Department of Biochemistry

1. Gene Expression during Stress Responses in Mammalian Cells(K.H.)
2. Senescence and Immortalization of Human Cells(S.G.)
3. Regulation of Patho-histogenesis by Gene Expression(H.H.)
4. Reactive Oxygen Species and Defense Mechanism(T.A.)

1. Gene Expression during Stress Responses in Mammalian cells:

   We are investigating the regulatory mechanism of the stress gene expression and the changes in stress proteins during responses against various stresses. Classical research using cultured cell system failed to detect the involvement of either neuroendocrinal factors or various cytokines in the regulation of stress protein gene expression. Recently, studies on the regulatory mechanism of heat shock gene expression by combination of heat treatment with several endogenous factors are in progress in this laboratory.

2. Senescence and Immortalization of Human Cells:

   Normal human cells have a limited in vitro life span, senesce, and finally perish at all. Among SV40-transformed human cells, however, immortalized cells could appear, although very rarely. So far, very little is known about the molecular mechanisms of immortalization which is an essential step toward cancer. The following two items are now under study in our laboratory.
   (I)regulation of telomere length
   (II)regulation of telomerase activity

3. Regulation of Patho-histogenesis by Gene Expression

   Liver cirrhosis, proliferative nephritis, tendon granulation etc. are known as incurable disease. These pathogenic histogenesis (from fibrosis to cancer) are regulated by the extracellular matrix proteins (Profilin, Fibronectin, Collagens, Woundhealin etc) and reguratory proteins (Kinase C, c-JUN etc.), therefore the investigation of fibrogenesis which originated in aberration of histogenesis have been investigating [idmc].

4. Reactive Oxygen Species and Defense Mechanism:

   We have investigated the defense mechanisms against reactive oxygen species generated by either cadmium or anti-tumor drugs in relation to glutathione, metallothionein and heat shock proteins. Recently, our attention is focuced on the signal transduction from the oxidative stress to nuclear gene expression of various proteins for defense mechanisms.
   We also investigate protective mechanisms of apoptosis caused by oxygen radicals.
   

Nakagawara A, Ikeda K, Higashi K, Sasazuki T (1990): Inversed correlation between N-myc amplification and catecholamine metabolism in children with advanced neuroblastoma.
Surgery 107: 43-49.

Noguchi S, Higashi K, & Kawamura M (1990):Assembly of the Alpha-subunit of Torpedo california Na+/K+-ATPase with its pre-existing Beta-subunit in Xenopus oocytes. Biochim Biophys Acta 1023:247-253.

Katoh T, Karasaki Y, Hirano H, Gotoh S, & Higashi K (1990): Translocation of protein kinase C to membrane induced by TNF does not cause the inhibition of EGF binding to human WISH cells.
Biochem Biophys Res Comm 168:690-695.

Ohmori H, Murakami T, Furutani A, Higashi K, Hirano H, Gotoh S, Kuroiwa A, Masui A, Nakamura T, & Amalric F (1990); Simultaneous activation of heat shock protein(hsp70) and nucleolin genes during in vivo and in vitro prereplicative stages of rat hepatocytes. Exp Cell Res 189:227-232.

Noguchi S, Higashi K & Kawamura M (1990); A possible role of the Beta-subunit of (Na,K)-ATPase in facilitating correct assembly of the Alpha-subunit into the membrane. J Biol Chem 265;15991-15995.

Katoh T, Higashi K, Inoue N, & Tanaka I (1990): Different responses of cytosolic and mitochondrial glutathione in rat liver after ethylene oxide exposure. Toxcol Lett 54:235-239.

(1991)

Yano O, Hirano H, Karasaki Y, Higashi K, Nakamura H, Akiya S, & Gotoh S (1991): Cloning and sequencing of viral integration site in human fibroblasts immortalized by simian virus 40 (1991): Cell Struct Funct 16:63-71.

Murakami T, Ohmori H, Gotoh S, Tsuda T, Ohya R, Akiya S, & Higashi K (1991): Down regulation of N-myc, heat-shock protein 70, and nucleolin during the differentiation of human neuroblastoma cells. J Biochem 110: 146-150.

Katoh T, Ohmori H, Murakami T, Karasaki Y, Higashi K, & Muramatsu M (1991): Induction of glutathione S-transferase and heat-shock proteins in rat liver after ethylene oxide exposure. Biochem Pharmacol 42: 1247-1254.

Murakami T, Ohmori H, Katoh T, & Higashi K (1991): Cadmium causes increases of N-myc and multidrug-resistance gene mRNA in neuroblastoma cells. J UOEH 13:271-278.

(1992)

Katoh T, Higashi K, Karasaki Y(1992): The mechanism of the action of IFN-Gamma and TPA on the modulation of epidermal growth factor receptors of human amnion cells. J UOEH 14:165-172.

Ohmori H, Abe T, Hirano H, Murakami T, Katoh T, Gotoh S, Kido M, Kuroiwa A, Nomura T, & Higashi K (1992): Comparison of Ki-ras gene mutation among simultaneously occurring multiple urethan-induced lung tumors in individual mice. Carcinogenesis 13:851-855.

Murakami T, Ohmori H, Katoh T, Nakamura H, Tsuda T, Ren Chan Shang, Akiya S, Nakagawara A & Higashi K (1992):Differences of L-myc polymorphic patterns of neuroblastoma in patients between less than one year and older ages. J Pediatr Surg 27:1442-1446.

Karasaki Y, Katoh T, Higashi K, & Gotoh S (1992): Different resposiveness of normal and simian virus 40-transformed human fibroblast cells to interferon-Gamma. J Interferon Res 12: 185-190.

Katoh T, & Higashi K (1992): Ethnic differences of the primary gene defect at the cytochrome P-450 2D6. J UOEH 14:205-209.

Hiranuma K, Hirata K, Abe T, Hirano T, Matsuno K, Hirano H, Suzuki K, & Higashi K (1992):
Induction of mitochondrial chaperonin, Hsp60, by cadmium in human hepatoma cells. Biochem Biophy Res Comm 194: 531-536,

(1994)

Satoh Y, Kashimura M, Kaneko S, Karasaki Y, Higashi K, & Gotoh S (1994): Cloning of cDNA with possible association with senescence and immortalization of human cells. Mut Res 316:25-36.

Abe T, Konishi T, Katoh T, Hirano H, Matsukuma K, Kashimura M,& Higashi K(1994): Induction of heat shock 70 mRNA by cadmium is mediated by glutathione suppressive and non-suppressive triggers, Biochim Biophys Acta 1201: 29-36.

Karasaki Y, Satoh Y, Ohji T, Tsukamoto S, Higashi K, Gotoh S & Mizusaki K(1994): The nucleotide sequence of a cDNA clone encoding ribosomal protein S17 of Ehrlich ascites carcinoma cells. Biochem Genetics 32:409-414.

(1995)

Abe T, Konishi T, Hirano T, Kasai H, Shimizu K, Kashimura M, & Higashi K (1995): Possible correlation between DNA damage induced by hydrogen peroxide and translocation of heat shock 70 protein into the nucleus Biochem Biophys Res Comm 206: 548-555,

Kaneko S, Satoh Y, Ikemura K, Konishi T, Ohji T, Karasaki Y, Higashi K, & Gotoh S (1995):
Alterations of expression of the cytoskeleton after immortalization of human fibroblasts, Cell Struct Funct 20: 107-115.

Konishi T, Karasaki Y, Nomoto M, Ohmori H, Shibata K, Abe T, Shimizu K, Itoh H, & Higashi K (1995): Induction of heat shock protein 70 and nucleolin and their intracellular distribution during early stage of liver regeneration. J Biochem 117:1170-1177.

Ohji T, Urano H, Shirahata A, Yamagishi M, Higashi K, Gotoh S, & Karasaki Y (1995):
Transforming growth factor Beta1 and Beta2 induce down modulation of thrombomodulin in human umbilical vein endothelial cells. Thrombosis and Haemostasis 73:812-818.

Katoh T, Kaneko S, Ohya R, Higashi K & Ikemura K (1995): Glutathione-S-transferase M1 gene polymorphism in patients with oral squamous carcinoma in relation to cigarette-smoking.
Asian J Oral Maxillofacial Surg 7: 13-17,

Konishi T, Nomoto M, Shimizu K, Abe T, Itoh H, Friedrich H, Gunther E & Higashi K (1995):
Dominant role of the second heat shock element for the expression of HSP70-1 gene in rat liver after whole body hyperthermia. J Biochem 118:1021-l029.

1. Assistant Professor Hideyasu Hirano M. D. & Ph. D.

2. Key word : patho-histological manipulation , DNA binding protein, fibrosis, fibronectin, profilin, gene expression programming

3. Research Achievement
   Hirano, H., Yamada, Y., . Yamada, K. M. et al. (1983) Isolation of Genomic DNA Clones Spanning the Entire Fibronectin Gene. Proc. Natl. Acad. Sci. USA 80: 46-50

   Tyagi, S. J., Hirano, H., & Pastan, I. (1983) Transcriptional Control of the Fibronectin Gene in Chick Embryo Fibroblasts Transformed by Rous Sarcoma Virus. J. Biol. Chem. 258: 5787-5793

   Nimmer, D., Bergtrom, G., Hirano, H. et al. (1987) Regulation of Plasma Fibronectin Biosynthesis by Glucocorticoids in Chick Hepatocyte Culture. J. Biol. Chem. 262: 10369-10375

   Utsumi, K., . Hirano, H., Fujiwara, H. et al. (1991) Adhesion of immature thymocytes to thymic stromal cells through fibronectin molecules and its significance for the induction of thymocyte differentiation. Proc. Natl. Acad. Sci. USA 88: 5685-5689

   Shibata, K., Mita, T., . Hirano, H. et al. (1993) An attempt to substitute the cell binding domain of human fibronectin in Ramuda-phage J protein: Computer design and expression Biochimie. 75: 459-465

   Nakamura, H., Hirata, K., . Hirano, H. et al. (1994) Increase of Hepatic mRNAS of Profilin, Actin and Extracellular Matrix Proteins after Carbon Tetrachloride Treatment and Partial Hepatectomy in Rats Biochem. Biophys. Res. Comm. 198: 568-873

   Hiranuma, K., Suzuki, K., . Hirano, H. et al. (1996): Extracellular matrices in peritendinous connective tissue after surgical injuly to the chicken flexor tendon. Arch Orthop. Trauma Surgery in press

   Hirano, H., Hirano, T., Nakano, M. et al (1996) A Novel Approach for Obataining acDNA Clone that Codes Functional Protein by Applying the Fuzzy Set Cloning Theory, Biomedical Fuzzy and Human Sciens 2 19-14

   Tamura, M., Tanaka, H., Hirano, T. et al, (1996) Enhanced glomerular profilin gene and protein expression in experimental mesangial proliferative glomerulonephritis. Biochem Biophys Res Commun. 222 683-687

   Hirano, H., Hirano, T., Hirata, K. et al (1996) Experimental liver fibrosis induced inrats receiving high doses of alcohol and alternating between regular and vitamin-depleted diets. Experientia 1996 52 710-715

Social Activity

Patho-histologiical regulation by gene expressions