Huh-7 or HepG2 Cells: Which is the Better Model for Studying Human Apolipoprotein-B100 Assembly and Secretion?

Overview

Journal J Lipid Res

Publisher Elsevier

Specialty Biochemistry

Date 2010 Oct 20

PMID 20956548

Citations 58

Authors

Steven J R Meex

Ursula Andreo

Janet D Sparks

Edward A Fisher

Affiliations

Soon will be listed here.

Abstract

Apolipoprotein-B100 (apoB100) is the essential protein for the assembly and secretion of very low density lipoproteins (VLDL) from liver. The hepatoma HepG2 cell line has been the cell line of choice for the study of synthesis and secretion of human apoB-100. Despite the general use of HepG2 cells to study apoB100 metabolism, they secrete relatively dense, lipid-poor particles compared with VLDL secreted in vivo. Recently, Huh-7 cells were adopted as an alternative model to HepG2 cells, with the implicit assumption that Huh-7 cells were superior in some respects of lipoprotein metabolism, including VLDL secretion. In this study we addressed the hypothesis that the spectrum of apoB100 lipoprotein particles secreted by Huh-7 cells more closely resembles the native state in human liver. We find that Huh-7 cells resemble HepG2 cells in the effects of exogenous lipids, microsomal triglyceride transfer protein (MTP)-inhibition, and proteasome inhibitors of apoB100 secretion, recovery, and degradation. In contrast to HepG2 cells, however, MEK-ERK inhibition does not correct the defect in VLDL secretion. Huh-7 cells do not appear to offer any advantages over HepG2 cells as a general model of human apoB100-lipoprotein metabolism.

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References

Ohsaki Y, Cheng J, Fujita A, Tokumoto T, Fujimoto T . Cytoplasmic lipid droplets are sites of convergence of proteasomal and autophagic degradation of apolipoprotein B. Mol Biol Cell. 2006; 17(6):2674-83. PMC: 1474802. DOI: 10.1091/mbc.e05-07-0659. View

Ginsberg H, Fisher E . The ever-expanding role of degradation in the regulation of apolipoprotein B metabolism. J Lipid Res. 2008; 50 Suppl:S162-6. PMC: 2674708. DOI: 10.1194/jlr.R800090-JLR200. View

Fisher E, Ginsberg H . Complexity in the secretory pathway: the assembly and secretion of apolipoprotein B-containing lipoproteins. J Biol Chem. 2002; 277(20):17377-80. DOI: 10.1074/jbc.R100068200. View

Pan M, Fisher E, Ginsberg H . The late addition of core lipids to nascent apolipoprotein B100, resulting in the assembly and secretion of triglyceride-rich lipoproteins, is independent of both microsomal triglyceride transfer protein activity and new triglyceride synthesis. J Biol Chem. 2001; 277(6):4413-21. DOI: 10.1074/jbc.M107460200. View

Kummrow E, Hussain M, Pan M, Marsh J, Fisher E . Myristic acid increases dense lipoprotein secretion by inhibiting apoB degradation and triglyceride recruitment. J Lipid Res. 2002; 43(12):2155-63. DOI: 10.1194/jlr.m200249-jlr200. View

Higashi Y, Itabe H, Fukase H, Mori M, Fujimoto Y, Takano T . Transmembrane lipid transfer is crucial for providing neutral lipids during very low density lipoprotein assembly in endoplasmic reticulum. J Biol Chem. 2003; 278(24):21450-8. DOI: 10.1074/jbc.M301376200. View

Lalanne F, Lambert G, Amar M, Chetiveaux M, Zair Y, Jarnoux A . Wild-type PCSK9 inhibits LDL clearance but does not affect apoB-containing lipoprotein production in mouse and cultured cells. J Lipid Res. 2005; 46(6):1312-9. DOI: 10.1194/jlr.M400396-JLR200. View

Fisher E, Zhou M, Mitchell D, Wu X, Omura S, Wang H . The degradation of apolipoprotein B100 is mediated by the ubiquitin-proteasome pathway and involves heat shock protein 70. J Biol Chem. 1997; 272(33):20427-34. DOI: 10.1074/jbc.272.33.20427. View

Wetterau J, GREGG R, Harrity T, Arbeeny C, Cap M, Connolly F . An MTP inhibitor that normalizes atherogenic lipoprotein levels in WHHL rabbits. Science. 1998; 282(5389):751-4. DOI: 10.1126/science.282.5389.751. View

10.

Pan M, Cederbaum A, Zhang Y, Ginsberg H, Williams K, Fisher E . Lipid peroxidation and oxidant stress regulate hepatic apolipoprotein B degradation and VLDL production. J Clin Invest. 2004; 113(9):1277-87. PMC: 398425. DOI: 10.1172/JCI19197. View

11.

Oyadomari S, Yun C, Fisher E, Kreglinger N, Kreibich G, Oyadomari M . Cotranslocational degradation protects the stressed endoplasmic reticulum from protein overload. Cell. 2006; 126(4):727-39. DOI: 10.1016/j.cell.2006.06.051. View

12.

McQueen M, Hawken S, Wang X, Ounpuu S, Sniderman A, Probstfield J . Lipids, lipoproteins, and apolipoproteins as risk markers of myocardial infarction in 52 countries (the INTERHEART study): a case-control study. Lancet. 2008; 372(9634):224-33. DOI: 10.1016/S0140-6736(08)61076-4. View

13.

Ohsaki Y, Cheng J, Suzuki M, Fujita A, Fujimoto T . Lipid droplets are arrested in the ER membrane by tight binding of lipidated apolipoprotein B-100. J Cell Sci. 2008; 121(Pt 14):2415-22. DOI: 10.1242/jcs.025452. View

14.

Brodsky J, Fisher E . The many intersecting pathways underlying apolipoprotein B secretion and degradation. Trends Endocrinol Metab. 2008; 19(7):254-9. PMC: 3216472. DOI: 10.1016/j.tem.2008.07.002. View

15.

Mitchell D, Zhou M, Pariyarath R, Wang H, AITCHISON J, Ginsberg H . Apoprotein B100 has a prolonged interaction with the translocon during which its lipidation and translocation change from dependence on the microsomal triglyceride transfer protein to independence. Proc Natl Acad Sci U S A. 1998; 95(25):14733-8. PMC: 24518. DOI: 10.1073/pnas.95.25.14733. View

16.

Yeung S, Chen S, Chan L . Ubiquitin-proteasome pathway mediates intracellular degradation of apolipoprotein B. Biochemistry. 1996; 35(43):13843-8. DOI: 10.1021/bi9618777. View

17.

Higashi Y, Itabe H, Fukase H, Mori M, Fujimoto Y, Sato R . Distribution of microsomal triglyceride transfer protein within sub-endoplasmic reticulum regions in human hepatoma cells. Biochim Biophys Acta. 2002; 1581(3):127-36. DOI: 10.1016/s1388-1981(02)00157-9. View

18.

Tsai J, Qiu W, Kohen-Avramoglu R, Adeli K . MEK-ERK inhibition corrects the defect in VLDL assembly in HepG2 cells: potential role of ERK in VLDL-ApoB100 particle assembly. Arterioscler Thromb Vasc Biol. 2006; 27(1):211-8. DOI: 10.1161/01.ATV.0000249861.80471.96. View

19.

Benoist F, Grand-Perret T . Co-translational degradation of apolipoprotein B100 by the proteasome is prevented by microsomal triglyceride transfer protein. Synchronized translation studies on HepG2 cells treated with an inhibitor of microsomal triglyceride transfer protein. J Biol Chem. 1997; 272(33):20435-42. DOI: 10.1074/jbc.272.33.20435. View

20.

Zhou M, Fisher E, Ginsberg H . Regulated Co-translational ubiquitination of apolipoprotein B100. A new paradigm for proteasomal degradation of a secretory protein. J Biol Chem. 1998; 273(38):24649-53. DOI: 10.1074/jbc.273.38.24649. View