ABSTRACT
Objective To observe the expression of Neuropilin (NRP)-1, 2, to calculate micro vessel density (MVD) in colorectal cancer, paraneoplastic tissues, normal tissues, and to investigate the correlation between NRP-1, 2 and MVD to understand the role of NRP-1, 2 in the process of tumor angiogenesis. Methods Expression of NRP-1 and NRP-2 were studied by immunohistochemistry in 66 specimens from colorectal cancer, paraneoplastic tissues and normal tissues. MVD was assessed based on CD105 immunohistochical staining. Results (1) The positive expression of NRP-1, 2 in colorectal cancer, paraneoplastic tissues, normal tissues were 71.2%, 25.8%, 0;80.3%, 15.2%, 0 respectively. There was a statistical difference between them (P0.05). (4) The MVD value had no correlation with the gender or age of patients, tumor size, position, histological types (P>0.05), but had a good correlation with infiltrating depth, lymphatic metastasis and Dukes staging (P<0.05). (5) There was a positive correlation between NRP-1, 2 and MVD value (P<0.01). Conclusion NRP-1, 2 may play an important role in the angiogenesis of the colorectal cancer, and it correlates with the invasion and metastasis of colorectal cancer closely.
ABSTRACT
Objective To construct Neuropilins-2 eukaryotic expression vector for RNA interference.Methods Recombinant targeting on gene NRP2 was designed and established with plasmid pGenSil-1 based on NRP2 cDNA equences of Genomes.Two pairs of oligonucleotides were synthesized according to the Tuschl and inserted into plasmid pGenSil-l to generate siRNA eukaryotic expression vector,DH5? strains were transformed,plasmid were extracted,and recombinant vectors were identified by the restriction map and the sequence analysis.The recombinant plasmid(pGenSil-NRP2) was transfected into the cultured LOVO cells.At 48 h after transfection,the whole cell protein was extracted,and the protein level was detected by Western blotting with mouse-anti-human NRP2 monoclonal antibody.Results Recombinant plasmids were completely coincided with the designs by the restriction map and the sequence analysis.pGenSil-NRP2 expression vector into LOVO cells down-regulated the protein level of NRP2 at 48 h after transfection.The recombinant eukaryotic expression vector were constructed successfully.Conclusion siRNA recombinant can be constructed successfully by RNAi technique for inhibiting NRP2 expression.