Skip to main content
Japanese Journal of Cancer Research : Gann logoLink to Japanese Journal of Cancer Research : Gann
. 1991 Jun;82(6):693–700. doi: 10.1111/j.1349-7006.1991.tb01905.x

Characterization of a 54 kDa, α1Antitrypsin‐like Protein Isolated from Ascitic Fluid of an Endometrial Cancer Patient

Naotake Tanaka 1, Souei Sekiya 1, Hiroyoshi Takamizawa 1, Noriko Kato 2, Yoichi Moriyama 2, Shinji Fujimura 2,
PMCID: PMC5918499  PMID: 1906855

Abstract

A protein factor which stimulated [3H]thymidine uptake into free hepatocytes prepared from normal mouse liver was detected in the ascitic fluid of gynecological cancer patients. The factor was subsequently further purified from the ascitic fluid of an endometrial cancer patient by DEAE‐Sephacel, Sephadex G‐150 and Phenyl‐Sepharose CL‐4B column chromatographies, and sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) showed a single protein band of 54,000 Da, designated tentatively as 54K ascitic protein (54K‐AP). 54K‐AP was similar to human Qt‐antitrypsin (α1‐AT) in terms of SDS‐PAGE and immunological behavior, but was slightly different in terms of amino acid sequence and isoelectric point. Although 54K‐AP inhibited the activities of bovine trypsin and a‐chymotrypsin as did human α1‐AT, 54K‐AP inhibited the plasminogen activator released from human endometrial cancer Ishikawa cells more efficiently than α1‐AT. Because, in contrast to normal serum, the serum from the endometrial cancer patients stimulated [3H]thymidine uptake into hepatocytes, the possibility arises that 54K‐AP could he produced hy the cancer host as a defence mechanism against the cancer.

Keywords: a1‐Antitrypsin‐like protein, Thymidine uptake, Endometrial cancer, Tumor host ascites

Full Text

The Full Text of this article is available as a PDF (534.1 KB).

REFERENCES

  • 1. ) Fujimura , S. and Shimizu , M.Enhanced activity of tRNA‐pseudouridine synthetase in Yoshida ascites sarcoma . Biochem. Biophys. Res. Commun. , 79 , 763 – 768 ( 1977. ). [DOI] [PubMed] [Google Scholar]
  • 2. ) Shimizu , M. and Fujimura , S.Studies on the abnormal excretion of pyrimidine nucleosides in the urine of Yoshida ascites sarcoma‐bearing rats. Increased excretion of deoxycytidine, pseudouridine and cytidine . Biochim. Biophys. Acta , 517 , 277 – 286 ( 1978. ). [DOI] [PubMed] [Google Scholar]
  • 3. ) Shimizu , M. , and Fujimura , S.Origin of increased deoxycytidine excretion into urine of rats bearing Yoshida ascites sarcoma . Cancer Res. , 44 , 2387 – 2392 ( 1984. ). [PubMed] [Google Scholar]
  • 4. ) Kato , N. , Asanagi , M. , Moriyama , Y. , Fuse , H. , Rokutanda , M. and Fujimura , S.Studies on the liver thymidine kinase‐stimulating factor isolated from the cell‐free ascites fluid of the mouse bearing Ehrlich ascites tumor . Proc. Jpn. Cancer Assoc. 46th Annu. Meet. , 240 ( 1987. ) ( in Japanese ). [Google Scholar]
  • 5. ) Tzonou , A. , Spares , L. , Kalapothaki , V. , Zavitsanos , X. , Rebelakos , A. and Trichopoulos , D.α1Antitrypsin and survival in hepatocellular carcinoma . Br. J. Cancer , 61 , 72 – 73 ( 1990. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 6. ) Bhattacharyya , J. and Chaudhuri , L.α1‐Antitrypsin: a possible tool for diagnosis of cervical cancer . Biochem. Med. Metab. Biol , 43 , 243 – 245 ( 1990. ). [DOI] [PubMed] [Google Scholar]
  • 7. ) Nishida , M. , Kasahara , K. , Kaneko , M. and Iwasaki , H.Establishment of a new human endometrial adenocar‐cinoma cell line, Ishikawa cells, containing estrogen and progesterone receptors . Acta Obstet. Gynecol. Jpn. , 37 , 1103 – 1111 ( 1985. ) ( in Japanese ). [PubMed] [Google Scholar]
  • 8. ) Motley , C. G. D. and Kingdon , H. S.Use of 3H‐thymidine for measurement of DNA synthesis in rat liver–a warning . Anal. Biochem. , 45 , 298 – 305 ( 1972. ). [DOI] [PubMed] [Google Scholar]
  • 9. ) Laskowski , M.Trypsinogen and trypsin . Methods Enzymol , 2 , 26 – 36 ( 1955. ). [Google Scholar]
  • 10. ) Shimada , H. , Mori , T. , Takada , A. , Takada , Y. , Noda , Y. , Takai , L , Kohda , H. and Nishimura , T.Use of chromogenic substrate S‐2251 for determination of plasminogen activator in rat ovaries . Thrombos. Haemostas. , 46 , 507 – 510 ( 1981. ). [PubMed] [Google Scholar]
  • 11. ) Laemmli , U. K.Cleavage of structural proteins during the assembly of the head of bacteriophage T4 . Nature , 227 , 189 – 198 ( 1970. ). [DOI] [PubMed] [Google Scholar]
  • 12. ) Ouchterlony , O.Diffusion‐in‐gel methods for immunological analysis . Prog. Allergy , 5 , 1 – 78 ( 1958. ). [PubMed] [Google Scholar]
  • 13. ) Lowry , O. H. , Rosebrough , N. J. , Farr , A. L. and Randall , R. J.Protein measurement with the Folin phenol reagent . J. Biol. Chem. , 193 , 265 – 275 ( 1951. ). [PubMed] [Google Scholar]
  • 14. ) Kato , N. , Moriyama , Y. and Fujimura , S.Studies on the liver thymidine kinase‐stimulating factor isolated from the cell‐free ascitic fluid of the mouse bearing Ehrlich ascites tumor (abstract) . Proc. Jpn. Biochem. Soc. , 57 , 1199 ( 1985. ) ( in Japanese ). [Google Scholar]
  • 15. ) Carrell , R. W. , Jeppsson , J‐O. , Laurell , C‐B. , Brennan , S. O. , Owen , M. C. , Vaughan , L. and Boswell , D. R.Structure and variation of human arantitrypsin . Nature , 298 , 329 – 333 ( 1982. ). [DOI] [PubMed] [Google Scholar]
  • 16. ) Long , G. L. , Chandra , T. , Woo , S. L. C. , Davie , E. W. and Kurachi , K.Complete sequence of the cDNA for human a1‐antitrypsin and the gene for the S variant . Biochemistry , 23 , 4828 – 4837 ( 1984. ). [DOI] [PubMed] [Google Scholar]
  • 17. ) Asanagi , M. , Tsukifuji , R. , Okamoto , S. , Rokutanda , M. , Kato , N. , Moriyama , Y. and Fujimura , S.Activities of thymidylate synthetase and thymidine kinase in the liver of the mouse bearing B‐16 melanoma or Ehrlich ascites tumor . Proc. Jpn. Cancer Assoc. 46th Annu. Meet. , 238 ( 1987. ) ( in Japanese ). [Google Scholar]
  • 18. ) Nukiwa , T. , Satoh , K. , Brantly , M. L. , Ogushi , F. , Fells , G. A. , Courtney , M. and Crystal , R. G.Identification of a second mutation in the protein‐coding sequence of the Z type alpha 1‐antitrypsin gene . J. Biol. Chem. , 261 , 15989 – 15994 ( 1986. ). [PubMed] [Google Scholar]
  • 19. ) Nukiwa , T. , Brantly , M. , Ogushi , F. , Fells , G. , Satoh , K. , Stier , L. , Courtney , M. and Crystal , R.G. Characterization of the M 1 (Ala213) type of α1‐antitrypsin, a newly recognized, common “normal”α1‐antitrypsin haplotype . Biochemistry , 26 , 5259 – 5267 ( 1987. ). [DOI] [PubMed] [Google Scholar]
  • 20. ) Nukiwa , T. , Takahashi , H. , Brantly , M. , Courtney , M. and Crystal , R. G.α1Antitrypsin NullGranite Falls, a non‐expressing α1‐antitrypsin gene associated with a frameshift to stop mutation in a coding exon . J. Biol Chem. , 262 , 11999 – 12004 ( 1987. ). [PubMed] [Google Scholar]
  • 21. ) Jeppsson , J‐O. , Lilja , H. and Johansson , M.Isolation and characterization of two minor fractions of α1antitrypsin by high‐performance liquid chromatographic chromatofocusing . J. Chromatogr. , 327 , 173 – 177 ( 1985. ). [DOI] [PubMed] [Google Scholar]
  • 22. ) Vaughan , L. and Carrell , R.α1Antitrypsin isoforms: structural basis of microheterogeneity . Biochem. Int. , 2 , 461 – 467 ( 1981. ). [Google Scholar]
  • 23. ) Vaughan , L. , Lorier , M. A. and Carrell , R. W.α1Antitrypsin microheterogeneity: isolation and physiological significance of isoforms . Biochim. Biophys. Acta , 701 , 339 – 345 ( 1982. ). [DOI] [PubMed] [Google Scholar]
  • 24. ) Thompson , S. , Guthrie , D. and Turner , G. A.Fucosylated forms of alpha‐1‐antitrypsin that predict unresponsiveness to chemotherapy in ovarian cancer . Br. J. Cancer , 58 , 589 – 593 ( 1988. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 25. ) Chawla , R. K. , Rausch , D. J. , Miller , F. W. , Vogler , W. R. and Lawson , D. H.Abnormal profile of serum proteinase inhibitors in cancer patients . Cancer Res. , 44 , 2718 – 2723 ( 1984. ). [PubMed] [Google Scholar]
  • 26. ) Chawla , R. K. , Lawson , D. H. , Sarma , P. R. , Nixon , D. W. and Travis , J.Serum α‐1 proteinase inhibitor in advanced cancer: mass variants and functionally inert forms . Cancer Res. , 47 , 1179 – 1184 ( 1987. ). [PubMed] [Google Scholar]
  • 27. ) Ä stedt , B. , Svanberg , L. and Nilsson , I. M.Fibrin degradation products and ovarian tumours . Br. Med. J. , 4 , 458 – 459 ( 1971. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 28. ) Svanberg , L. and Ästedt , B.Coagulative and fibrinolytic properties of ascitic fluid associated with ovarian tumors . Cancer , 35 , 1382 – 1387 ( 1975. ). [DOI] [PubMed] [Google Scholar]
  • 29. ) Clemmensen , I. and Christensen , F.Inhibition of urokinase by complex formation with human α anti‐trypsin . Biochim. Biophys. Acta , 429 , 591 – 599 ( 1976. ). [DOI] [PubMed] [Google Scholar]
  • 30. ) Harada , N. , Shirasaka , T. and Fujii , S.DNA synthesis in tumor‐bearing rats: purification of liver thymidine kinase stimulating factor from Yoshida sarcoma . Gann , 71 , 173 – 180 ( 1980. ). [PubMed] [Google Scholar]
  • 31. ) Sasaki , K. , Imamura , K. and Tanaka , T.Mechanism of hepatic ornithine decarboxylase induction by the ornithine decarboxylase‐inducing factor isolated from tumor ascites fluid: determination of target cells for the factor in the liver . J. Biochem. , 94 , 949 – 959 ( 1983. ). [DOI] [PubMed] [Google Scholar]
  • 32. ) Kojima , Y. and Sakurada , T.Increase in alkaline phosphatase activity in the liver of mice bearing Ehrlich ascites tumor . Cancer Res. , 36 , 23 – 27 ( 1976. ). [PubMed] [Google Scholar]
  • 33. ) Duffy , M. JDo proteases play a role in cancer invasion and metastasis? Eur. J. Cancer Clin. Oncol , 23 , 583 – 589 ( 1987. ). [DOI] [PubMed] [Google Scholar]

Articles from Japanese Journal of Cancer Research : Gann are provided here courtesy of Wiley

RESOURCES