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. 2013 Dec 2;11(12):4773-87.
doi: 10.3390/md11124773.

A new in vitro anti-tumor polypeptide isolated from Arca inflata

Jian Xu  1 , Zhiyan Chen, Liyan Song, Lili Chen, Jianhua Zhu, Shuangshuang Lv, Rongmin Yu
Affiliations

A new in vitro anti-tumor polypeptide isolated from Arca inflata

Jian Xu et al. Mar Drugs. .

Abstract

A new in vitro anti-tumor polypeptide, coded as J2-C3, was isolated from Arca inflata Reeve and purified by diethyl-aminoethanol (DEAE)-sepharose Fast Flow anion exchange and phenyl sepharose CL-4B hydrophobic chromatography. J2-C3 was identified to be a homogeneous compound by native polyacrylamide gel electrophoresis (Native-PAGE). The purity of J2-C3 was over 99% in reversed phase-high performance liquid chromatography (RP-HPLC). The molecular weight was determined as 20,538.0 Da by electrospray-ionization mass spectrometry (ESI-MS/MS). J2-C3 was rich in Glx (Gln + Glu), Lys, and Asx (Asp + Asn) according to amino acid analysis. Four partial amino acid sequences of this peptide were determined as L/ISMEDVEESR, KNGMHSI/LDVNHDGR, AMKI/LI/LNPKKGI/LVPR and AMGAHKPPKGNEL/IGHR via MALDI-TOF/TOF-MS and de novo sequencing. Secondary structural analysis by CD spectroscopy revealed that J2-C3 had the α-helix (45.2%), β-sheet (2.9%), β-turn (26.0%) and random coil (25.9%). The anti-tumor effect of J2-C3 against human tumor cells was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and the IC₅₀ values of J2-C3 were 65.57, 93.33 and 122.95 µg/mL against A549, HT-29 and HepG2 cell lines, respectively. Therefore, J2-C3 might be developed as a potential anti-tumor agent.

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Figures

Figure 1
Figure 1
Purification of the crude proteins of A. inflata by anion exchange chromatography on a DEAE-Sepharose Fast Flow column.
Figure 2
Figure 2
Purification of fraction J2 by phenyl sepharose CL-4B hydrophobic chromatography.
Figure 3
Figure 3
Electrophoresis. Purity and molecular weight analysis of J2-C3 by SDS-PAGE: (a) Lane 1, molecular weight marker; Lane 2, J2-C3. (b) Native-PAGE of J2-C3: Lane 1, J2-C3. (c) Isoelectric point determination of J2-C3 by IEF-PAGE: Lane 1, J2-C3.
Figure 4
Figure 4
RP-HPLC profile of J2-C3. Performed on an Agilent 1100 HPLC system fitted with a ZORBAX®300SB-C8, Agilent column (5 μm, 300 Å, 4.6 × 250 mm).
Figure 5
Figure 5
Mass spectrum of J2-C3.
Figure 6
Figure 6
FT-IR spectrum of J2-C3.
Figure 7
Figure 7
Far-UV circular dichroism spectrum of J2-C3.
Figure 8
Figure 8
Inhibitory effect of J2-C3 on the proliferation of A549 cells.
See this image and copyright information in PMC

References

    1. Singh R., Sharma M., Joshi P., Rawat D.S. Clinical status of anti-cancer agents derived from marine sources. Anticancer Agents Med. Chem. 2008;8:603–617. - PubMed
    1. Schwartsmann G., Da Rocha A.B., Berlinck R., Jimeno J. Marine organisms as a source of new anticancer agents. Lancet Oncol. 2001;2:221–225. doi: 10.1016/S1470-2045(00)00292-8. - DOI - PubMed
    1. Suárez Y., González L., Cuadrado A., Berciano M., Lafarga M., Muñoz A. Kahalalide F, a new marine-derived compound, induces oncosis in human prostate and breast cancer cells. Mol. Cancer Ther. 2003;2:863–872. - PubMed
    1. Riely G.J., Gadgeel S., Rothman I., Saidman B., Sabbath K., Feit K., Kris M.G., Rizvi N.A. A phase 2 study of TZT-1027, administered weekly to patients with advanced non-small cell lung cancer following treatment with platinum-based chemotherapy. Lung Cancer. 2007;55:181–185. doi: 10.1016/j.lungcan.2006.10.002. - DOI - PubMed
    1. Kerbrat P., Dieras V., Pavlidis N., Ravaud A., Wanders J., Fumoleau P. Phase II study of LU 103793 (dolastatin analogue) in patients with metastatic breast cancer. Eur. J. Cancer. 2003;39:317–320. doi: 10.1016/S0959-8049(02)00531-2. - DOI - PubMed

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