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Cloning， Expression of Human CD147 and Its Effect on Matrix Metalloproteinases Expression of Fibroblast
Qing-Chuan Feng, Si-Jiao Chen1, Jin-Dan Song*
Department of Cell Biology, College of Basic Medical Sciences, China Medical University, Shenyang 110001, China;1Teaching and Research Office for Geriatric Disease of the First Affiliated Hospital of China Medical University, Shenyang 110001, C
Abstract: CD147， which stimulates nearby fibroblasts to synthesize matrix metalloproteinases (MMPs)， is associated with tumor cell invasion and metastasis. In order to investigate the role of CD147 on large colonic cancer， human cd147 gene was obtained from a normal individual by using RT-PCR， and was sequenced. One synonymic substitution and one nonsynonymic substitution were found in this gene. The nonsynonymic substitution C634T leads to a amino acid change (L212F)， which located in the transmembrane region of CD147. The bacterial and mammalian expression vectors of CD147 were constructed and cloned into E. coli BL-21 and CCL229 cells to induce expression of CD147， respectively. The results showed that the CD147 from BL-21/147 has a less molecular weight than from CCL229， suggesting the lack of glycosylation in CD147. The expression of CD147 was detected on the surface of CCL229/147 by using fluorescence microscope. The CD147 purified from CCL229/147 appeared to induce the production of MMPs in fibroblasts strongly， but that from BL-21/147 doesn't. Those results suggest that glycosylation may play a key role on the activity of CD147 to stimulate the expression of MMPs， and the mutation L212F does not influence the membrane location and activity of CD147 in CCL229.
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