Cloning,transfer and anti-human breast cancer cell growth of the extracellular domain of cadherin 5 / 中国病理生理杂志

ABSTRACT

AIM:In order to study the effect of the extracellular domain of cadherin 5 on the growth of a human breast cancer cell line MDA-MB435.METHODS:Cadherin extracellular domain repeats 1 to 4(CED1-4)was cloned by using RT-PCR technique,and inserted into the plasmid vector pMSCV.pMSCV-CED1-4 was propagated in XL-blue strain of Escherichia coli,extracted and purified.CED1-4 was cut by restriction endonuclease,examined by using agar gel electrophoresis,and finally sequenced.CED1-4 gene was transferred into MDA-MB435 cell line.The expression of CED1-4 gene in MDA-MB435 cell was analyzed by methods of RT-PCR and Western blotting.The effect of CED1-4 on the growth of MDA-MB435 cell was observed by the methods of proliferation experiments in vitro and the experiments in nude mice in vivo.RESULTS:The recombinant vector pMSCV-CED1-4 was successfully constructed.CED1-4 band appeared between the 1 636 bp and 1 018 bp in agar gel electrophoresis.The sequence result showed that CED1-4 had 1 452 bp and codes 484 amino acids.PCR and Western blotting identified that CED1-4 mRNA and protein were expressed in the transfected MDA-MB435 cells.Cell proliferation experiments showed that the proliferation rate of MDA-MB435 cells was lower in the experimental group than that in the experimental control group and the blank control group.The mean volume and weight of tumors in nude mice were lower in the experimental group than those in the experimental control group and the blank control group.CONCLUSION:The growth of a human breast cancer cell line MDA-MB435 is inhibited in vitro and in vivo by cadherin 5 extracellular domain CED1-4.