Abstract
This paper describes a method, designated "receptor-dependent photosensitization," by which the receptor-mediated endocytosis of low density lipoprotein (LDL) can be used to deliver a photosensitizing agent, pyrene, to cultured human and animal cells. The hydrophobic core of LDL is extracted and replaced with pyrene covalently coupled to cholesteryl oleate. This reconstituted LDL enters cells in significant amounts only when the cells express LDL receptors, resulting in the accumulation of pyrene cholesteryl oleate within lysosomes. Subsequent exposure of the cells to ultraviolet light leads to cell death. Cells killed by this technique include normal and simian virus 40-transformed human fibroblasts, human A-431 epidermal carcinoma cells, Chinese hamster ovary cells, and mouse L cells, all of which express LDL receptors. Mutant fibroblasts from a patient with homozygous familial hypercholesterolemia, which lack LDL receptors, do not take up significant amounts of the pyrene-containing LDL and are not killed by subsequent exposure to light. The current experiments establish the feasibility of receptor-dependent photosensitization as an efficient and selective method for killing cultured human and animal cells.
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- Allison A. C., Magnus I. A., Young M. R. Role of lysosomes and of cell membranes in photosensitization. Nature. 1966 Feb 26;209(5026):874–878. doi: 10.1038/209874a0. [DOI] [PubMed] [Google Scholar]
- Anderson R. G., Brown M. S., Goldstein J. L. Inefficient internalization of receptor-bound low density lipoprotein in human carcinoma A-431 cells. J Cell Biol. 1981 Feb;88(2):441–452. doi: 10.1083/jcb.88.2.441. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Basu S. K., Goldstein J. L., Anderson R. G., Brown M. S. Monensin interrupts the recycling of low density lipoprotein receptors in human fibroblasts. Cell. 1981 May;24(2):493–502. doi: 10.1016/0092-8674(81)90340-8. [DOI] [PubMed] [Google Scholar]
- Brown M. S., Dana S. E., Goldstein J. L. Regulation of 3-hydroxy-3-methylglutaryl coenzyme A reductase activity in cultured human fibroblasts. Comparison of cells from a normal subject and from a patient with homozygous familial hypercholesterolemia. J Biol Chem. 1974 Feb 10;249(3):789–796. [PubMed] [Google Scholar]
- Cherington P. V., Smith B. L., Pardee A. B. Loss of epidermal growth factor requirement and malignant transformation. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3937–3941. doi: 10.1073/pnas.76.8.3937. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Diamond I., Granelli S. G., McDonagh A. F., Nielsen S., Wilson C. B., Jaenicke R. Photodynamic therapy of malignant tumours. Lancet. 1972 Dec 2;2(7788):1175–1177. doi: 10.1016/s0140-6736(72)92596-2. [DOI] [PubMed] [Google Scholar]
- Goldstein J. L., Anderson R. G., Brown M. S. Coated pits, coated vesicles, and receptor-mediated endocytosis. Nature. 1979 Jun 21;279(5715):679–685. doi: 10.1038/279679a0. [DOI] [PubMed] [Google Scholar]
- Goldstein J. L., Brown M. S., Stone N. J. Genetics of the LDL receptor: evidence that the mutations affecting binding and internalization are allelic. Cell. 1977 Nov;12(3):629–641. doi: 10.1016/0092-8674(77)90263-x. [DOI] [PubMed] [Google Scholar]
- Goldstein J. L., Brown M. S. The low-density lipoprotein pathway and its relation to atherosclerosis. Annu Rev Biochem. 1977;46:897–930. doi: 10.1146/annurev.bi.46.070177.004341. [DOI] [PubMed] [Google Scholar]
- Goldstein J. L., Helgeson J. A., Brown M. S. Inhibition of cholesterol synthesis with compactin renders growth of cultured cells dependent on the low density lipoprotein receptor. J Biol Chem. 1979 Jun 25;254(12):5403–5409. [PubMed] [Google Scholar]
- Ho Y. K., Smith R. G., Brown M. S., Goldstein J. L. Low-density lipoprotein (LDL) receptor activity in human acute myelogenous leukemia cells. Blood. 1978 Dec;52(6):1099–1114. [PubMed] [Google Scholar]
- Krieger M., Brown M. S., Faust J. R., Goldstein J. L. Replacement of endogenous cholesteryl esters of low density lipoprotein with exogenous cholesteryl linoleate. Reconstitution of a biologically active lipoprotein particle. J Biol Chem. 1978 Jun 25;253(12):4093–4101. [PubMed] [Google Scholar]
- Krieger M., Goldstein J. L., Brown M. S. Receptor-mediated uptake of low density lipoprotein reconstituted with 25-hydroxycholesteryl oleate suppresses 3-hydroxy-3-methylglutaryl-coenzyme A reductase and inhibits growth of human fibroblasts. Proc Natl Acad Sci U S A. 1978 Oct;75(10):5052–5056. doi: 10.1073/pnas.75.10.5052. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Krieger M., Smith L. C., Anderson R. G., Goldstein J. L., Kao Y. J., Pownall H. J., Gotto A. M., Jr, Brown M. S. Reconstituted low density lipoprotein: a vehicle for the delivery of hydrophobic fluorescent probes to cells. J Supramol Struct. 1979;10(4):467–478. doi: 10.1002/jss.400100409. [DOI] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Moan J., Pettersen E. O., Christensen T. The mechanism of photodynamic inactivation of human cells in vitro in the presence of haematoporphyrin. Br J Cancer. 1979 Apr;39(4):398–407. doi: 10.1038/bjc.1979.72. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schothorst A. A., Suurmond D., Ploem J. S. In vitro studies on the protoporphyrin uptake and photosensitivity of normal skin fibroblasts and fibroblasts from patients with erythropoietic protoporphyria. J Invest Dermatol. 1977 Dec;69(6):551–557. doi: 10.1111/1523-1747.ep12688381. [DOI] [PubMed] [Google Scholar]
- Slater T. F., Riley P. A. Photosensitization and lysosomal damage. Nature. 1966 Jan 8;209(5019):151–154. doi: 10.1038/209151a0. [DOI] [PubMed] [Google Scholar]
- Tomson S. H., Emmett E. A., Fox S. H. Photodestruction of mouse epithelial tumors after oral acridine orange and argon laser. Cancer Res. 1974 Nov;34(11):3124–3127. [PubMed] [Google Scholar]
- Weisgraber K. H., Innerarity T. L., Mahley R. W. Role of lysine residues of plasma lipoproteins in high affinity binding to cell surface receptors on human fibroblasts. J Biol Chem. 1978 Dec 25;253(24):9053–9062. [PubMed] [Google Scholar]