Expression of human metallothionein-III confers protection against serum-free exposure of stably transfected Chinese hamster ovary CHO-K1 cells

被引：5

作者：

Amoureux, MC ^{[1
]}

Wurch, T ^{[1
]}

Pauwels, PJ ^{[1
]}

机构：

[1] CTR RECH PIERRE FABRE, CELLULAR & MOLEC NEUROBIOL LAB, F-81106 CASTRES, FRANCE

来源：

NEUROSCIENCE LETTERS | 1995年 / 201卷 / 01期

关键词：

metallothionein-III; (cloned; human); Chinese hamster ovary (CHO-K1) cell line; permanent transfection; reverse transcription-polymerase chain reaction; cell survival; serum starvation; growth factor;

D O I：

10.1016/0304-3940(95)12134-P

中图分类号：

Q189 [神经科学];

学科分类号：

071006 ;

摘要：

The cloned human metallothionein (MT)-III coding region was permanently transfected in Chinese hamster ovary (CHO-K1) cells in order to investigate the growth regulatory effects of this brain-specific protein. Reverse transcription-polymerase chain reaction (RT-PCR) experiments demonstrated stable expression of MT-III mRNA in CHO-K1/MT-III cells. Whereas in the presence of serum no differences were observed between the cell growth of both CHO-K1/MT-III and CHO-K1/pcDNA3-plasmid transfected cells, cell survival was attenuated differently in serum-free medium. CHO-K1/MT-III cells were more resistant (40 +/- 8%) to serum deprivation than pcDNAS-plasmid transfected cells. Recovery of cell growth for both cell lines was obtained by supplementing serum (0.25%), although with a different growth rate; half-maximal 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT)-conversion was observed between 5.1-5.7 and 4.2-4.5 days for CHO-K1/pcDNA3 and CHO-K1/MT-III cells, respectively. These results suggest a protective role for cloned human MT-III, besides its reported inhibitory and stimulatory effects on cell survival.

引用

页码：61 / 64

页数：4

共 23 条

[1] MODULATION OF METALLOTHIONEIN-III MESSENGER-RNA CONTENT AND GROWTH-RATE OF RAT C6-GLIAL CELLS BY TRANSFECTION WITH HUMAN 5-HT1D RECEPTOR GENES [J].

AMOUREUX, MC ;

WURCH, T ;

PAUWELS, PJ .

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1995, 214 (02) :639-645

[2] NEUROPIL THREADS OCCUR IN DENDRITES OF TANGLE-BEARING NERVE-CELLS [J].

BRAAK, H ;

BRAAK, E .

NEUROPATHOLOGY AND APPLIED NEUROBIOLOGY, 1988, 14 (01) :39-44

[3] THE RELATIVE IMPORTANCE OF GLUTATHIONE AND METALLOTHIONEIN ON PROTECTION OF HEPATOTOXICITY OF MENADIONE IN RATS [J].

CHAN, HM ;

TABARROK, R ;

TAMURA, Y ;

CHERIAN, MG .

CHEMICO-BIOLOGICAL INTERACTIONS, 1992, 84 (02) :113-124

[4] DETECTION OF NGF-LIKE ACTIVITY IN HUMAN BRAIN-TISSUE - INCREASED LEVELS IN ALZHEIMERS-DISEASE [J].

CRUTCHER, KA ;

SCOTT, SA ;

LIANG, S ;

EVERSON, WV ;

WEINGARTNER, J .

JOURNAL OF NEUROSCIENCE, 1993, 13 (06) :2540-2550

[5] ENHANCED NEUROTROPHIC ACTIVITY IN ALZHEIMERS-DISEASE CORTEX IS NOT ASSOCIATED WITH DOWN-REGULATION OF METALLOTHIONEIN-III (GIF) [J].

ERICKSON, JC ;

SEWELL, AK ;

JENSEN, LT ;

WINGE, DR ;

PALMITER, RD .

BRAIN RESEARCH, 1994, 649 (1-2) :297-304

[6] MASSIVE SOMATODENDRITIC SPROUTING OF CORTICAL-NEURONS IN ALZHEIMERS-DISEASE [J].

IHARA, Y .

BRAIN RESEARCH, 1988, 459 (01) :138-144

[7] MEMORIES OF METALLOTHIONEIN [J].

KILLE, P ;

HEMMINGS, A ;

LUNNEY, EA .

BIOCHIMICA ET BIOPHYSICA ACTA-PROTEIN STRUCTURE AND MOLECULAR ENZYMOLOGY, 1994, 1205 (02) :151-161

[8] MOLECULAR-CLONING OF RAT GROWTH-INHIBITORY FACTOR CDNA AND THE EXPRESSION IN THE CENTRAL-NERVOUS-SYSTEM [J].

KOBAYASHI, H ;

UCHIDA, Y ;

IHARA, Y ;

NAKAJIMA, K ;

KOHSAKA, S ;

MIYATAKE, T ;

TSUJI, S .

MOLECULAR BRAIN RESEARCH, 1993, 19 (03) :188-194

[9] NEUROTROPHIC FACTORS - FROM MOLECULE TO MAN [J].

LINDSAY, RM ;

WIEGAND, SJ ;

ALTAR, CA ;

DISTEFANO, PS .

TRENDS IN NEUROSCIENCES, 1994, 17 (05) :182-190

[10] AN AUTOMATED COLORIMETRIC MICROASSAY FOR NEURONOTROPHIC FACTORS [J].

MANTHORPE, M ;

FAGNANI, R ;

SKAPER, SD ;

VARON, S .

DEVELOPMENTAL BRAIN RESEARCH, 1986, 25 (02) :191-198

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