The purpose of this study was to investigate the cause of polycythemia occurring in hepatocellular carcinoma-bearing control male B6C3F, mice from 2-yr carcinogenicity studies. Erythrocyte counts and plasma levels of erythropoietin in mice with hepatocellular carcinomas were significantly increased compared with the values in non-tumor-bearing mice. Erythropoietin mRNA in 4 of 5 non-tumor-bearing mice was detected in the kidney, but no visible signals for hepatic erythropoietin mRNA in 5 of 5 non-tumor-bearing mice were detected by the reverse transcriptase competitive polymerase chain reaction method. Erythropoietin mRNA was expressed in neoplastic hepatocytes from 8 of 9 hepatocellular carcinoma-bearing mice, and this expression was accompanied by decreased expression of erythropoietin mRNA in the kidneys from these mice. The present findings show that polycythemia in hepatocellular carcinoma-bearing mice occurs secondary to excess synthesis of erythropoietin mRNA by neoplastic hepatocytes.
1. Bondurant MC and Koury MJ (1986). Anemia induces accumulation of erythropoietin mRNA in the kidney and liver. Mol. Cell Biol.. 6: 2731–2733.
2.
2. Bondurant MC, Koury MJ, and Semenza G (1991). Erythropoietin ontogeny and organ distribution in mice. Semin. Hematol.. 28: 20–27.
3.
3. Caro J, Zon. LI, Silver R, Miller O, and Erslev AJ (1983). Erytho-poetion in liver tissue extracts and liver purfusates from hypoxic rats. Am. J. Physiol.. 244: E431–E434.
4.
4. Castle WB (1985). The polycythemias. In: Hematology,4th ed., WS Becker (ed). MIT Press, Cambrigde, Massachusetts, pp. 305–321.
5.
5. Chandra M and Frith CH (1992). Spontaneous neoplasmas in B6C3F, mice. Toxicol. Lett.. 60: 91–98.
6.
6. DeGowin RL, Hofstra D, and Gurney CW (1962). A comparison of erythropoietin bioassays. Proc. Soc. Exp. Biol. Med.. 110: 48–51.
7.
7. Eckardt KU, Ratclitte PJ, Tan CC, Bauer C, and Kurtz A (1992). Age dependent expression of the erythropoietin gene in rat liver and kidney. J. Clin. Invest.. 89: 753–760.
8.
8. Fandrey J and Bunn FH (1993). In vivoand in vitroregulation of erythropoietin mRNA: Measurement by competitive polymerase chain reaction. Blood. 81: 617–623.
9.
9. Goldberg MA, Glass GA, Cunningham JM, and Bunn HF (1986). The regulated expression of erythropoietin by 2 human hepatoma cell lines. Proc. Natl. Acad. Sci. USA. 84: 7972–7976.
10.
10. Haseman JK, Huff JE, Roo GN, Arnold JE, Boorman GA, and McConnell EE (1985). Neoplasm observed in untreated and corn oil gavage control groups of F344/N rats and (C57BL/6N X C3H/ HeN)F1 (B6C3F1) mice. JNC175: 975–984.
11.
11. Herkens C, Wolff M, Fandrey J, Schuler F, and Jelkmann W (1990). Immuno-cytochemical demonstration of erythropoietin in hypoxic human hepatoma cultures. Histochemistry. 100: 303–309.
12.
12. Horinouchi A, Hayashi S, Ando T, and Nonoyama T (1996). Polycythemia in hepatocellular carcinoma-bearing B6C3F1 mice. J. Toxicol. Pathol.. 9: 233–240.
13.
13. Horinouchi A, Hayashi S, Ando T, and Nonoyama T (1997). Secondary polycythemia in male B6C3F1 mice with spontaneously occurring hepatocellular carcinomas. Toxicol. Pathol.25: 511–515
14.
14. Horinouchi A, Takami K, Sekiguchi M, and Ando T (1996). Erythropoietin messenger RNA expression in the kidneys of mice with acute anemia. J. Toxicol. Pathol.. 9: 51–56.
15.
15. Kan YW, McFadzean AJS, Todd D, and Tso SC (1961). Further observations on polycythemia in hepatocellular carcinoma. Blood. 18: 592–598.
16.
16. Maronport RR, Haseman JK, Boorman GA, Eustis SE, Rao GN, and Huff JE (1987). Liver lesions in B6C3F1 mice: The National Toxicology Program, experience and position. Arch. Toxicol. Suppl.. 10: 10–26.
17.
17. McFadzean AJS, Todd D, and Tsang KC (1958). Polycythemia in primary carcinoma of the liver. Blood. 13: 427–435.
18.
18. Muta H, Funakoshi A, Baba T, Ueki N, Wakasugi H, Kozuru M, and Jimi A (1994). Gene expression of erythropoietin in hepatocellular carcinoma. Intern. Med.. 33: 427–431.
19.
19. Nemi CJ (1986). Clinical and laboratory evaluation of anemias and polycythemia. In: Shalm's Veterinary Hematology, 4th ed., CJ Nemi (ed). Lea & Febiger, Philadelphia, pp. 572–576.
20.
20. Powell DJ, Friedman JM, Ouletter AJ, Krauter KS, and Darnell JE Jr (1984). Transcriptional and post-transcriptional control of specific messenger RNAs in adult and embryonic liver. J. Mol. Biol.. 179: 21–35.
21.
21. Sakisaka S, Watanabe M, Tateishi H, Harada M, Shakado S, Mi-mura Y, Gondo K, Yoshitake M, Noguchi K, Hino T, Nohno R, Majima Y, Hirai K, Sata M, Yoshida H, and Tanikawa K (1993). Erythropoietin production in hepatocellular carcinoma cells associated with polycythemia: Immunohistochemical evidence. Hepa-tology. 18: 1357–1362.
22.
22. Schemaker CB and Mitstock LD (1986). Murine erythropoietin gene: Cloning, expression, and human gene homology. Mol. Cell Biol.. 6: 849–858.
23.
23. Snedecor GW and Cochran WG (1980). Statistical Methods,7th ed.Iowa State University Press, Ames.
24.
24. Statistical Analysis Systems (1992). SAS Technical Report, SAS/ STAT Software,Release 6.07. Statistical Analysis Systems Institute, Cary, North Carolina.
25.
25. Tamano S, Hagiwara A, Shibata M, Kurata Y, Fukushima S, and Ito N (1988). Spontaneous tumors in aging (C57BL/6N X C3H/ HeN)F1 (B6C3F1) mice. Toxicol. Pathol.. 3: 321–326.
26.
26. Zanjani ED, Poster J, Burlington H, Mann L, and Wasserman LR (1977). Liver as the primary site of erythropoietin formation in the fetus. J. Lab. Clin. Med.. 89: 640–644.
