ABSTRACT
AIM: To study the relationship between the CX3CL1 chemokine, its receptor CX3CR1, and gastric carcinoma/gastric carcinoma perineural invasion (PNI). METHODS: Thirty cases of gastric carcinoma were surgically resected (radical resection or palliative resection) between February 2012 and July 2012. Tumour and tumour-adjacent tissues were evaluated for the presence of CX3CL1 (ELISA) and CX3CR1 (immunohistochemistry and Western blotting) in an effort to analyse the relationship between CX3CL1/CX3CR1 and gastric carcinoma/gastric carcinoma PNI. RESULTS: Of these 30 cases, 14 were PNI-positive (46.7%). No significant differences in CX3CL and CX3CR1 expression in tumour-adjacent tissues were found between the PNI positive and negative groups. Expression levels of CX3CL and CX3CR1 in tumour tissues were significantly higher than those in adjacent tissues (P < 0.01), and were significantly higher in tumour tissues from the PNI-positive group compared to the PNI-negative group (P < 0.01). CONCLUSION: CX3CL1/CX3CR1 expression may be associated with the occurrence and development of gastric carcinoma as well as gastric carcinoma PNI.
Subject(s)
Carcinoma/metabolism , Chemokine CX3CL1/metabolism , Gene Expression Regulation, Neoplastic , Receptors, Chemokine/metabolism , Stomach Neoplasms/metabolism , Blotting, Western , CX3C Chemokine Receptor 1 , Carcinoma/pathology , Enzyme-Linked Immunosorbent Assay , Humans , Immunohistochemistry , Neoplasm Invasiveness , Neurons/pathology , Stomach Neoplasms/pathology , Treatment OutcomeABSTRACT
The full-length cDNA of Pinellia ternata agglutinin (PTA) was cloned from inflorescences using RACE-PCR. Through comparative analysis of PTA gene (pta) and its deduced amino acid sequence with those of other Araceae species, pta was found to encode a precursor lectin with signal peptide and to have extensive homology with those of other Araceae species. PTA was a heterotetrameric mannose-binding lectin with three mannose-binding boxes like lectins from other Araceae and Amaryllidaceae species. Southern blot analysis of the genomic DNA revealed that pta belonged to a low-copy gene family. Northern blot analysis demonstrated that pta constitutively expressed in various plant tissues including root, leaf, stem and inflorescence. The pta cDNA sequence encoding for mature PTA protein was cloned into pET-32a plasmid and the resulting plasmid, pET-32a-PTA containing Trx-PTA fusion protein, was investigated for the expression in E. coli BL21. SDS-PAGE gel analysis showed that the Trx-PTA fusion protein was successfully expressed in E. coli BL21 when induced by IPTG. Artificial diet assay revealed that PTA fusion protein had significant levels of resistance against peach potato aphids when incorporated into artificial diet at 0.1% (w/v). The cloning of the pta gene will enable us to further test its effect in depth on aphids by transferring the gene into crop plants.
Subject(s)
Plant Lectins/genetics , Amino Acid Sequence , Animals , Aphids/metabolism , Base Sequence , Cloning, Molecular , Models, Molecular , Molecular Sequence Data , Pinellia/genetics , Plant Lectins/chemistry , Plant Lectins/metabolism , Protein Structure, Secondary , Protein Structure, Tertiary , Recombinant Fusion Proteins/metabolism , Sequence AlignmentABSTRACT
A new lectin gene was cloned from Zephyranthes candida by using RACE-PCR. The full-length cDNA of Zephyranthes candida agglutinin (ZCA) was 647 bp and contained a 477 bp open reading frame encoding a 159 amino acid protein. Zephyranthes candida lectin gene was found to encode a precursor lectin with signal peptide and had extensive homology with those of other plant lectins. Molecular modeling of ZCA indicated that the three-dimensional structure of ZCA strongly resembles that of the snowdrop lectin, implying ZCA may have the similar insecticidal functions with GNA.