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Journal of Lipid Research, Vol. 42, 1897-1904, November 2001
Copyright © 2001 by Lipid Research, Inc.

Lipoprotein assembly capacity of the mammary tumor-derived cell line C127 is due to the expression of functional microsomal triglyceride transfer protein

Correspondence to: Gregory S. Shelness, To whom correspondence should be addressed., gshelnes{at}wfubmc.edu (E-mail)

C127, a murine mammary tumor-derived cell line, is capable of lipidating and secreting apolipoprotein B-41 (apoB-41) in the apparent absence of microsomal triglyceride transfer protein (MTP). Using a semiquantitative reverse transcriptase-coupled polymerase chain reaction, mouse MTP mRNA was detected in C127 cells at 10–20% of the relative abundance of human MTP in HepG2 cells. Radiolabeling of C127 cells with [³⁵S]methionine and [³⁵S]- cysteine followed by immunoprecipitation with anti-MTP antibodies identified a band with an electrophoretic mobility identical to that of authentic mouse MTP. Cotransfection of apoB-41 and the MTP 97-kDa subunit in C127 cells enhanced apoB secretion by 5-fold relative to apoB-41 transfection alone, suggesting that MTP is limiting in these cells. To establish that MTP expression is responsible for apoB-containing lipoprotein assembly in C127 cells, the effects of the MTP inhibitor BMS-200150 were examined. Secretion of apoB-41 by C127 cells was inhibited to the same extent observed in COS-1 cells cotransfected with apoB-41 and MTP.

These results suggest that low MTP expression, and not the expression or overexpression of another known or novel factor(s), is responsible for apoB assembly and secretion in C127 cells and further supports the essential nature of MTP in the biogenesis of apoB-containing lipoproteins. — Sellers, J. A., and G. S. Shelness. Lipoprotein assembly capacity of the mammary tumor-derived cell line C127 is due to the expression of functional microsomal triglyceride transfer protein. J. Lipid Res. 2001. 42: 1897–1904.

Supplementary key words: endoplasmic reticulum, MTP inhibitors, protein disulfide isomerase, protein secretion, very low density lipoprotein

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