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Amino acid sequence for inhibiting ptx3 to treat nasopharyngeal carcinoma

a nasopharyngeal carcinoma and ptx3 technology, applied in the direction of peptide/protein ingredients, peptide sources, drug compositions, etc., can solve the problems of poor prognosis and high levels of macrophage infiltration into tumors, and achieve the effects of inhibiting macrophage phagocytosis, promoting migration and invasion, and promoting angiogenesis

Inactive Publication Date: 2014-12-11
NAT CHENG KUNG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invention provides an amino acid sequence that can inhibit the migration, invasion, angiogenesis, and impair the phagocytosis of nasopharyngeal carcinoma cells. This amino acid sequence can be used to treat nasopharyngeal carcinoma.

Problems solved by technology

Clinical investigations have shown that high levels of macrophage infiltration into tumors are associated with a poor prognosis.

Method used

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  • Amino acid sequence for inhibiting ptx3 to treat nasopharyngeal carcinoma
  • Amino acid sequence for inhibiting ptx3 to treat nasopharyngeal carcinoma
  • Amino acid sequence for inhibiting ptx3 to treat nasopharyngeal carcinoma

Examples

Experimental program
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Effect test

example 1

Cell Culture and Treatment

[0033]Different cell lines including THP-1, NPC-TW01 [TW01] and HONE-1 were cultured in RPMI-1640 medium (Hyclone) containing 10% fetal bovine serum (FBS), 100 μg / ml streptomycin, and 100 units / ml penicillin. A mouse lung cancer cell line LLC1 was maintained in DMEM medium supplemented with 10% FBS, 100 μg / ml streptomycin and 100 units / ml penicillin. Mouse bone marrow mononuclear cells were obtained from the femur and tibia of mice and grown in RPMI-1640 medium containing 10% FBS and 25 ng / mL M-CSF (R&D systems Inc.). For mouse macrophage differentiation, mouse bone marrow mononuclear cells were grown in medium containing M-CSF and allowed to adhere. The adherent bone marrow-purified macrophages were then re-seeded in RPMI-1640 medium with 10% FBS. In this study, the dosage of PGE2 (Sigma) applied in every treatment is 15 ng / ml.

example 2

Phagocytosis Assay of euPTX3 Purified from Mammals

[0034]HONE-1 or TW01 were stained with a PKH-26 red fluorescent cell linker kit (Sigma) according to the manufacturer's instructions. 5×104 PKH-26-stained NPC cells were then treated with 300 and 600 ng / ml euPTX3 (purified from eukaryotic mouse myeloma cells, R&D Systems Inc.) or conditioned medium. After removing the PTX3 protein or conditioned medium, the experimental cells were reseeded and co-cultured with PMA-treated PKH-67-stained THP-1 cells for 3 h (the human monocyte cell line, THP-1 cells, can differentiate to macrophages by treating with PMA) and fluorescence signals were analyzed by flow cytometry. Phagocytosis activity was expressed as the percentage of green+ / red+ dual-fluorescent cells in the fluorescein isothiocyanate (FITC+) single-fluorescent cell population. The results as shown in FIG. 1, euPTX3 attenuates NPC cells (HONE-1 and TW01) phagocytosed by activated macrophages. Furthermore, the level of attenuation is d...

example 3

Analyzing the Effect of PTX3 / Ptx3 Purified from Mammals on Macrophage Phagocytosis Activity

[0035]At first, the conditional media were harvested from 1×106 stable THP-1 cells (with pCDNA3 / HA backbone vector) or 1×106 stable CEBPD-expressing THP-1 cells (with pCDNA3 / HA-CEBPD vector). The secreted PTX3 in conditioned media of above experimental cells were detected by human PTX3 ELISA Kit (R&D systems Inc.). As shown in FIG. 2-A, PTX3 is increased in the conditioned medium of CEBPD-expressing THP-1 cells.

[0036]After neutralizing PTX3 in conditioned medium with 1 ug / ml control antibody or PTX3 antibody (ab90807, Abcam) for 4 hours, 5×104 HONE-1 or 5×104 TW01 cells were co-cultured with PMA-treated PKH-67-stained THP-1 cells in above neutralized conditioned media for 3 hours. Phagocytosis activity was analyzed and expressed as the percentage of green+ / red+ dual-fluorescent cells in the FITC+ single-fluorescent cell population. As shown in FIG. 2-B, PTX3 antibody inhibits CEBPD-suppressed ...

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Abstract

The invention relates to an amino acid sequence for inhibiting PTX3 to treat nasopharyngeal carcinoma. It can be used to inhibit PTX3 from promoting the migration and invasion of nasopharyngeal carcinoma cells, promoting angiogenesis and inhibiting macrophage phagocytosis to further treat nasopharyngeal carcinoma.

Description

BACKGROUND OF THE INVENTION[0001]1. Field of the Invention[0002]The present invention relates to an amino acid sequence for inhibiting PTX3 to treat nasopharyngeal carcinoma (NPC), more particularly for inhibition of PTX3 from promoting the migration and invasion of nasopharyngeal carcinoma cells, promoting angiogenesis and inhibiting macrophage phagocytosis to further treat nasopharyngeal carcinoma.[0003]2. Description of Related Art[0004]Many researches reported that acute inflammation increases the probabilities of normal cells to become tumorigenic and enhances the occurring rate of cancer cell migration and invasion. Tumor-associated macrophages (TAM) are the most abundant immune cells within the tumor stroma and are required for a number of functions important for tumor progression, such as promoting tumor cell proliferation, angiogenesis, incessant matrix turnover and repressing the adaptive immune responses. Clinical investigations have shown that high levels of macrophage i...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C07K14/435
CPCC07K14/435A61K38/00C07K14/47A61K38/16A61K38/1709A61P35/00C07K14/71Y02A50/30
Inventor WANG, JU-MINGHSIAO, YU-WEI
Owner NAT CHENG KUNG UNIV
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