Purification and characterization of zinc-binding protein from the liver of the partially hepatectomized rat

Ohtake, Hideki; Koga, Mutuyosi

doi:10.1042/bj1830683

Cited by 48 publications

(18 citation statements)

References 42 publications

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“…This is indicated by dramatic differences in gene expression (21,52,53). For example, after partial hepatectomy MT was observed to accumulate in regenerating liver cells, the classic in vivo prototype of Go cells that are reentering S phase and mitosis (54,55).…”

Section: Discussionmentioning

confidence: 99%

Cell cycle regulation of metallothionein in human colonic cancer cells.

Nagel

Vallée

1995

Proc. Natl. Acad. Sci. U.S.A.

138

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Elevated levels of metallothionein (MT) found in rapidly growing tissues such as neonatal liver and various types of human tumors have suggested a role for MT in cell proliferation. To further explore this possibility we investigated the concentration of MT in human colonic cancer The present study takes a different approach to discern the role of MT in cell proliferation. It uses a sensitive anti-MT monoclonal antibody (mAb) that recognizes an epitope of rat and human MT other than the amino terminus (20). This mAb makes it possible to examine physiological steady-state-i.e., ng-levels of MT at different stages of cell proliferation without the need for pathophysiological metal concentrations. Furthermore, we have employed synchronized, continually dividing cells-i.e., human colonic cancer (HT-29) cells that are progressing from G1 to S, which differs biochemically from the Go/S progression (21). Synchronization was achieved with compactin, an inhibitor of 3-hydroxy-3-methylglutarylcoenzyme A reductase, which provides GI synchrony without affecting gene expression or protein biosynthesis (22,23). We find that the MT concentration oscillates during progression of HT-29 cells through the cell cycle, and this appears to have functional significance.MATERIALS AND METHODS Chemicals. Compactin (Mevastatin) and D,L-mevalonic acid were purchased from Fluka; the inactive lactone form of compactin was converted to its active form as described (22); [methyl-3H]thymidine (6.7 Ci/mmol; 1 Ci = 37 GBq) was from New England Nuclear. Cell culture products were from BioWhittaker. All other chemicals were purchased from Sigma.Cell Culture, Synchronization, and Assay of DNA Synthesis. HT-29 cells (HTB 38; American Type Culture Collection) were grown in 162-cm2 culture flasks as described (24) with human transferrin (10 ,ug/ml) (GIBCO/BRL) and using gentamycin sulfate (25 ug/ml) as antibiotic. For synchronization, confluent cells were trypsinized, dispersed by multiple pipetting, and inoculated into six-well plates at 1.5 x 104 cells per cm2. The medium was changed twice and after 48-56 h replaced with fresh medium containing 50 ,uM compactin. Cells were maintained under these conditions for 28 h. The medium was then removed, the cells were washed with Hanks' Abbreviations: mAb, monoclonal antibody; MT, metallothionein; FITC, fluorescein isothiocyanate. 579The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

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Section: Discussionmentioning

confidence: 99%

Cell cycle regulation of metallothionein in human colonic cancer cells.

Nagel

Vallée

1995

Proc. Natl. Acad. Sci. U.S.A.

138

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“…This result shows active proliferation of glial cells in the newborn mouse, and so in the fetal brain glial cells may be actively proliferating and re newed. Ohtake and Koga [10] believed that MT was important in the regulation of DNA synthesis through the synthesis or activity of enzymes required for nucleic acid. Nishimura et al [11] also demonstrated the possible involvement of MT in cell proliferation and differentiation.…”

Section: Discussionmentioning

confidence: 99%

Metallothionein in Developing Human Brain

Suzuki

Nakajima

Otaki³

et al. 1994

Biological Signals

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Localization of metallothionein (MT) in the developing human brain was investigated by immunohistochemical techniques. Fetal brain at 21 weeks showed no MT expression. In 35-week-old fetuses, glial cells in the gray matter showed MT expression in the nucleus and perinuclear cytoplasm, but glial cells in the white matter showed MT not only in the nucleus and perinuclear cytoplasm but also in glial processes. At 40 weeks, glial cells in gray and white matter expressed immuno-reactive MT in the nucleus, cytoplasm and glial processes. Blood vessels were positive for MT. In the infant brain, there were clear differences in glial cells between protoplasmic and fibrous astrocytes; their nuclei, cytoplasm and processes were positive for MT, but the MT-positive glial processes of protoplasmic astrocytes were fine. In the child, typical protoplasmic astrocytes in gray matter and fibrous astrocytes in white matter were observed and their nuclei, cytoplasm and glial processes were positive for MT. Pia mater and blood vessels expressed MT in infants and children.

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“…MT was found in the nuclei, cytoplasmic process and vascular feet of both protoplasmic and fibrous astrocytes. MT may be involved in detoxification of heavy metals and may also play a role in the storage, transport, absorption, excretion and homeostasis of heavy metals in the tissues (6 (18) considered that MT was important in the regulation of DNA synthesis through the synthesis or activity of enzymes required for nucleic acid. Nishimura et al (17) have also demonstrated a possible involvement of MT in cell proliferation and differentiation.…”

Section: Discussionmentioning

confidence: 99%