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. 1985 Dec;5(12):3552–3559. doi: 10.1128/mcb.5.12.3552

Effect of dimethyl sulfoxide on human carcinoma cells, inhibition of plasminogen activator synthesis, change in cell morphology, and alteration of response to cholera toxin.

L Ossowski, D Belin
PMCID: PMC369186  PMID: 3837848

Abstract

Human carcinoma HEp-3 lost its tumorigenic and metastatic potential upon prolonged culture in vitro. This change was accompanied by a reduced production of plasminogen activator (PA) of the urokinase type (uPA), which is secreted by HEp-3 cells, a change in response to effectors that modulate uPA production, and an alteration of cell morphology. Similar but more rapid changes occurred when malignant HEp-3 cells were exposed to dimethyl sulfoxide (DMSO). uPA activity in the culture medium dropped below 50% of the control level within 6 h after the addition of DMSO and became undetectable after 24 h of treatment. This drop in uPA activity was not caused by an increased production of PA inhibitors. The cell-associated uPA decreased to 25 to 30% of the control level within 6 h of DMSO treatment and remained at this level for at least 96 h; the reduced uPA production was partially accounted for by a rapid decrease in the functional and chemical concentration of uPA mRNA. In contrast, the concentrations of most of the abundant mRNA species did not appear to be significantly affected, and cell growth was only slightly inhibited in the presence of DMSO. Malignant HEp-3 cells treated with DMSO responded to cholera toxin with an enhanced production of uPA, and their morphology became indistinguishable from that of nonmalignant HEp-3 cells grown in vitro for prolonged periods of time. All of the above changes were fully and rapidly reversible. The inhibitory effect of DMSO on PA production appears to be specific for uPA of human cell lines.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Belin D., Godeau F., Vassalli J. D. Tumor promoter PMA stimulates the synthesis and secretion of mouse pro-urokinase in MSV-transformed 3T3 cells: this is mediated by an increase in urokinase mRNA content. EMBO J. 1984 Aug;3(8):1901–1906. doi: 10.1002/j.1460-2075.1984.tb02065.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Belin D., Vassalli J. D., Combépine C., Godeau F., Nagamine Y., Reich E., Kocher H. P., Duvoisin R. M. Cloning, nucleotide sequencing and expression of cDNAs encoding mouse urokinase-type plasminogen activator. Eur J Biochem. 1985 Apr 15;148(2):225–232. doi: 10.1111/j.1432-1033.1985.tb08829.x. [DOI] [PubMed] [Google Scholar]
  3. Brock M. L., Shapiro D. J. Estrogen stabilizes vitellogenin mRNA against cytoplasmic degradation. Cell. 1983 Aug;34(1):207–214. doi: 10.1016/0092-8674(83)90151-4. [DOI] [PubMed] [Google Scholar]
  4. Butler W. B., Kirkland W. L., Jorgensen T. L. Induction of plasminogen activator by estrogen in a human breast cancer cell line (MCF-7). Biochem Biophys Res Commun. 1979 Oct 29;90(4):1328–1334. doi: 10.1016/0006-291x(79)91181-1. [DOI] [PubMed] [Google Scholar]
  5. Camiolo S. M., Markus G., Englander L. S., Siuta M. R., Hobika G. H., Kohga S. Plasminogen activator content and secretion in explants of neoplastic and benign human prostate tissues. Cancer Res. 1984 Jan;44(1):311–318. [PubMed] [Google Scholar]
  6. Camiolo S. M., Markus G., Evers J. L., Hobika G. H., DePasquale J. L., Beckley S., Grimaldi J. P. Plasminogen activator content of neoplastic and benign human prostate tissues; fibrin augmentation of an activator activity. Int J Cancer. 1981 Feb 15;27(2):191–198. doi: 10.1002/ijc.2910270211. [DOI] [PubMed] [Google Scholar]
  7. Christman J. K., Weich N., Schoenbrun B., Schneiderman N., Acs G. Hypomethylation of DNA during differentiation of Friend erythroleukemia cells. J Cell Biol. 1980 Aug;86(2):366–370. doi: 10.1083/jcb.86.2.366. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Churchward G., Belin D., Nagamine Y. A pSC101-derived plasmid which shows no sequence homology to other commonly used cloning vectors. Gene. 1984 Nov;31(1-3):165–171. doi: 10.1016/0378-1119(84)90207-5. [DOI] [PubMed] [Google Scholar]
  9. Corasanti J. G., Celik C., Camiolo S. M., Mittelman A., Evers J. L., Barbasch A., Hobika G. H., Markus G. Plasminogen activator content of human colon tumors and normal mucosae: separation of enzymes and partial purification. J Natl Cancer Inst. 1980 Aug;65(2):345–351. [PubMed] [Google Scholar]
  10. DAGG C. P., KARNOFSKY D. A., TOOLAN H. W., RODDY J. Serial passage of human tumors in chick embryo: growth inhibition by nitrogen mustard. Proc Soc Exp Biol Med. 1954 Oct;87(1):223–227. doi: 10.3181/00379727-87-21341. [DOI] [PubMed] [Google Scholar]
  11. Dairkee S. H., Glaser D. A. Dimethyl sulfoxide affects colony morphology on agar and alters distribution of glycosaminoglycans and fibronectin. Proc Natl Acad Sci U S A. 1982 Nov;79(22):6927–6931. doi: 10.1073/pnas.79.22.6927. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Deutsch D. G., Mertz E. T. Plasminogen: purification from human plasma by affinity chromatography. Science. 1970 Dec 4;170(3962):1095–1096. doi: 10.1126/science.170.3962.1095. [DOI] [PubMed] [Google Scholar]
  13. Dobberstein B., Blobel G. Functional interaction of plant ribosomes with animal microsomal membranes. Biochem Biophys Res Commun. 1977 Feb 21;74(4):1675–1682. doi: 10.1016/0006-291x(77)90637-4. [DOI] [PubMed] [Google Scholar]
  14. Friend C., Scher W., Holland J. G., Sato T. Hemoglobin synthesis in murine virus-induced leukemic cells in vitro: stimulation of erythroid differentiation by dimethyl sulfoxide. Proc Natl Acad Sci U S A. 1971 Feb;68(2):378–382. doi: 10.1073/pnas.68.2.378. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Granelli-Piperno A., Reich E. A study of proteases and protease-inhibitor complexes in biological fluids. J Exp Med. 1978 Jul 1;148(1):223–234. doi: 10.1084/jem.148.1.223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Granelli-Piperno A., Reich E. Plasminogen activators of the pituitary gland: enzyme characterization and hormonal modulation. J Cell Biol. 1983 Oct;97(4):1029–1037. doi: 10.1083/jcb.97.4.1029. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hoal E. G., Wilson E. L., Dowdle E. B. The regulation of tissue plasminogen activator activity by human fibroblasts. Cell. 1983 Aug;34(1):273–279. doi: 10.1016/0092-8674(83)90158-7. [DOI] [PubMed] [Google Scholar]
  18. Huberman E., Heckman C., Langenbach R. Stimulation of differentiated functions in human melanoma cells by tumor-promoting agents and dimethyl sulfoxide. Cancer Res. 1979 Jul;39(7 Pt 1):2618–2624. [PubMed] [Google Scholar]
  19. Jefferson D. M., Clayton D. F., Darnell J. E., Jr, Reid L. M. Posttranscriptional modulation of gene expression in cultured rat hepatocytes. Mol Cell Biol. 1984 Sep;4(9):1929–1934. doi: 10.1128/mcb.4.9.1929. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kimhi Y., Palfrey C., Spector I., Barak Y., Littauer U. Z. Maturation of neuroblastoma cells in the presence of dimethylsulfoxide. Proc Natl Acad Sci U S A. 1976 Feb;73(2):462–466. doi: 10.1073/pnas.73.2.462. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Laverriere J. N., Morin A., Tixier-Vidal A., Truong A. T., Gourdji D., Martial J. A. Inverse control of prolactin and growth hormone gene expression: effect of thyroliberin on transcription and RNA stabilization. EMBO J. 1983;2(9):1493–1499. doi: 10.1002/j.1460-2075.1983.tb01613.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Leavitt J., Barrett J. C., Crawford B. D., Ts'o P. O. Butyric acid suppression of the in vitro neoplastic state of Syrian hamster cells. Nature. 1978 Jan 19;271(5642):262–265. doi: 10.1038/271262a0. [DOI] [PubMed] [Google Scholar]
  23. Loskutoff D. J., van Mourik J. A., Erickson L. A., Lawrence D. Detection of an unusually stable fibrinolytic inhibitor produced by bovine endothelial cells. Proc Natl Acad Sci U S A. 1983 May;80(10):2956–2960. doi: 10.1073/pnas.80.10.2956. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Mak T. W., Rutledge G., Sutherland D. J. Androgen-dependent fibrinolytic activity in a murine mammary carcinoma (Shionogi SC-115 cells) in vitro. Cell. 1976 Feb;7(2):223–226. doi: 10.1016/0092-8674(76)90021-0. [DOI] [PubMed] [Google Scholar]
  25. Mangiarotti G., Ceccarelli A., Lodish H. F. Cyclic AMP stabilizes a class of developmentally regulated Dictyostelium discoideum mRNAs. Nature. 1983 Feb 17;301(5901):616–618. doi: 10.1038/301616a0. [DOI] [PubMed] [Google Scholar]
  26. Minty A. J., Birnie G. D., Paul J. Gene expression in Friend erythroleukemia cells following the induction of hemoglobin synthesis. Exp Cell Res. 1978 Aug;115(1):1–14. doi: 10.1016/0014-4827(78)90395-6. [DOI] [PubMed] [Google Scholar]
  27. Mira-y-Lopez R., Reich E., Ossowski L. Modulation of plasminogen activator in rodent mammary tumors by hormones and other effectors. Cancer Res. 1983 Nov;43(11):5467–5477. [PubMed] [Google Scholar]
  28. Miskin R., Soreq H. Microinjected Xenopus oocytes synthesize active human plasminogen activator. Nucleic Acids Res. 1981 Jul 24;9(14):3355–3363. doi: 10.1093/nar/9.14.3355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Moore G. E., Merrick S. B., Woods L. K., Arabasz N. M. A human squamous cell carcinoma cell line. Cancer Res. 1975 Oct;35(10):2684–2688. [PubMed] [Google Scholar]
  30. Ossowski L., Biegel D., Reich E. Mammary plasminogen activator: correlation with involution, hormonal modulation and comparison between normal and neoplastic tissue. Cell. 1979 Apr;16(4):929–940. doi: 10.1016/0092-8674(79)90108-9. [DOI] [PubMed] [Google Scholar]
  31. Ossowski L., Reich E. Antibodies to plasminogen activator inhibit human tumor metastasis. Cell. 1983 Dec;35(3 Pt 2):611–619. doi: 10.1016/0092-8674(83)90093-4. [DOI] [PubMed] [Google Scholar]
  32. Ossowski L., Reich E. Changes in malignant phenotype of a human carcinoma conditioned by growth environment. Cell. 1983 Jun;33(2):323–333. doi: 10.1016/0092-8674(83)90414-2. [DOI] [PubMed] [Google Scholar]
  33. Ossowski L., Reich E. Experimental model for quantitative study of metastasis. Cancer Res. 1980 Jul;40(7):2300–2309. [PubMed] [Google Scholar]
  34. Ossowski L., Reich E. Loss of malignancy during serial passage of human carcinoma in culture and discordance between malignancy and transformation parameters. Cancer Res. 1980 Jul;40(7):2310–2315. [PubMed] [Google Scholar]
  35. Ossowski L., Unkeless J. C., Tobia A., Quigley J. P., Rifkin D. B., Reich E. An enzymatic function associated with transformation of fibroblasts by oncogenic viruses. II. Mammalian fibroblast cultures transformed by DNA and RNA tumor viruses. J Exp Med. 1973 Jan 1;137(1):112–126. doi: 10.1084/jem.137.1.112. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Pennica D., Holmes W. E., Kohr W. J., Harkins R. N., Vehar G. A., Ward C. A., Bennett W. F., Yelverton E., Seeburg P. H., Heyneker H. L. Cloning and expression of human tissue-type plasminogen activator cDNA in E. coli. Nature. 1983 Jan 20;301(5897):214–221. doi: 10.1038/301214a0. [DOI] [PubMed] [Google Scholar]
  37. Rudland P. S., Davies A. T., Warburton M. J. Prostaglandin-induced differentiation or dimethyl sulfoxide-induced differentiation: reduction of the neoplastic potential of a rat mammary tumor stem-cell line. J Natl Cancer Inst. 1982 Nov;69(5):1083–1093. [PubMed] [Google Scholar]
  38. Scher W., Friend C. Breakage of DNA and alterations in folded genomes by inducers of differentiation in Friend erythroleukemic cells. Cancer Res. 1978 Mar;38(3):841–849. [PubMed] [Google Scholar]
  39. Shin S. I., Freedman V. H., Risser R., Pollack R. Tumorigenicity of virus-transformed cells in nude mice is correlated specifically with anchorage independent growth in vitro. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4435–4439. doi: 10.1073/pnas.72.11.4435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Sisskin E. E., Weinstein I. B., Evans C. H., Dipaolo J. A. Plasminogen activator synthesis accompanying chemical carcinogen-induced in vitro transformation of Syrian hamster and guinea-pig fetal cells. Int J Cancer. 1980 Sep 15;26(3):331–335. doi: 10.1002/ijc.2910260312. [DOI] [PubMed] [Google Scholar]
  41. Stoker M. G., Rubin H. Density dependent inhibition of cell growth in culture. Nature. 1967 Jul 8;215(5097):171–172. doi: 10.1038/215171a0. [DOI] [PubMed] [Google Scholar]
  42. Strickland S., Beers W. H. Studies on the role of plasminogen activator in ovulation. In vitro response of granulosa cells to gonadotropins, cyclic nucleotides, and prostaglandins. J Biol Chem. 1976 Sep 25;251(18):5694–5702. [PubMed] [Google Scholar]
  43. Unkeless J. C., Tobia A., Ossowski L., Quigley J. P., Rifkin D. B., Reich E. An enzymatic function associated with transformation of fibroblasts by oncogenic viruses. I. Chick embryo fibroblast cultures transformed by avian RNA tumor viruses. J Exp Med. 1973 Jan 1;137(1):85–111. doi: 10.1084/jem.137.1.85. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Vassalli J. D., Dayer J. M., Wohlwend A., Belin D. Concomitant secretion of prourokinase and of a plasminogen activator-specific inhibitor by cultured human monocytes-macrophages. J Exp Med. 1984 Jun 1;159(6):1653–1668. doi: 10.1084/jem.159.6.1653. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Verde P., Stoppelli M. P., Galeffi P., Di Nocera P., Blasi F. Identification and primary sequence of an unspliced human urokinase poly(A)+ RNA. Proc Natl Acad Sci U S A. 1984 Aug;81(15):4727–4731. doi: 10.1073/pnas.81.15.4727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Wilson E. L., Dowdle E. Secretion of plasminogen activator by normal, reactive and neoplastic human tissues cultured in vitro. Int J Cancer. 1978 Oct 15;22(4):390–399. doi: 10.1002/ijc.2910220405. [DOI] [PubMed] [Google Scholar]
  47. Wun T. C., Ossowski L., Reich E. A proenzyme form of human urokinase. J Biol Chem. 1982 Jun 25;257(12):7262–7268. [PubMed] [Google Scholar]

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