The Transmembrane Conductance Mechanism Of A Peptide With High Affinity To Human Breast Cancer And Analysis Membrane Protein Structure Differences

Objective:Detection cells affinity of PI and MDA-MB-231 breast cancer cells; To investigate transmembrane transduction mechanism of a peptide (PI) with high affinity to human breast cancer cell line MDA-MB-231; two-dimensional electrophoresis analysis of MDA-MB-231 breast cancer cells and Calu-1 lung cancer cell membrane structure differences; For the future Continue to study the specificity of PI transduction mechanism of transmembrane experimental basis.Methods:PI were syntheticed and labeled FITC green fluorescence in its N-terminus. Setting human lung cancer cells Calu-1 and human breast cancer cells MDA-MB-435 as a positive control and negative control cells, Observing FITC-PI distribution in the cell in Vitro; Adding FITC-PI In the culture system that human breast cancer cells MDA-MB-231 were incubated with MHC-I antibody, Observing the affinity of FITC-PI and MDA-MB-231 Cell which MHC-I molecule of cell surface were closed by MHC-I antibody under fluorescence microscopy. Selectted Calul cells which the also have high affinity with the FITC-PI as a positive control, HLA-allele analysis was performed by polymerase chain reaction-sequence specific primers., To investigate whether the transmembrane transduction mechanism of PI was associated with the MHC-I antigen; the distribution of FITC-PI in MDA-MB-231 cells were detected using inverted fluorescence microscope after pre-incubation with an inhibitor of endocytosis,nystatin-To investigate whether the Transmembrane transduction mechanism of PI isassociated with the caveolin-mediated endocytosis. To investigate whether the transmembrane transduction mechanism of PI is associated with the same membrane protein of the two cell lines, the membrane proteins were detected by two-dimensional electrophoresis.Results:FITC-PI was observed in the cytoplasm and nuclei of MDA-MB-231 breast cancer cells and calu-1 cells, MDA-MB-435 breast cancer cells have nothing, FITC-PI was observed in the two cells with MHC-I antigen being blocked.Gene typing-analysis revealed that the alleles of the HLA-A and B locis were different. The inhibitor nystatin suppressed FITC-PI distribution. Two-dimensional electrophoresis analysis showed that membrane protein profiles of the two cell lines had 11 same protein spots.Conclusion:The transduction of PI is mediated by the caveolin-mediated endocytosis.The same proteins of the two cell lines may be also involved in this process.This study provides experimental evidence for further investigation of the transmembrane transduction mechanism.