applications. Curr Pharm Des 16(9):1040–1054CrossRefPubMedPubMedCentralGoogle Scholar
  3. Chen B, Cao S, Zhang Y, Wang X, Liu J, Hui X, Wan Y, Du W, Wang L, Wu K, Fan D (2009) A novel peptide (GX1) homing to gastric cancer vasculature inhibits angiogenesis and cooperates with TNF alpha in anti-tumor therapy. BMC Cell Biol 10:63. CrossRefPubMedPubMedCentralGoogle Scholar
  4. Chen K, Sun X, Niu G, Ma Y, Yap LP, Hui X, Wu K, Fan D, Conti PS, Chen X (2012a) Evaluation of 64Cu labeled GX1: a phage display peptide probe for PET imaging of tumor vasculature. Mol Imaging Biol 14(1):96–105. CrossRefPubMedPubMedCentralGoogle Scholar
  5. Chen K, Yap LP, Park R, Hui X, Wu K, Fan D, Chen X, Conti PS (2012b) A Cy5.5-labeled phage-displayed peptide probe for near-infrared fluorescence imaging of tumor vasculature in living mice. Amino Acids 42(4):1329–1337. CrossRefPubMedGoogle Scholar
  6. Chen W, Zheng R, Baade PD, Zhang S, Zeng H, Bray F, Jemal A, Yu XQ, He J (2016) Cancer statistics in China, 2015. CA Cancer J Clin 66(2):115–132. CrossRefPubMedGoogle Scholar
  7. Corso S, Ghiso E, Cepero V, Sierra JR, Migliore C, Bertotti A, Trusolino L, Comoglio PM, Giordano S (2010) Activation of HER family members in gastric carcinoma cells mediates resistance to MET inhibition. Mol Cancer 9:121. CrossRefPubMedPubMedCentralGoogle Scholar
  8. Crafton SM, Salani R (2016) Beyond chemotherapy: an overview and review of targeted therapy in cervical cancer. Clin Ther 38(3):449–458. CrossRefPubMedGoogle Scholar
  9. Dai Y, Yin J, Huang Y, Chen X, Wang G, Liu Y, Zhang X, Nie Y, Wu K, Liang J (2016) In vivo quantifying molecular specificity of Cy5.5-labeled cyclic 9-mer peptide probe with dynamic fluorescence imaging. Biomed Opt Express 7(4):1149–1159. CrossRefPubMedPubMedCentralGoogle Scholar
  10. de Freitas AC, Gomes Leitao Mda C, Coimbra EC (2015) Prospects of molecularly-targeted therapies for cervical cancer treatment. Curr Drug Targets 16(1):77–91CrossRefPubMedGoogle Scholar
  11. de Melo AC, Paulino E, Garces AH (2017) A Review of mTOR pathway inhibitors in gynecologic cancer. Oxid Med Cell Longev 2017:4809751. CrossRefPubMedPubMedCentralGoogle Scholar
  12. Du Y, Zhang Q, Jing L, Liang X, Chi C, Li Y, Yang X, Dai Z, Tian J (2015) GX1-conjugated poly(lactic acid) nanoparticles encapsulating Endostar for improved in vivo anticolorectal cancer treatment. Int J Nanomed 10:3791–3802. CrossRefGoogle Scholar
  13. Ferlay J, Soerjomataram I, Ervik M, Dikshit R, Eser S, Mathers C, Rebelo M, Parkin D, Forman D, Bray F (2013) GLOBOCAN 2012 v1.0, Cancer Incidence and Mortality Worldwide: IARC CancerBase No. 11. Lyon, France: International Agency for Research on Cancer.
  14. Fetcko K, Gondim DD, Bonnin JM, Dey M (2017) Cervical cancer metastasis to the brain: a case report and review of literature. Surg Neurol Int 8:181. CrossRefPubMedPubMedCentralGoogle Scholar
  15. Fu HC, Yang YC, Chen YJ, Lin H, Ou YC, Chien CC, Huang EY, Huang HY, Lan J, Chi HP, Huang KE, Kang HY (2016) Increased expression of SKP2 is an independent predictor of locoregional recurrence in cervical cancer via promoting DNA-damage response after irradiation. Oncotarget 7(28):44047–44061. CrossRefPubMedPubMedCentralGoogle Scholar
  16. Geiss R, De La Motte Rouge T, Dubot C, Leary A, Lhomme C, Pautier P, Scholl S, Rodrigues MJ (2014) Targeted therapy in locally and metastatic recurrent cervical cancers. Bull Cancer 101(7–8):748–755. PubMedGoogle Scholar
  17. Guan M, Wang J, Yang L, Zhao Z, Lu K, Zhao L, Xiao J, Li Z, Shi Z (2014) Targeting osteosarcoma vasculature with peptide obtained by phage display. Contemp Oncol 18(3):165–170. Google Scholar
  18. Hagimori M, Fuchigami Y, Kawakami S (2017) Peptide-Based cancer-targeted DDS and molecular imaging. Chem Pharm Bull 65(7):618–624. CrossRefPubMedGoogle Scholar
  19. Henry KA, Arbabi-Ghahroudi M, Scott JK (2015) Beyond phage display: non-traditional applications of the filamentous bacteriophage as a vaccine carrier, therapeutic biologic, and bioconjugation scaffold. Front Microbiol 6:755. PubMedPubMedCentralGoogle Scholar
  20. Herold DM, Das IJ, Stobbe CC, Iyer RV, Chapman JD (2000) Gold microspheres: a selective technique for producing biologically effective dose enhancement. Int J Radiat Biol 76(10):1357–1364CrossRefPubMedGoogle Scholar
  21. Kaplan G, Gershoni JM (2012) A general insert label for peptide display on chimeric filamentous bacteriophages. Anal Biochem 420(1):68–72. CrossRefPubMedGoogle Scholar
  22. Kim HS, Kim T, Lee ES, Kim HJ, Chung HH, Kim JW, Song YS, Park NH (2013) Impact of chemoradiation on prognosis in stage IVB cervical cancer with distant lymphatic metastasis. Cancer Res Treat 45(3):193–201. CrossRefPubMedPubMedCentralGoogle Scholar
  23. Lee KJ, Lee JH, Chung HK, Choi J, Park J, Park SS, Ju EJ, Park J, Shin SH, Park HJ, Ko EJ, Suh N, Kim I, Hwang JJ, Song SY, Jeong SY, Choi EK (2015) Novel peptides functionally targeting in vivo human lung cancer discovered by in vivo peptide displayed phage screening. Amino Acids 47(2):281–289. CrossRefPubMedGoogle Scholar
  24. Lee KJ, Lee JH, Chung HK, Ju EJ, Song SY, Jeong SY, Choi EK (2016) Application of peptide displaying phage as a novel diagnostic probe for human lung adenocarcinoma. Amino Acids 48(4):1079–1086. CrossRefPubMedGoogle Scholar
  25. Li M, Anastassiades CP, Joshi B, Komarck CM, Piraka C, Elmunzer BJ, Turgeon DK, Johnson TD, Appelman H, Beer DG, Wang TD (2010) Affinity peptide for targeted detection of dysplasia in Barrett’s esophagus. Gastroenterology 139(5):1472–1480. CrossRefPubMedPubMedCentralGoogle Scholar
  26. Li J, Feng L, Fan L, Zha Y, Guo L, Zhang Q, Chen J, Pang Z, Wang Y, Jiang X, Yang VC, Wen L (2011) Targeting the brain with PEG-PLGA nanoparticles modified with phage-displayed peptides. Biomaterials 32(21):4943–4950. CrossRefPubMedPubMedCentralGoogle Scholar
  27. Li J, Zhang Q, Pang Z, Wang Y, Liu Q, Guo L, Jiang X (2012) Identification of peptide sequences that target to the brain using in vivo phage display. Amino Acids 42(6):2373–2381. CrossRefPubMedGoogle Scholar
  28. Li H, Wu X, Cheng X (2016) Advances in diagnosis and treatment of metastatic cervical cancer. J Gynecol Oncol 27(4):e43. CrossRefPubMedPubMedCentralGoogle Scholar
  29. Li C, Gao N, Xue Q, Ma N, Hu Y, Zhang J, Chen B, Hou Y (2017) Screening and identification of a specific peptide binding to cervical cancer cells from a phage-displayed peptide library. Biotech Lett 39(10):1463–1469. CrossRefGoogle Scholar
  30. Liu X, Li S, Yi F (2014) Trop2 gene: a novel target for cervical cancer treatment. J Cancer Res Clin Oncol 140(8):1331–1341. CrossRefPubMedGoogle Scholar
  31. Ma C, Yin G, Yan D, He X, Zhang L, Wei Y, Huang Z (2013) A novel peptide specifically targeting ovarian cancer identified by in vivo phage display. J Pept Sci 19(12):730–736. CrossRefPubMedGoogle Scholar
  32. Mandelin J, Cardo-Vila M, Driessen WH, Mathew P, Navone NM, Lin SH, Logothetis CJ, Rietz AC, Dobroff AS, Proneth B, Sidman RL, Pasqualini R, Arap W (2015) Selection and identification of ligand peptides targeting a model of castrate-resistant osteogenic prostate cancer and their receptors. Proc Natl Acad Sci USA 112(12):3776–3781. PubMedPubMedCentralGoogle Scholar
  33. Martinho O, Silva-Oliveira R, Cury FP, Barbosa AM, Granja S, Evangelista AF, Marques F, Miranda-Goncalves V, Cardoso-Carneiro D, de Paula FE, Zanon M, Scapulatempo-Neto C, Moreira MA, Baltazar F, Longatto-Filho A, Reis RM (2017) HER family receptors are important theranostic biomarkers for cervical cancer: blocking glucose metabolism enhances the therapeutic effect of her inhibitors. Theranostics 7(3):717–732. CrossRefPubMedPubMedCentralGoogle Scholar
  34. Masoumi Moghaddam S, Amini A, Morris DL, Pourgholami MH (2012) Significance of vascular endothelial growth factor in growth and peritoneal dissemination of ovarian cancer. Cancer Metastasis Rev 31(1–2):143–162. CrossRefPubMedGoogle Scholar
  35. Newton J, Deutscher SL (2008) Phage peptide display. Handbook Exp Pharmacol 185:145–163. CrossRefGoogle Scholar
  36. Newton JR, Deutscher SL (2009) In vivo bacteriophage display for the discovery of novel peptide-based tumor-targeting agents. Methods Mol Biol 504:275–290. CrossRefPubMedGoogle Scholar
  37. NIH (2017) SEER data: surveillance, epidemiology and end results. National Cancer Institute.
  38. Noren KA, Noren CJ (2001) Construction of high-complexity combinatorial phage display peptide libraries. Methods 23(2):169–178. CrossRefPubMedGoogle Scholar
  39. Oh DY, Kim S, Choi YL, Cho YJ, Oh E, Choi JJ, Jung K, Song JY, Ahn SE, Kim BG, Bae DS, Park WY, Lee JW, Song S (2015) HER2 as a novel therapeutic target for cervical cancer. Oncotarget 6(34):36219–36230. CrossRefPubMedPubMedCentralGoogle Scholar
  40. Pasqualini R, Ruoslahti E (1996) Organ targeting in vivo using phage display peptide libraries. Nature 380(6572):364–366. CrossRefPubMedGoogle Scholar
  41. Robinson P, Stuber D, Deryckere F, Tedbury P, Lagrange M, Orfanoudakis G (2005) Identification using phage display of peptides promoting targeting and internalization into HPV-transformed cell lines. J Mol Recognit 18(2):175–182. CrossRefPubMedGoogle Scholar
  42. Seol HJ, Ulak R, Ki KD, Lee JM (2014) Cytotoxic and targeted systemic therapy in advanced and recurrent cervical cancer: experience from clinical trials. Tohoku J Exp Med 232(4):269–276CrossRefPubMedGoogle Scholar
  43. Silacci M, Brack S, Schirru G, Marlind J, Ettorre A, Merlo A, Viti F, Neri D (2005) Design, construction, and characterization of a large synthetic human antibody phage display library. Proteomics 5(9):2340–2350. CrossRefPubMedGoogle Scholar
  44. Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J, Jemal A (2015) Global cancer statistics, 2012. CA Cancer J Clin 65(2):87–108. CrossRefPubMedGoogle Scholar
  45. Trepel M, Arap W, Pasqualini R (2002) In vivo phage display and vascular heterogeneity: implications for targeted medicine. Curr Opin Chem Biol 6(3):399–404CrossRefPubMedGoogle Scholar
  46. van Meir H, Kenter GG, Burggraaf J, Kroep JR, Welters MJ, Melief CJ, van der Burg SH, van Poelgeest MI (2014) The need for improvement of the treatment of advanced and metastatic cervical cancer, the rationale for combined chemo-immunotherapy. Anticancer Agents Med Chem 14(2):190–203CrossRefPubMedGoogle Scholar
  47. Velasco-Velazquez M, Xolalpa W, Pestell RG (2014) The potential to target CCL5/CCR5 in breast cancer. Expert Opin Ther Targets 18(11):1265–1275. CrossRefPubMedGoogle Scholar
  48. Wolfe T, Chatterjee D, Lee J, Grant JD, Bhattarai S, Tailor R, Goodrich G, Nicolucci P, Krishnan S (2015) Targeted gold nanoparticles enhance sensitization of prostate tumors to megavoltage radiation therapy in vivo. Nanomed Nanotechnol Biol Med 11(5):1277–1283. CrossRefGoogle Scholar
  49. Xu G, Yao Z, Yang H, Liu Y (2015) The effect of gold nanoparticles on increasing radiosensitivity of SPC-A1 lung cancer cell in vitro. Chinese Clin Oncol 20(6):492–496Google Scholar
  50. Yang P (2010) Panning peptides targeting to hepatocellular carcinoma and the targeting study of X1/MePEG-PLA-CS nanoparticles. Dissertation, Central South UniversityGoogle Scholar
  51. Yang W, Luo D, Wang S, Wang R, Chen R, Liu Y, Zhu T, Ma X, Liu R, Xu G, Meng L, Lu Y, Zhou J, Ma D (2008) TMTP1, a novel tumor-homing peptide specifically targeting metastasis. Clinical Cancer Res 14(17):5494–5502. CrossRefGoogle Scholar
  52. Yang C, He X, Liu X, Tang Z, Liang X (2015) OSTP as a novel peptide specifically targeting human ovarian cancer. Oncol Rep 34(2):972–978. CrossRefPubMedGoogle Scholar
  53. Yang X, Zhang F, Luo J, Pang J, Yan S, Luo F, Liu J, Wang W, Cui Y, Su X (2016) A new non-muscle-invasive bladder tumor-homing peptide identified by phage display in vivo. Oncol Rep 36(1):79–89. CrossRefPubMedPubMedCentralGoogle Scholar
  54. Zhi M, Wu KC, Dong L, Hao ZM, Deng TZ, Hong L, Liang SH, Zhao PT, Qiao TD, Wang Y, Xu X, Fan DM (2004) Characterization of a specific phage-displayed Peptide binding to vasculature of human gastric cancer. Cancer Biol Ther 3(12):1232–1235CrossRefPubMedGoogle Scholar
  55. Zhu AX, Hezel AF (2011) Development of molecularly targeted therapies in biliary tract cancers: reassessing the challenges and opportunities. Hepatology 53(2):695–704. CrossRefPubMedGoogle Scholar
  56. Zwick MB, Shen J, Scott JK (1998) Phage-displayed peptide libraries. Curr Opin Biotechnol 9(4):427–436CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Hunan Province Key Laboratory of Tumor Cellular & Molecular Pathology, Cancer Research InstituteUniversity of South ChinaHengyangPeople’s Republic of China
  2. 2.Hunan Province Cooperative Innovation Center for Molecular Target New Drug StudyUniversity of South ChinaHengyangPeople’s Republic of China
  3. 3.Department of Gynecology and Obstetrics, The Second Affiliated HospitalUniversity of South ChinaHengyangPeople’s Republic of China
  4. 4.Institute of Pharmacy and PharmacologyUniversity of South ChinaHengyangPeople’s Republic of China
  5. 5.Biomedical Research CenterHunan University of MedicineHuaihuaPeople’s Republic of China

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