JPWO2007102572A1 - Cell aging inhibitor - Google Patents

Abstract

SUMMARY A novel cell aging inhibitor capable of effectively and fundamentally suppressing cell aging has been disclosed. The cell aging inhibitor contains, as an active ingredient, a substance that inhibits the expression of a gene encoding G protein γ subunit 11 or a substance that inhibits the cell aging action of G protein γ subunit 11. According to the cell aging inhibitor, since induction of cell aging due to oxidative stress can be prevented, various applications based on cell rejuvenation are possible. In particular, it can be applied to the development of cosmetics, health foods, and pharmaceuticals that are effective in preventing aging.

Description

  The present invention relates to a cell aging inhibitor that suppresses cellular aging.

  Normal human cells lose their ability to divide and age (replicative senescence). Senescent cells induce specific genes through specific morphological and physiological changes. Moreover, normal cells show the same phenomenon as above by various treatments (premature senescence). These two phenomena are essentially equivalent. It is becoming clear that cellular aging causes a decrease in organ / tissue function and is a direct cause of adult disease and human aging. Experimentally, cell senescence can be induced by various means, but there is no doubt that the main cause in the body is the accumulation of cell damage due to active oxygen. Traditionally, reactive oxygen has been thought to damage many biological materials, causing stress, activating signal transduction pathways (MAP kinase cascade) and inducing cellular senescence. However, it is completely unknown why reactive oxygen effectively induces cellular senescence.

  Regarding the extension of the mitotic life of cells, administration of antioxidants, suppression of MAP kinase activity, destruction of cell cycle regulator p21, etc. are known. However, these known techniques do not mention the mechanism of action of oxidative stress, which is a major factor of aging.

Hayflick, L., Moorehead P.S.The serial cultivation of human diploid cell strains.Exp cell Res 25, 585-621 (1961). Herbig, U. & Sedivy, J. M. Regulation of growth arrest in senescence: Telomere damage is not the end of the story.Mech Ageing Dev (2005). Lin, A. W. et al. Premature senescence involving p53 and p16 is activated in response to constitutive MEK / MAPK mitogenic signaling.Genes Dev 12, 3008-19 (1998). Wang, W. et al. Sequential activation of the MEK-extracellular signal-regulated kinase and MKK3 / 6-p38 mitogen-activated protein kinase pathways mediates oncogenic ras-induced premature senescence. Mol Cell Biol 22, 3389-403 (2002). Sumikawa, E., Matsumoto, Y., Sakemura, R., Fujii, M. & Ayusawa, D. Prolonged unbalanced growth induces cellular senescence markers linked with mechano transduction in normal and tumor cells.Biochem Biophys Res Commun 335, 558-65 (2005). Jakab, M. et al. Mechanisms sensing and modulating signals arising from cell swelling.Cell Physiol Biochem 12, 235-58 (2002). Samarakoon, R. & Higgins, P. J. MEK / ERK pathway mediates cell-shape-dependent plasminogen activator inhibitor type 1 gene expression upon drug-induced disruption of the microfilament and microtubule networks.J Cell Sci 115, 3093-103 (2002). Harman, D. Prolongation of the normal life span by radiation protection chemicals.J Gerontol 12, 257-63 (1957). Suzuki, T. et al. Induction of senescence-associated genes by 5-bromodeoxyuridine in HeLa cells.Exp Gerontol 36, 465-74 (2001). Minagawa, S., Fujii, M., Scherer, S. W. & Ayusawa, D. Functional and chromosomal clustering of genes responsive to 5-bromodeoxyuridine in human cells.Exp Gerontol in press (2004). Michishita, E. et al. 5-Bromodeoxyuridine induces senescence-like phenomena in mammalian cells regardless of cell type or species.J Biochem (Tokyo) 126, 1052-9 (1999). Gilman, A. G. G proteins: transducers of receptor-generated signals. Annu Rev Biochem 56, 615-49 (1987). Simon, M. I., Strathmann, M. P. & Gautam, N. Diversity of G proteins in signal transduction. Science 252, 802-8 (1991). Birnbaumer, L. Receptor-to-effector signaling through G proteins: roles for beta gamma dimers as well as alpha subunits.Cell 71, 1069-72 (1992). Wickman, K. D. et al. Recombinant G-protein beta gamma-subunits activate the muscarinic-gated atrial potassium channel.Nature 368, 255-7 (1994). Clapham, D. E. & Neer, E. J. New roles for G-protein beta gamma-dimers in transmembrane signaling.Nature 365, 403-6 (1993). Pitcher, J. A. et al. Role of beta gamma subunits of G proteins in targeting the beta-adrenergic receptor kinase to membrane-bound receptors. Science 257, 1264-7 (1992). Haga, K. & Haga, T. Activation by G protein beta gamma subunits of agonist- or light-dependent phosphorylation of muscarinic acetylcholine receptors and rhodopsin.J Biol Chem 267, 2222-7 (1992). Balcueva, E. A. et al. Human G protein gamma (11) and gamma (14) subtypes define a new functional subclass.Exp Cell Res 257, 310-9 (2000). Ray, K., Kunsch, C., Bonner, LM & Robishaw, JD Isolation of cDNA clones encoding eight different human G protein gamma subunits, including three novel forms designated the gamma 4, gamma 10, and gamma 11 subunits.J Biol Chem 270, 21765-71 (1995). Lim, WK, Myung, CS, Garrison, JC & Neubig, RR Receptor-G protein gamma specificity: gamma11 shows unique potency for A (1) adenosine and 5-HT (1A) receptors. Biochemistry 40, 10532-41 (2001) . Ide, A., Fujii, M., Nakababashi, K. & Ayusawa, D. Suppression of senescence in normal human fibroblasts by introduction of dominant-negative p53 mutants or human papilloma virus type 16 E6 protein.Biosci Biotechnol Biochem 62, 1458- 60 (1998). Brown, J. P., Wei, W. & Sedivy, J. M. Bypass of senescence after disruption of p21CIP1 / WAF1 gene in normal diploid human fibroblasts. Science 277, 831-4 (1997). Wei, W., Herbig, U., Wei, S., Dutriaux, A. & Sedivy, J. M. Loss of retinoblastoma but not p16 function allows bypass of replicative senescence in human fibroblasts.EMBO Rep 4, 1061-6 (2003). Chen, Q. M. et al. Molecular analysis of H2O2-induced senescent-like growth arrest in normal human fibroblasts: p53 and Rb control G1 arrest but not cell replication.Biochem J 332 (Pt 1), 43-50 (1998). DeYulia, GJ, Jr., Carcamo, JM, Borquez-Ojeda, O., Shelton, CC & Golde, DW Hydrogen peroxide generated extracellularly by receptor-ligand interaction facilitates cell signaling.Proc Natl Acad Sci USA 102, 5044-9 (2005 ). Mukhin, YV et al. 5-Hydroxytryptamine1A receptor / Gibetagamma stimulates mitogen-activated protein kinase via NAD (P) H oxidase and reactive oxygen species upstream of src in chinese hamster ovary fibroblasts.Biochem J 347 Pt 1, 61-7 (2000) . Matsukawa, J., Matsuzawa, A., Takeda, K. & Ichijo, H. The ASK1-MAP kinase cascades in mammalian stress response.J Biochem (Tokyo) 136, 261-5 (2004).

  An object of the present invention is to provide a novel cell aging inhibitor capable of effectively and fundamentally suppressing cell aging.

  As a result of searching various genes by gene array analysis, the present inventors have found various genes whose expression is increased in aging cells. Of these, attention was paid to the gene of γ subunit 11 of G protein. And (1) cell senescence is induced immediately when the G protein γ subunit 11 gene is overexpressed in normal human fibroblasts, and (2) the antisense cDNA for the G protein γ subunit 11 gene (3) Experimentally confirmed that the G protein γ subunit 11 gene is induced very rapidly by an aging inducer such as hydrogen peroxide. Thus, the present inventors have found that cell senescence can be suppressed by inhibiting G protein γ subunit 11 or its gene.

  That is, the present invention provides a cell aging inhibitor containing, as an active ingredient, a substance that inhibits the expression of a gene encoding G protein γ subunit 11 or a substance that inhibits the cellular aging action of G protein γ subunit 11. . The present invention also relates to the use of a substance that inhibits the expression of a gene encoding G protein γ subunit 11 or a substance that inhibits the cellular aging action of G protein γ subunit 11 for the production of a cell aging inhibitor. provide. Furthermore, the present invention relates to cell aging comprising administering to an individual an effective amount of a substance that inhibits the expression of a gene encoding G protein γ subunit 11 or a substance that inhibits the cellular aging action of G protein γ subunit 11. Provide a suppression method.

  According to the present invention, a novel cell aging inhibitor capable of effectively and fundamentally suppressing cell aging has been provided. According to the cell aging inhibitor of the present invention, induction of cell aging due to oxidative stress can be prevented, and thus various applications based on cell rejuvenation are possible. In particular, it can be applied to the development of cosmetics, health foods, and pharmaceuticals that are effective in preventing aging.

It is a figure which shows the expression increase of GNG-11 gene in aged human normal fibroblast TIG-7. Total mRNA samples were prepared from young human normal fibroblasts TIG-7 (32 PDL), resting cells (5 days serum starved), and senescent cells (75 PDL), and GNG-11 mRNA levels were quantified by Northern blot analysis. It is a figure which shows the expression induction of the GNG-11 gene by hydrogen peroxide. Hydrogen peroxide was added to the culture solution of young human normal fibroblast TIG-7 (42PDL) to prepare a total mRNA sample. Changes in GNG-11 mRNA levels were quantified by Northern blot analysis. It is a figure which shows the expression induction of a cell aging marker gene. A GNG-11 expression vector was introduced into young human normal fibroblasts (32PDL), and Northern blot analysis was performed on all mRNA samples prepared after 3 days of culture. It is a figure which shows the prolongation aging of the division life by introduction | transduction of antisense cDNA. An antisense GNG-11 cDNA expression vector was introduced into the immediately preceding human normal fibroblast TIG-7 (PDL62), and the gene-transferred cells were selected with a G418 selection medium. Colonies were stained and the number was counted. It is a figure which shows the change of the division life by the introduction | transduction of a GNG-11 expression plasmid. Sense GNG-11 cDNA or antisense GNG-11 cDNA expression vector was introduced into young human normal fibroblast TIG-7 (PDL32, arrow), and the gene-transferred cells were selected with G418 selection medium. Colonies were isolated and their mitotic life was measured.

  The G protein γ subunit 11 (hereinafter sometimes abbreviated as “GNG11”) belongs to the γ subunit family of G proteins (ten are known). G protein is composed of α, β and γ subunits. When a ligand binds to a cell membrane receptor, the G protein is activated and dissociates into α, βγ, and γ subunits. It was shown that α subunit, βγ, and γ subunit bound to GTP regulate the activity of many enzymes and ionization channels. Unlike other subunits, GNG11 is modified by the farnesyl group and has features such as inability to interact with β2 subunits. GNG11 is expressed in all tissues, but its function is completely unknown. The Ras protein, which has the potential to function as a shunt for signaling transmission, has recently been increasingly involved in cellular senescence. The base sequence of GNG11 gene and the amino acid sequence of GNG11 encoded by it are known. For example, the base sequence of cDNA of human GNG11 gene and the amino acid sequence of GNG11 encoded by it are described in GenBank Accession No. NM_004126. (SEQ ID NOS: 1 and 2 in the sequence listing).

  As specifically described in the Examples below, (1) cell senescence is induced immediately when the GNG11 gene is overexpressed in normal human fibroblasts, and (2) the gene is detected by an antisense cDNA against the GNG11 gene. It has been experimentally confirmed that (3) the GNG11 gene is induced very rapidly by an aging inducer such as hydrogen peroxide. These experimental facts indicate that GNG11 is a gene that promotes cell senescence, and cell senescence can be suppressed by inhibiting the action of GNG11 or expression of the GNG11 gene. In particular, from the findings of (2) above, it has been clarified that a substance that inhibits the expression of the GNG11 gene or a substance that inhibits the cell aging action of GNG11 has a cell aging inhibitory action.

  It should be noted that the degree of cell aging is determined by observation of cell morphology (flattening and hypertrophy occurs when cells age) and β-galactosidase staining image known as an aging marker (β-galactosidase staining image as aging progresses). Can be examined by observing

  The cell aging inhibitor of the present invention contains a substance that inhibits the expression of the GNG11 gene or a substance that inhibits the cell aging action of GNG11 as an active ingredient. Examples of substances that inhibit the expression of the GNG11 gene include antisense nucleic acids, interfering RNAs, peptides, and plant components for the GNG11 gene.

  As described above, since the base sequence of the GNG11 gene is known (GenBank Accession No. NM_004126, SEQ ID NO: 1), an antisense nucleic acid can be easily prepared based on a conventional method. The antisense nucleic acid may be an antisense RNA, or an antisense cDNA that generates an antisense RNA by transcription in a cell. Antisense RNA for the GNG11 gene is RNA that hybridizes with mRNA transcribed from the GNG11 gene and inhibits translation of the mRNA. Antisense RNA is most preferably completely complementary to GNG11 mRNA, but if there is a degree of complementarity that allows it to hybridize with mRNA in the cell, one with a slight mismatch may be used. it can. In this case, the identity of the base sequence of the antisense RNA to the complementary strand of the mRNA of GNG11 is preferably 90% or more, and more preferably 95% or more. Here, the identity of the base sequence is determined by aligning the sequences so that the number of matching bases is the largest, and the number of matching bases is the total number of bases (if the total number of bases is different). The number divided by the number of shorter bases) is expressed as a percentage, and can be easily calculated by known software such as BLAST. The size of the antisense RNA preferably has a full length of GNG11 cDNA or 70% or more of the full length, but can be used if it can specifically hybridize with mRNA of GNG11, preferably 18 bases or more. Is possible. Since the coding region of the base sequence shown in SEQ ID NO: 1 is 352 nt to 573 nt, an antisense RNA that hybridizes to a region including the entire length of the coding region or a region within the coding region is preferable. Instead of the antisense RNA, a recombinant vector in which an antisense cDNA is incorporated and the antisense cDNA is transcribed in the cell to produce the antisense RNA can also be used (see Examples below). Since expression vectors for mammals are well known and various types are commercially available, commercially available expression vectors can be used. Preferably, various plasmid vectors and virus vectors used as vectors for gene therapy and gene vaccines can be used. Recombinant vectors obtained by incorporating DNA that produces the above-described antisense RNA against the GNG11 gene by transcription into these commercially available expression vectors can be preferably used as the active ingredient of the cell aging inhibitor of the present invention.

  Since the base sequence of the GNG11 gene is known, the expression of the GNG11 gene can also be inhibited by RNA interference method (RNAi). RNAi is a technique that utilizes the action of a double-stranded RNA having the same base sequence as a partial region of a target RNA that cleaves the partial region of the target RNA. In human cells, it is known that double-stranded short RNA (siRNA) having 21 to 23 bases cleaves RNA having the same base sequence as one strand of the siRNA. Such siRNA can be used to inhibit the expression of the GNG11 gene. In this case, the siRNA may be a double-stranded RNA having the same base sequence as an arbitrary partial region of the cDNA of GNG11 (SEQ ID NO: 1), but the mRNA translation can be reliably inhibited by cleavage of the mRNA. Among the base sequences shown in SEQ ID NO: 1, an interfering RNA having the same base sequence as the region in the coding region of 352nt to 573nt is preferable.

  Since it became clear that cell aging can be suppressed by inhibiting the expression of the GNG11 gene, a substance that inhibits the cell aging action of GNG11 itself can of course be used as an active ingredient of the cell aging inhibitor of the present invention. As such a substance, an antibody against GNG11 can be mentioned. The antibody may be a monoclonal antibody or a polyclonal antibody, but if it is a polyclonal antibody, antibodies against all exposed epitopes of GNG11 are included, so that the cellular senescence action of GNG11 can be reliably inhibited. Moreover, since GNG11 specifically binds to other subunits of the G protein, the binding domain of the α or β subunit of the G protein can be used in the same manner as the antibody. When GNG11 binds to the binding domain of the α or β subunit of G protein, it can no longer bind to the α or β subunit of G protein, so that it cannot express physiological activity and cell aging is also inhibited. It is thought.

  The cell aging inhibitor of the present invention can be used as a pharmaceutical and a component of cosmetics and health foods. More specifically, examples of the drug include drugs for diseases such as senile skin diseases, allergies, atopy, arteriosclerosis, and hypertension. Moreover, it can also be included in cosmetics as a skin cell aging inhibitory component. Furthermore, in order to prevent aging of the whole body, it can also be used as a component of food and drink.

  When the active ingredient is antisense RNA or interfering RNA for GNG11 gene, a solution in which antisense RNA or interfering RNA is dissolved in a medium such as physiological buffer saline can be directly injected into a tissue where suppression of cell aging is desired. . It can also be introduced into tissues by electric pulse method or ultrasonic method. The dose in this case is appropriately selected according to the symptoms and the like, but usually it may be about 1 pmol to 1 nmol of antisense RNA or interfering RNA per 1 g of the target tissue. In addition, in the case of a recombinant vector incorporating an antisense cDNA, a solution obtained by dissolving the recombinant vector in a medium such as physiological buffer saline is directly injected into a tissue, muscle, or subcutaneous where cell aging is desired to be suppressed. be able to. The dose in this case is appropriately selected according to the symptoms and the like, but usually it may be about 1 pmol to 1 nmol of recombinant vector per 1 g of the target tissue. Further, when the active ingredient is an anti-GNG11 antibody, it is preferable to directly inject a solution dissolved in a medium such as physiological buffer saline into a tissue in which suppression of cell aging is desired. The dose in this case is appropriately selected according to the symptoms and the like, but usually it may be about 1 μg to 100 μg of antibody per 1 g of the target tissue. Active ingredients such as antisense RNA, recombinant vector incorporating antisense cDNA, and anti-GNG11 antibody can be administered systemically by parenteral or oral administration such as intravenous administration, intramuscular administration, and subcutaneous administration. It is. The dose in the case of systemic administration is appropriately selected according to the symptoms and the like, but usually the dose per 1 g body weight is about 1/1000 to the same dose as in the case of direct injection described above. The amount is sufficient. In the case of antisense RNA or antibody, it can be applied to the skin as a cosmetic ingredient. In this case, the content in the cosmetic is appropriately selected, but may be about 1 μg to 1 mg per 1 g of the cosmetic preparation.

  When used as a medicine or cosmetic, the cell aging inhibitor of the present invention can be formulated according to a conventional method. In the case of a medicine, the active ingredient may be simply dissolved or suspended in a pharmaceutically acceptable medium such as physiological buffer saline, but additives commonly used for this may be added. In the case of cosmetics, the active ingredient of the cell aging inhibitor of the present invention may be simply dissolved or suspended in the cosmetic composition applied to the skin. An electric pulse method or an ultrasonic method can be used in combination. If the active ingredient of the cell aging inhibitor is a nucleic acid such as a recombinant vector incorporating antisense RNA, interfering RNA, or antisense cDNA, gene transfer is performed to increase the uptake of the nucleic acid into the cell. It is preferable to add lipids for use. This makes it easier for the nucleic acid to pass through the lipid bilayer constituting the cell membrane. Various lipids for gene introduction are commercially available. For example, Transfectin (trade name, manufactured by Bio-Rad), Lipofectamine (trade name, manufactured by Invitrogen), and the like can be used. The addition amount of these lipids for gene transfer is not particularly limited, but is usually about 1 μl to 100 μl based on the weight of the nucleic acid to be introduced (1 μg).

  Hereinafter, the present invention will be described more specifically based on examples. However, the present invention is not limited to the following examples.

1. Materials and methods
1) Cell culture Normal human fibroblast TIG-7 (available from Human Science Research Resource Bank) was used in this study. The cells were cultured in a plastic dish by adding 10% fetal calf serum to Dulbecco's modified Eagle medium under 37 ° C, 5% carbon dioxide, and 95% humidity. In this experiment, TIG-7 cells having a population doubling level (PDL) of 32 were used as young cells. TIG-7 cells cease dividing near PDL72 and age.

2) Transfection
GNG-11 cDNA (Genbank NM_004126) is amplified from total RNA of TIG-7 cells using a primer pair (5'-tggacccagtctcaaacttaac-3 '(SEQ ID NO: 3), 5'-cccaagacaaaactttatttgaa-3' (SEQ ID NO: 4)) did. cDNA (812 bp) was incorporated into pGEM-T easy vector (commercially available from Promega), and after sequencing, it was cloned into pCDNA 3.1 (-) (commercially available from Invitrogen) in the sense and antisense orientations. The plasmid (10 μg) was introduced into the cells using Transfectin (30 μL) (trade name, commercially available from Bio-Rad). Since the vector contains a neomycin resistance gene, stable transfectants were selected on media containing G418 (400 μg / ml) (commercially available from AG Scientific, Inc.).

3) Northern blot analysis Total RNA samples were prepared from cells using an extraction kit. Samples (15 μg per lane) were electrophoresed on a 1% formaldehyde-agarose gel and transferred to a nylon membrane (Hybond-N, Amersham). The membrane was hybridized with hnRNPhnRNP (heterogeneous nuclear ribonucleo-protein) C1 / C2 or GAPDH cDNA labeled with 32P. Subsequently, it was washed twice with a 0.1XSSC solution containing 0.1% SDS and a 2X SSC solution containing 0.1% SDS (65 ° C., 30 minutes) to prepare an autoradiography (FUJI X-ray film).

4) DNA synthesis test
For the DNA synthesis ability test, BrdU labeling and detection kit II (RocheDiagnostics) were used. Cells were cultured to 50% saturation density on coverslips. Cells were labeled with BrdU for 30 minutes at 37 ° C and fixed with ethyl alcohol for 30 minutes at -20 ° C. The cells were reacted with BrdU antibody for 30 minutes at 37 ° C, then incubated with Alexa Fluor 546 for 2 hours at 37 ° C and observed with a fluorescence microscope.

5) Aging-specific β-galactosidase assay Cells were fixed with 2% formaldehyde / 0.2% glutaraldehyde and stained solution [1 mg / ml 5-bromo-4-chloro-3-indolyl-β-D-galactoside , 40 mM sodium citrate phosphate (pH 6.0), 5 mM potassium ferricyanide, 5 mM potassium ferrocyanide, 150 mM sodium chloride and 2 mM magnesium chloride] were added, and the mixture was kept at 35 ° C.

6) Colony formation test Seed cells in a dish. Colonies were formed. Colonies containing 50 cells or more were counted after staining with methylene blue.

2. result
1) GNG11 gene expression analysis
GNG11 gene expression was significantly induced in aged TIG-7 cells (FIG. 1) and significantly induced in resting cells (FIG. 1). It was remarkably induced in HeLa cells administered with BrdU. This fact suggests that GNG11 is involved in signal transmission of cellular senescence. Responsiveness to oxidative stress, which is the main cause of cell aging, was examined (FIG. 2). Young TIG-7 cells (PDL32) were treated with peroxygen peroxide under various conditions and examined for GNG11 gene expression. As a result, strong expression induction was observed at low and high concentrations. In addition, significant expression induction was observed within 2 hours after treatment. Therefore, it can be said that the GNG11 gene is an early response gene to oxidative stress.

2) Induction of cell aging by forced expression of GNG 11 gene A plasmid expressing GNG11 was introduced into young TIG-7 cells (PDL32), and the effect of overexpression of GNG11 on cell aging was analyzed. Clone G418 resistant cells stably incorporating the gene showed cell hypertrophy and flattening at the colony formation stage and stopped dividing. A strong staining image of aging-specific β-galactosidase (SA-βGal), which is a typical cell aging marker, was observed. Cell morphological changes and mitotic arrest were marked and faster than other cell senescence induction systems. For this reason, it was difficult to collect senescent cells for analysis of aging markers. Therefore, GNG11-expressing rasmid was introduced into the cells, and the cells were collected after 3 days without selecting a drug. Total RNA samples were prepared from these cells and Northern blot analysis was performed. As a result, induction of a cell aging marker (collagenase-1) was observed (FIG. 3).

3) Prevention of cellular senescence by suppressing GNG11 gene expression
A plasmid expressing GNG11 antisense RNA was introduced into TIG-7 cells (62 PDL) just before senescence, and the effect on cell senescence was examined. First, colony formation was observed. When GNG11 gene expression was suppressed, the cells formed normal colonies (FIG. 4). In cells expressing antisense RNA, disappearance of aging markers and increase in DNA synthesis ability (measured using BrdU labeling kit) were observed. On the other hand, after introducing the plasmid into young TIG-7 cells (32 PDL), G418 resistant clones were selected and their mitotic life was followed (FIG. 5). As a result, in the antisense RNA-expressing cells, prolongation of mitotic life was observed about 20 times. From the above, it was shown that if the expression of GNG11 is decreased, cell senescence is suppressed and life extension is observed.

Four. Discussion According to this study, the increase in GNG11 most rapidly induced cell senescence, and antisense RNA inhibited cell senescence. Activation of Ras and MAP kinase (ERK, p38, JNK is a major component) can be seen in the induction of cellular senescence. Therefore, an effector molecule located downstream of GNG11 is likely to be Ras or a MAP kinase activator located upstream of it. Oxidative stress is thought to activate MAP kinase and induce cellular senescence. Recently, it has been reported that when a membrane receptor works, it generates active oxygen, which activates G-protein. Since GNG11 responds to active oxygen, it indicates that GNG11 is the most important regulator of cellular senescence, and the molecular basis from oxidative stress to cellular senescence has been elucidated. Cell senescence can be controlled by using DNA, RNA, peptides, etc. derived from GNG11 and natural and artificial compounds targeting GNG11. This will lead to the commercialization of pharmaceuticals, cosmetics and health supplements for the purpose of beauty health.

The N-terminal region of the GNG11 subunit lacks commonality with other components, and the interaction between subunits is also specific, suggesting a new function of the G protein. GNG11 and GNG1 have high base homology, but the former is expressed in all tissues except the brain, while the latter is suggested to be involved in light transmission in the retina. G protein is dissociated by various stimuli, and GTP-binding α subunit controls the activity of downstream effector molecules. Recently, it has been shown that a robust βγ complex regulates downstream effector molecules (adenyl cyclase subunits II and IV, phospholipase A2, phospholipase C subtype, Ca ²⁺ channel, etc.).

Claims (6)

  A cell aging inhibitor comprising, as an active ingredient, a substance that inhibits the expression of a gene encoding G protein γ subunit 11 or a substance that inhibits the cell aging action of G protein γ subunit 11.   The cell aging inhibitor according to claim 1, comprising an antisense nucleic acid against the G protein γ subunit gene, a vector containing the antisense nucleic acid, or an interference RNA as an active ingredient.   Use of a substance that inhibits expression of a gene encoding G protein γ subunit 11 or a substance that inhibits cell aging action of G protein γ subunit 11 for the production of a cell aging inhibitor.   The use according to claim 3, wherein an antisense nucleic acid against the G protein γ subunit gene, a vector containing the antisense nucleic acid, or an interference RNA is used.   A method for inhibiting cell aging, comprising administering to an individual an effective amount of a substance that inhibits expression of a gene encoding G protein γ subunit 11 or a substance that inhibits cell aging action of G protein γ subunit 11.   The method according to claim 5, comprising administering an effective amount of an antisense nucleic acid against the G protein γ subunit gene, a vector containing the antisense nucleic acid, or an interfering RNA to an individual.

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