Human Large Lung Research Articles

EXPRESSION AND PURIFICATION OF HUMAN ARP1 PROTEIN AND RAPID PREPARATION OF POLYCLONAL ANTIBODY

Angiopoietin-related protein 1 (ARP1) is one of the antiangiogenic factors and plays an important role in endothelial cell proliferation, migration, and blood vessel network formation. Here a rapid method to prepare ARP1 polyclonal antibody in 1 month was developed. The gene of fibrinogen homology domain (FD) for ARP1 was cloned and the protein was expressed in a soluble form of MBP-FD fused protein. The MBP-FD protein was purified using amylose affinity chromatography of maltose-binding protein. Polyclonal antibodies against MBP-FD were obtained through immunization in BALB/c mice. The titer was determined by indirect enzyme-linked immunosorbent assay (ELISA), and the antibody specificity was assessed by Western blot. The full-length ARP1 protein in stable form expressed in transfected human large lung cancer cell lines NCI-H460 was detected by immunocytochemistry (ICC) analysis using ARP1 polyclonal antibodies. The result shows that the antibody possesses good specificity and sensitivity. This work provides a substantial base for the further studies of ARP1 function and associated mechanisms. Supplemental materials are available for this article. Go to the publisher's online edition of Preparative Biochemistry and Biotechnology to view the supplemental file.

Research on the anti-tumor effect of Plumbagin against human large cell lung cancer cell line NL9980

Objective To investigate the anti-tumor effect and mechanism of Plumbagin against human large cell lung cancer cell line NL9980.Methods Plumbagin in different concentrations were set up to administrate NL9980 cell line.MTT test was performed to detect the inhibiting effects against tumor proliferation and IC50 concentration was identified.The influence of tumor apoptosis was observed by flow cytometry and the inhibiting effect against invasive ability was detected by Boyden chamber test.IC50 group and control group were set up and tumor cells were harvested 6 h,24 h and 48 h after administration.Realtime PCR was used to detect mRNA expression changes of bcl-2,bax,VEGF and CYCD1.Results MTT test showed obvious inhibiting effects of proliferation in NL9980 cell line and IC50 was 7.5 μmol/L.Plumbagin induced the apoptosis of tumor cells and showed a significant anti-invasive effect.Mean membrane-spanning cells were 161.59±47.32 and 26.58±9.07 in control group and IC50 group correspondingly.Down-regulations of bcl-2,VEGF and CYCD1 were detected and expression of bax gene increased.The effect of Plumbagin showed obvious time dependance (P ＜0.05).Conclusion Plumbagin has displayed the powerful antitumor effects against NL9980 cell line and it might work via multiple pathways.Plumbagin exhibits the prospect of being an effective drug against large cell lung cancer. Key words: Lung neoplasms; Carcinoma, large cell; Plumbagin; Proliferation; Invasion; Apoptosis