KR101362184B1 - Method and kit for prediction of chloasma - Google Patents

Abstract

The present invention relates to a method for predicting blemish formation and a diagnostic kit for predicting blemish formation, and specifically, to check the expression level of genes expressed differently between blemish tissue and normal skin tissue to diagnose the possibility of blemish formation on the skin. Probes that hybridize with a polynucleotide of a sequence, one or more polynucleotides or complementary polynucleotides thereof; Primer pairs of polynucleotides of said gene sequence; Or it relates to a stain formation prevention method and diagnostic kit comprising at least one monoclonal antibody having an amino acid sequence encoded by the gene as a variable region. According to the present invention, the possibility of blemish formation on the skin can be sensitively and quickly diagnosed by using 23 kinds of genes and 5 kinds of low expression genes that are specifically expressed at the blemish lesion site as blemish marker genes.

Description

Method and kit for prediction of chloasma {Method and kit for prediction of chloasma}

The present invention relates to a method for predicting spot formation and a diagnostic kit for predicting spot formation, and specifically, in order to diagnose the possibility of formation of spots in the skin by checking the expression levels of genes differently expressed between the spot tissue and normal skin tissue, the gene At least one polynucleotide or a complementary polynucleotide thereof as a probe that hybridizes with a polynucleotide of the sequence; A primer pair of a polynucleotide of the gene sequence; Or, it relates to a method for predicting spot formation and a diagnostic kit comprising one or more monoclonal antibodies having an amino acid sequence encoded by the gene as a variable region.

The melanin pigment produced by melanocytes of the body's skin is a phenolic polymer material in the form of a complex of black pigment and protein, and plays an important role in inhibiting skin damage caused by ultraviolet rays irradiated from the sun. The excessive synthesis and accumulation of melanin not only causes serious aesthetic problems such as melasma, freckles and senile lentigo in the skin, but also promotes skin aging and causes skin cancer (Reference: Hearing VJ et al., FASEB J., 5: 2902-2909, 1991).

Melasma is a symptom of hyperpigmentation that occurs mainly on skin exposed to the sun. It has a characteristic that is generally symmetrical, and rarely occurs in areas such as the eyelids or under the chin, which are relatively less exposed to sunlight. Therefore, the symptoms worsen in the summer when the sun is strong, but it becomes lighter in the winter. It is not significantly affected by gender, race, or age, but is mostly found in Asian or Latino women. The cause is still unclear, but it is known that it is mainly caused by exposure to sunlight, hormonal changes due to pregnancy, malnutrition, and endocrine system diseases.

Since it is difficult to completely remove a spot that has occurred once, it is important to prevent it. If you can check the skin prone to spots in advance and take intensive preventive measures, you will be able to prevent the occurrence of spots more efficiently. However, as described above, since the cause of the spot has not yet been clearly identified, it is difficult to predict the formation of spots in advance.

Accordingly, the present inventors have completed the present invention by finding genes whose expression increases or decreases in the melasma lesion area compared to the normal skin area, and reconfirms the level of expression in the melasma area with respect to the found genes.

Accordingly, an object of the present invention is to provide a method and kit for determining the possibility of skin spot formation by measuring the expression level of genes whose expression is specifically increased or decreased in a spot tissue. Another object of the present invention is to provide a method for screening a melasma formation inhibitor by confirming whether a test compound promotes or inhibits the action of a protein encoded from the gene.

1 is a photograph showing the mRNA expression levels of H19, PCOLCE2, WIF1, SOAT1, PDZK1, THRSP, TMEM56 and CYR61 at normal and lesion sites of melasma patients by RT-PCR.
2 is a graph showing the expression levels of H19, IGFBP6 and PDZK1 genes in normal and lesion sites of melasma patients by real time PCR.

In order to achieve the above object, the present invention is F7 (coagulation factor VII, serum prothrombin conversion accelerator), MGC34923 (hypothetical protein MGC34923), FLJ21963 (FLJ21963 protein), PRKAA1 (protein kinase, AMP-activated, alpha 1 catalytic subunit), ELOVL5 (ELOVL family member 5, elongation of long chain fatty acids), ACSM3 (acyl-CoA synthetase medium-chain family member 3), DGAT2L3 (diacylglycerol O-acyltransferase 2-like 3), GABRA4 (gamma-aminobutyric acid A receptor, alpha 4), TMEM56 (transmembrane protein 56), SLCO4C1 (solute carrier organic anion transporter family, member 4C1), THRSP (THYROID HORMONE-RESPONSIVE SPOT14), HAO2 (hydroxyacid oxidase 2 ), IGJ (immunoglobulin J polypeptide), ACSBG1 (acyl -CoA synthetase bubblegum family member 1), ACP6 (acid phosphatase 6, lysophosphatidic), SOAT1 (ACYL-CoA:CHOLESTEROL ACYLTRANSFERASE), ELOVL3 (elongation of very long chain fatty acids), C6orf188 (chromosome 6 open reading frame 1188), HSD3 (hydroxy-delta-5-steroid dehydrogenase, 3 beta- and steroid delta-isomerase 1), ML STD1 (male sterility domain containing 1), Cyr61 (CYSTEINE-RICH PROTEIN 61), DKFZP586H2123 (regeneration associated muscle protease), and PDZK1 (PDZ domain containing 1) one or more polynucleotides or fragments thereof selected from the group consisting of, the fragment is One or more polynucleotides comprising 10 or more contiguous nucleotides, or complementary polynucleotides thereof; And a solid support in which the polynucleotide is bonded to the surface, and if the amount of the transcript hybridized to the polynucleotide is greater than a normal value, it is diagnosed that the skin is easy to form spots. Provides.

In addition, as one or more polynucleotides or fragments thereof selected from the group consisting of PCOLCE2, GFPT2 (glutamine-fructose-6-phosphate transaminase 2), CCL21 (chemokine CC motif ligand 21), WIF1 (WNT inhibitory factor 1), and H19, the The fragment may comprise one or more polynucleotides comprising 10 or more contiguous nucleotides, or a complementary polynucleotide thereof; And a solid support in which the polynucleotide is bonded to the surface, and if the amount of the transcript hybridized to the polynucleotide is less than the normal value, it is diagnosed that the skin is easy to form spots. Kits are provided.

The present invention is composed of F7, MGC34923, FLJ21963, PRKAA1, ELOVL5, ACSM3, DGAT2L3, GABRA4, TMEM56, SLCO4C1, THRSP, HAO2, IGJ, ACSBG1, ACP6, SOAT1, ELOVL3, C6orf188, PDZKF1, C6orf188, HZPK1, and C6orf188, HZPK1Byr2 One or more polynucleotides comprising 18-22 contiguous nucleotides as fragments of polynucleotides selected from the group; And one or more polynucleotides comprising 18-22 contiguous nucleotides as a fragment of a polynucleotide complementary to the polynucleotide, and if the amount of DNA amplified by using the polynucleotides as a primer is greater than the normal value, it is considered that skin is easy to form spots. It provides a diagnostic kit for predicting the formation of blemishes on the skin, characterized in that for diagnosing.

In addition, the present invention is a fragment of a polynucleotide selected from the group consisting of PCOLCE2, GFPT2, CCL21, WIF1 and H19 at least one polynucleotide comprising 18-22 consecutive nucleotides; And one or more polynucleotides comprising 18-22 contiguous nucleotides as a fragment of a polynucleotide complementary to the polynucleotide, and if the amount of DNA amplified by using the polynucleotides as a primer is less than the normal value, it is easy to form spots. It provides a diagnostic kit for predicting the formation of spots on the skin, characterized in that the diagnosis is made.

The present invention is composed of F7, MGC34923, FLJ21963, PRKAA1, ELOVL5, ACSM3, DGAT2L3, GABRA4, TMEM56, SLCO4C1, THRSP, HAO2, IGJ, ACSBG1, ACP6, SOAT1, ELOVL3, C6orf188, PDZKF1, C6orf188, HZPK1, and C6orf188, HZPK1Byr2 It comprises at least one monoclonal antibody having an amino acid sequence encoded by a gene selected from the group as a variable region, and characterized in that it is diagnosed as skin with easy spot formation when the amount of antigen bound to the antibody is greater than the normal value. Provides a diagnostic kit for predicting the formation of blemishes on the skin.

In addition, the present invention includes one or more monoclonal antibodies having an amino acid sequence encoded by a gene selected from the group consisting of PCOLCE2, GFPT2, CCL21, WIF1 and H19 as a variable region, and the amount of antigen bound to the antibody is It provides a diagnostic kit for predicting the formation of spots on the skin, characterized in that it is diagnosed as the skin with easy spot formation when it is less than the normal value.

In addition, the present invention is a method for predicting the formation of blemishes in the epidermis. , C6orf188, HSD3B1, MLSTD1, Cyr61, DKFZP586H2123 and PDZK1 If the expression level of one or more genes selected from the group consisting of PDZK1 is greater than the normal value, it is determined that the skin is easy to form spots.

And, as a method of predicting the formation of spots on the skin, it is characterized in that it is determined that skin is easily spots formed when the expression level of one or more genes selected from the group consisting of PCOLCE2, GFPT2, CCL21, WIF1 and H19 in the epidermis is less than the normal value. It provides a method for predicting the formation of blemishes on the skin.

The present invention is a method for screening melasma formation inhibitors, F7, MGC34923, FLJ21963, PRKAA1, ELOVL5, ACSM3, DGAT2L3, GABRA4, TMEM56, SLCO4C1, THRSP, HAO2, IGJ, ACSBG1, ACP6, SOAT1, ELOVL3, ELOVL3 The test compound was added to a protein encoded by one or more genes selected from the group consisting of MLSTD1, Cyr61, DKFZP586H2123 and PDZK1, or a protein encoded by one or more genes selected from the group consisting of PCOLCE2, GFPT2, CCL21, WIF1 and H19. Combining; And determining whether the test compound promotes or inhibits the action of the protein.

Hereinafter, the present invention will be described in more detail.

In the present invention, 23 kinds of high-expression genes in the melasma region and five kinds of low-expression genes in the melasma region were discovered, and the fact that they are related to melasma has not been reported yet. Table 1 below lists genes that are specifically highly expressed in melasma tissue, and Table 2 lists genes that are specifically under-expressed. GB accession refers to genebank accession ID and Unigene refers to the ID of unigene DB, and discloses the sequence of each gene.

Genetic symbol GB_Accession UniGene Gene name F7 M13232 Hs.36989 coagulation factor VII (serum prothrombin conversion accelerator) MGC34923 BC027449 Hs.61232 hypothetical protein MGC34923 FLJ21963 BC009317 Hs.13222 FLJ21963 protein PRKAA1 BC037303 Hs.43322 protein kinase, AMP-activated, alpha 1 catalytic subunit ELOVL5 AF338241 Hs.343667 ELOVL family member 5, elongation of long chain fatty acids (FEN1/Elo2, SUR4/Elo3-like, yeast) ACSM3 AC004381 Hs.512678 acyl-CoA synthetase medium-chain family member 3 DGAT2L3 BC039181 diacylglycerol O-acyltransferase 2-like 3 GABRA4 BC035055 Hs.248112 gamma-aminobutyric acid (GABA) A receptor, alpha 4 TMEM56 AK056404 Hs.84522 transmembrane protein 56 SLCO4C1 BX647880 Hs.127648 solute carrier organic anion transporter family, member 4C1 THRSP BC031989 Hs.523924 thyroid hormone responsive (SPOT14 homolog, rat) HAO2 BC020863 Hs.236545 hydroxyacid oxidase 2 (long chain) IGJ BC038982 Hs.381568 immunoglobulin J polypeptide, linker protein for immunoglobulin alpha and mu polypeptides ACSBG1 BC009289 Hs.277543 acyl-CoA synthetase bubblegum family member 1 ACP6 BC009965 Hs.15871 acid phosphatase 6, lysophosphatidic SOAT1 BC028940 sterol O-acyltransferase (acyl-Coenzyme A: cholesterol acyltransferase) 1 ELOVL3 BC034344 Hs.302130 elongation of very long chain fatty acids (FEN1/Elo2, SUR4/Elo3, yeast)-like 3 C6orf188 BC032556 Hs.511968 chromosome 6 open reading frame 188 HSD3B1 M35493 Hs.364941 hydroxy-delta-5-steroid dehydrogenase, 3 beta- and steroid delta-isomerase 1 MLSTD1 BC022267 Hs.134497 male sterility domain containing 1 CYR61 AL832891 Hs.8867 cysteine-rich, angiogenic inducer, 61 DKFZP586H2123 AF370388 Hs.55044 regeneration associated muscle protease PDZK1 BC006518 Hs.15456 PDZ domain containing 1

Genetic symbol GB_Accession UniGene Gene name PCOLCE2 BC006265 Hs.8944 procollagen C-endopeptidase enhancer 2 GFPT2 BC000012 Hs.30332 glutamine-fructose-6-phosphate transaminase 2 CCL21 BC027918 Hs.57907 chemokine (C-C motif) ligand 21 WIF1 BC018037 Hs.284122 WNT inhibitory factor 1 H19 AK123560 H19, imprinted maternally expressed untranslated mRNA

Currently, what is known about these genes is as follows.

THRSP (THYROID HORMONE-RESPONSIVE SPOT14) is presumed to exhibit a function related to lipogenesis, but its exact function is not known, and its association with melasma is also unknown.

WIP1 (WILDTYPE p53-INDUCED PHOSPHATASE 1) is a protein that arrests the cell cycle by inactivating CHEK1 protein in a p53-dependent cell cycle, and there have been no reports of melasma.

SOAT1 (ACYL-CoA:CHOLESTEROL ACYLTRANSFERASE) is known to have a function related to the binding of acyl CoA to cholesterol.

Cyr61 (CYSTEINE-RICH PROTEIN 61) is known to promote cell proliferation, chemotaxis, angiogenesis, and cell adhesion. ), and is known to increase the expression of angiogenesis-related genes.

F7 (coagulation factor VII, serum prothrombin conversion accelerator) is a cofactor involved in blood coagulation.

PRKAA1 (protein kinase, AMP-activated, alpha 1 catalytic subunit) is a factor that regulates cholesterol synthesis and fatty acid synthesis through phospholyation, and is known to be involved in recognizing the level of ATP in cells. Nothing happened.

DGAT2L3 (diacylglycerol O-acyltransferase 2-like 3) is a type of diacylglycerol acyltransferase family, which is expected to play an important role in lipid metabolism in skin tissue. none.

GABRA4 (gamma-aminobutyric acid (GABA) A receptor, alpha 4) is a receptor for GABA, a representative inhibitory neurotransmitter in the nervous system.

HAO2 (hydroxyacid oxidase 2) is known to be an enzyme responsible for the oxidation of hydroxy acid in the peroxisome in the liver and kidney.

IGJ (immunoglobulin J polypeptide) is a linker protein that connects immunoglobulin alpha and mu.

ACP6 (acid phosphatase 6, lysophosphatidic) is a protein expressed when myoblast is differentiated into myotubes, and there are no reports related to melasma.

PDZK1 (PDZ domain containing 1) is known to be involved in the binding between membrane proteins such as receptors present in cell membranes and their regulatory proteins, but no association with melasma has been reported to date.

In addition, ACSBG1 (acyl-CoA synthetase bubblegum family member 1), ELOVL3 (elongation of very long chain fatty acids), ELOVL5 (ELOVL family member 5, elongation of long chain fatty acids), C6orf188 (chromosome 6 open reading frame 188) , HSD3B1 (hydroxy-delta-5-steroid dehydrogenase, 3 beta- and steroid delta-isomerase 1), MLSTD1 (male sterility domain containing 1), DKFZP586H2123 (regeneration associated muscle protease), TMEM56 (transmembrane protein 56), ACSM3 (acyl -CoA synthetase medium-chain family member 3) and SLCO4C1 (solute carrier organic anion transporter family, member 4C1) are not known for their exact function, and no association with melasma has been reported. In FLJ21963, only the gene sequence is known, and no other relevant information has been reported.

As a low-expression gene, PCOLCE2 (PROCOLLAGEN C-ENDOPEPTIDASE ENHANCER 2) is a protein involved in cleavage of the propeptide of procollagen 1,2.

H19 is an untranslated mRNA that is maternal imprinted, and its exact function is not known, and no association with melasma has been reported.

GFPT2 (glutamine-fructose-6-phosphate transaminase 2) is a protein that uptakes glucose in the hexoamine pathway, and there are no reports related to melasma.

CCL21 (chemokine (C-C motif) ligand 21) is a cytokine involved in hematopoiesis inhibition.

WIF1 (WNT inhibitory factor 1) is a protein that binds to and inhibits the activity of WNT, a signaling protein that plays an important role in cell-cell interaction in the embryonic stage, and there are no reports related to melasma.

The polynucleotide used as a probe in the diagnostic kit of the present invention includes the full length of a melasma marker gene that is highly expressed or under-expressed in melasma tissue or a fragment thereof. It is preferable that the length of the fragment includes 10 or more contiguous nucleotides, because if the length of the probe is 10 bps or less, it binds non-specifically.

It is preferable that the length of the polynucleotide used as the primer in the diagnostic kit of the present invention is 18-22. This is because if the length of a primer is less than 18bps, it binds non-specifically, and if it exceeds 22bps, the primers bind themselves to each other, resulting in inferior efficiency, and it is unproductive in terms of cost and technology even at the time of manufacture.

In the diagnostic kit of the present invention, a monoclonal antibody having an amino acid sequence encoded by a melasma marker gene as a variable region is prepared by a general method for producing a monoclonal antibody.

Hereinafter, the present invention will be described more specifically with reference to specific examples. The following specific examples are intended to illustrate the present invention and are not intended to be limiting.

<Example 1: Confirmation of stain marker gene>

1. Acquisition and production of tissue samples

The patient's tissue samples were collected from 8 female patients with melasma symptoms. The lesion and normal tissues of the same patient were each biopsied, and the tissues were recovered in sets.

2. From the organization RNA Harvest

Skin tissues of the hyperpigmentation area and normal area were collected from each person about 3mm, put in trizol to purify the RNA, cut into pieces, and then chloroform was added to separate the RNA of the supernatant. To concentrate RNA in the obtained supernatant, an equal amount of isopropanol was added and centrifuged to obtain precipitated RNA, and the precipitated RNA was washed with 70% ethanol for base removal. After dissolving the obtained RNA in purified water, electrophoresis was performed to measure the purity and quantity to determine whether the bands of 28S and 18S rRNA are clearly distinguished, and then the purity is measured, and the amount is measured by the thickness of the band. I did.

3. For microarray Preparation of the sample

Microarray was performed to compare the expression patterns of mRNA obtained from each sample, and Biosystems Chemiluminescent RT-IVT Labeling Kit (ABI) was used to synthesize cRNA. The method is as follows. After adding the same amount of RNA obtained from each sample, cDNA is synthesized using reverse transcriptase, and then cRNA is synthesized from the synthesized cDNA (in vitro transcription). At this time, the synthesized cRNA is labeled by inserting the UTP labeled with DIG. After measuring the purity and amount of the labeled cRNA, it was combined with Applied Biosystems 1700 human chip (ABI) in which a total of 1700 genes were spotted. In order to confirm the bound cRNA, an antibody that specifically binds to DIG was added and reacted. At this time, since a chemiluminescent enzyme is bound to the antibody, a chemiluminescent substrate is added and reacted to obtain a photosensitive image. The intensity of the obtained image was analyzed and statistically processed to identify genes that were specifically high and low expressed in the melasma region. Among the total 1700 genes, 23 kinds of genes were highly expressed (Table 1), and 5 kinds were low-expressed genes (Table 2).

4. RT - PCR

Each 1 μg of RNA collected from the epidermis of the normal and lesion site of the melasma patient was reacted with oligo-dT primer (24 mer) 100 pmol (Bioneer) at 70° C. for 10 minutes (total volume: 20 μl). Then, 4 µl of 5x first chain reaction buffer, 2 µl of 0.1M DTT, 4 µl of 2.5 mM dNTP mix, and 1 µl of SuperscriptII (all invitrogen) were added to react at 42° C. for 1 hour, and finally reacted at 70° C. for 10 minutes. And elongate to prepare a cDNA template. 5 µl of 10x buffer for rTaq, _{3 µl of 25 mM MgCl 2} , 5 µl of 2.0 mM dNTP mix, 0.5 µl of rTaq (all Bioneer), 33.5 µl of ddH2 0, 1 µl each of cDNA template, primer sense and antisense (see Table 3 below) ) PCR reaction [94℃ for 2 minutes, (94℃ for 30 seconds, 55℃ for 30 seconds, 72℃ for 1 minute) for 30 cycles, 72℃ for 10 minutes] by adding 1 µl of each 20 mM (total volume 50 µl) I did.

Genetic symbol Primer type Primer sequence H19
Sense (SEQ ID NO: 1) aaa gac acc atc gga aca gc Antisense (SEQ ID NO: 2) aga gtc gtg gag gct ttg aa PCOLCE2
Sense (SEQ ID NO: 3) gag gca aaa tca tgc caa ac Antisense (SEQ ID NO: 4) gag gcc tca tac ctc cat ca WIF1
Sense (SEQ ID NO: 5) att ggg caa aaa tgc gta ag Antisense (SEQ ID NO: 6) ttg ttt aaa tgc cac tac cac a SOAT1
Sense (SEQ ID NO: 7) cga ata tgc ctt ggc tgt tt Antisense (SEQ ID NO: 8) gac tcc att gcc caa gaa aa PDZK1
Sense (SEQ ID NO: 9) gag gct cca gct cct act cc Antisense (SEQ ID NO: 10) cgc cct tct gta cct ctt tg THRSP
Sense (SEQ ID NO: 11) aga gaa tgg aac cgc aga ga Antisense (SEQ ID NO: 12) act tgt ccc gtc att tcc tg TMEM56
Sense (SEQ ID NO: 13) agc aaa agt ttc ccc agg tt Antisense (SEQ ID NO: 14) cac gtt tgc aag tga tgg ac CYR61
Sense (SEQ ID NO: 15) cag ctg acc agg act gtg aa Antisense (SEQ ID NO: 16) gtc tag gtg tgc cct gga aa

Finally, a band of about 100 bp was obtained by electrophoresis with an agarose gel containing 1.5% SYBR GREEN, and the results are shown in FIG. 1. GAPDH was shown as a control. 1, the H19, PCOLCE2, WIF1, SOAT1 genes are significantly lowered in expression in the epidermal melasma region than in the normal region, and PDZK1, THRSP, TMEM56, CYR61 are relatively increased in expression in the melasma region. Can be seen, which is consistent with the microarray results.

5. Real Time PCR

In order to reconfirm the above results, 2 µg of each RNA collected from the epidermis of the normal and lesion site of 12 melasma patients was reacted with 100 pmol (applied biosystems) of oligo-dT primer for 5 minutes (total dose 5 µl). . Then, 4 µl of 5x first chain reaction buffer _{, 2.4 µl of MgCl 2} , 1 µl of 2.5 mM dNTP mix, and 1 µl of Rtase were added to react at 42°C for 1 hour, and finally reacted at 70°C for 15 minutes to elongate cDNA. A template was prepared. 25 μl of TaqMan Universal PCR Master 2x buffer containing AmpliTaq Gold® DNA polymerase, 22.5 μl each of cDNA template, primer sense and antisense (see Table 4 below) mix 20 mM 2.5 μl (total volume 50 μl) for PCR Reaction [95°C for 10 minutes, (95°C for 15 seconds, 60°C for 1 minute) 40 cycles, 72°C for 10 minutes] was carried out.

Genetic symbol Primer type Primer sequence H19
Sense (SEQ ID NO: 17) atg aca tgg tcc ggt gtg a Antisense (SEQ ID NO:18) aga aac aga ccc gct tct tg PDZK1
Sense (SEQ ID NO: 19) ggc agg ctc aga aca gaa ag Antisense (SEQ ID NO: 20) cta cca gat cat tgt tct tca

The results are shown in FIG. 2. According to FIG. 2, it can be seen that the expression level of each mRNA in the melasma region of the melasma patient is consistent with the microarray result by decreasing H19 and increasing PDZK1 compared to the normal region.

As described above, according to the present invention, it is possible to sensitively and quickly predict the possibility of spot formation in the skin by using 23 kinds of genes and 5 kinds of low-expression genes that are specifically highly expressed at the melasma lesion site as melasma marker genes. .

Attach electronic file of sequence list

Claims (14)

A polynucleotide or a fragment thereof said polynucleotide is a polynucleotide encoding WIF1, said fragment comprising at least one polynucleotide comprising at least 10 contiguous nucleotides,
Or a complementary polynucleotide thereof; And
And a solid support on which the polynucleotide is bound to the surface,
If the amount of the transcript hybridized to the polynucleotide is less than the normal value, the diagnostic kit for predicting blemish formation of the skin, characterized in that it is diagnosed that the skin is likely to form blemishes. The method of claim 1,
The kit includes F7 [coagulation factor VII], MGC34923 (virtual protein MGC34923), FLJ21963 (FLJ21963 protein), PRKAA1 [protein kinase AMP-activated alpha 1 catalytic subunit. subunit)], ELOVL5 (ELOVL family member 5), ACSM3 [acyl-CoA synthetase medium-chain family member 3]], DGAT2L3 [diacylglycerol O-acyltransferase 2-like 3 (diacylglycerol O-acyltransferase 2-like 3)], GABRA4 [gamma-aminobutyric acid A receptor alpha 4], TMEM56 [transmembrane protein 56], SLCO4C1 [solute [Solute carrier organic anion transporter family member 4C1], THRSP [THYROID HORMONE-RESPONSIVE SPOT14], HAO2 [hydroxyacid oxidase 2] , IGJ [Immunoglobulin J Paul Peptide (immunoglobulin J polypeptide)], ACSBG1 [acyl-CoA synthetase bubblegum family member 1], ACP6 [acid phosphatase 6], SOAT1 [acylco A: cholesterol acyltransferase (AC), ELOVL3 [elongation of very long chain faatty acids], C6orf188 [chromosome 6 open reading frame 188] reading frame 188)], HSD3B1 [hydroxy-delta-5-steroid dehydrogenase 3 beta and steroid delta-isomerase 1], MLSTD1 [male sterility domain containing 1], Cyr61 [CYSTEINE-RICH PROTEIN 61], and DKFZP586H2123 [regeneration associated muscle protease] Selection As a further polynucleotide, or fragment thereof encoding at least one, the segment is at least one comprising at least 10 consecutive nucleotides polynucleotide,
Or a complementary polynucleotide thereof; And
It further comprises a solid support that binds the polynucleotide to the surface,
If the amount of the transcript hybridized to the additional polynucleotide is greater than the normal value, the diagnostic kit for predicting blemish formation of the skin, characterized in that it is diagnosed that the skin is likely to form blemishes. The method of claim 1,
The kit includes PCOLCE2 [procollagen C-endopeptidase enhancer 2], GFPT2 [glutamine-fructose-6-phosphate transaminase 2] , And an additional polynucleotide or fragment thereof encoding at least one selected from the group consisting of CCL21 (chemokine CC motif ligand 21), wherein the fragment comprises at least 10 contiguous nucleotides; ,
Or a complementary polynucleotide thereof; And
It further comprises a solid support that binds the polynucleotide to the surface,
If the amount of the transcript hybridized to the additional polynucleotide is less than the normal value, the diagnostic kit for predicting blemish formation of the skin, characterized in that it is diagnosed that the skin is likely to form blemishes. At least one polynucleotide comprising 18-22 consecutive nucleotides as a fragment of a polynucleotide encoding WIF1; And
A fragment of a polynucleotide complementary to said polynucleotide, comprising one or more polynucleotides comprising 18-22 contiguous nucleotides,
If the amount of DNA amplified by using the polynucleotides as a primer is less than the normal value for diagnosing the skin dysplasia diagnostic kit, characterized in that it is diagnosed that the skin is likely to form. 5. The method of claim 4,
The kit is composed of F7, MGC34923, FLJ21963, PRKAA1, ELOVL5, ACSM3, DGAT2L3, GABRA4, TMEM56, SLCO4C1, THRSP, HAO2, IGJ, ACSBG1, ACP6, SOAT1, ELOVL3, C6orf188, HSDDH1, C2F, ZL, D, D, D, D, DK, DK, F, H, D, D, D, D, D, D, DK, D, H, D, D, D, D, B, D, D, B, D, D, D, B, D, D, D, B, D, D, and B, Dl, F, D, H, D, D, D, D, D, D, D, D, D, C, D, D, B, D, and B At least one polynucleotide comprising 18-22 contiguous nucleotides as fragments of additional polynucleotides encoding those selected from; And
A fragment of a polynucleotide complementary to said polynucleotide, further comprising one or more polynucleotides comprising 18-22 contiguous nucleotides,
If the amount of DNA amplified by using the additional polynucleotides amplified more than the normal value, the diagnostic kit for predicting blemish formation of the skin, characterized in that it is diagnosed that the skin is likely to form blemishes. 5. The method of claim 4,
The kit comprises one or more polynucleotides comprising 18-22 contiguous nucleotides as fragments of additional polynucleotides encoding those selected from the group consisting of PCOLCE2, GFPT2, and CCL21; And
A fragment of a polynucleotide complementary to a polynucleotide selected from the group, further comprising one or more polynucleotides comprising 18-22 contiguous nucleotides,
If the amount of DNA amplified by using the additional polynucleotides less than the normal value of the diagnostic kit for predicting the appearance of spots of the skin, characterized in that it is diagnosed that the skin is likely to form a spot. Includes monoclonal antibodies that bind to WIF1 antigen,
If the amount of the antigen bound to the antibody is less than the normal value, the diagnostic kit for predicting blemish formation of the skin, characterized in that it is diagnosed that the skin is likely to form blemishes. The method of claim 7, wherein
The kit is composed of F7, MGC34923, FLJ21963, PRKAA1, ELOVL5, ACSM3, DGAT2L3, GABRA4, TMEM56, SLCO4C1, THRSP, HAO2, IGJ, ACSBG1, ACP6, SOAT1, ELOVL3, C6orf188, HSDDH1, C2F, ZL, D, D, D, D, DK, DK, F, H, D, D, D, D, D, D, DK, D, H, D, D, D, D, B, D, D, B, D, D, D, B, D, D, D, B, D, D, and B, Dl, F, D, H, D, D, D, D, D, D, D, D, D, C, D, D, B, D, and B An additional monoclonal antibody that binds to an antigen selected from
If the amount of the antigen bound to the additional antibody is greater than the normal value for diagnosing the formation of blemishes for the skin, characterized in that the diagnosis is likely to have a blemish. The method of claim 7, wherein
The kit includes additional monoclonal antibodies that bind to an antigen selected from the group consisting of PCOLCE2, GFPT2, and CCL21,
If the amount of the antigen bound to the additional antibody is less than the normal value, the diagnostic kit for predicting blemish formation of the skin, characterized in that it is diagnosed that the skin is likely to form blemishes. A method for predicting the formation of spots on the skin, wherein the method for predicting the appearance of spots on the skin is determined to be a skin having a chance of spotting if the expression level of the WIF1 gene in the epidermis is less than the normal value. The method of claim 10,
The method is F7, MGC34923, FLJ21963, PRKAA1, ELOVL5, ACSM3, DGAT2L3, GABRA4, TMEM56, SLCO4C1, THRSP, HAO2, IGJ, ACSBG1, ACP6, SOAT1, ELOVL3, C6orf188F, ZDH3, HSD3, ZD3, HSD3, HSD3, HSD3 Further comprising measuring the expression level of one or more additional genes selected from the group consisting of,
If the expression level of the additional gene is greater than the normal value, it is judged that the skin is likely to form a blemishes. The method of claim 10,
The method further comprises measuring the expression level of one or more additional genes selected from the group consisting of PCOLCE2, GFPT2, and CCL21 in the epidermis,
If the expression level of the additional gene is less than the normal value, it is judged that the skin is likely to form a blemishes, characterized in that the method for preventing skin formation. As a method of screening anti-glare inhibitors,
Binding the test compound to a protein encoded by the WIF1 gene; And
Determining whether the test compound promotes or inhibits the action of the protein. 14. The method of claim 13,
The method the group consisting of F7, MGC34923, FLJ21963, PRKAA1, ELOVL5, ACSM3, DGAT2L3, GABRA4, TMEM56, SLCO4C1, THRSP, HAO2, IGJ, ACSBG1, ACP6, SOAT1, ELOVL3, C6orf188, HSD3B1, MLSTD1, Cyr61, and DKFZP586H2123 Binding the test compound to an additional protein encoded by one or more genes selected from, or an additional protein encoded by one or more genes selected from the group consisting of PCOLCE2, GFPT2, and CCL21; And
The method of claim 1, further comprising identifying whether the test compound promotes or inhibits the action of the additional protein.

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