BIOMARKERS FOR DIAGNOSING KELOID SKIN OR KELOID SCAR, AND USE THEREOF

Abstract

The present invention relates to: biomarker proteins for diagnosing scars; a composition for diagnosing scars by using the same; a biochip for diagnosing scars; and a method for obtaining information on the scars. Proteins identified by the present invention can be applied as biomarkers for screening scars. A patient with keloid scars can be diagnosed by using the biomarker. In addition, the biomarker can be used for treating a patient with keloid scars.

Claims

1. A composition for diagnosing keloid skin or a keloid scar, comprising: any one or more peptides selected from the group consisting of the peptides disclosed in the GenPept database shown in the following table and thereof. TABLE-US-00005 GenPept No Protein name Accession no. 1 26S proteasome non-ATPase regulatory subunit 13 NP_002808 isoform 1 2 40S ribosomal protein S12 NP_001007 3 Apo Form Of Human S100a16 3NXA_A 4 ASPRV1 protein AAH31997 5 bifunctional purine biosynthesis protein PURH NP_004035.2 6 Cellular Retinoic Acid Binding Protein II In Complex 2CBS_A With A Synthetic Retinotic Acid 7 Aldehyde Reductase 2ALR_A 8 Human Epidermal Fatty Acid-binding Protein (fabp5) In 4AZM_A Complex With The Inhibitor Bms-309413 9 Prolyl Oligopeptidase With Gsk552 3DDU_A 10 Solution Structure Of Apo S100a16 2L50_A 11 Structure Of [r563a] Leukotriene A4 Hydrolase ISQM_A 12 The High Resolution Structure Of Annexin III Shows 1AXN_A Differences With Annexin V 13 Three Crystal Structures Of Human Coactosin-Like 1T2L_A Protein 14 cystatin-B NP_000091 15 Ezrin AAH68458 16 Isopentenyl-diphosphate Delta-Isomerase 1; Short = IPPI1 Q13907 17 keratin, type I cytoskeletal 17 NP_000413 18 macrophage-capping protein isoform 9 XP_515584 19 mitochondrial ATP synthase, H+ transporting F1 ABD77240 complex beta subunit 20 NCOR1 protein, partial AAH58511 21 Phosphoglycerate mutase 1 (brain) AAH62302 22 PREDICTED: tublin alpha-1B chain-like isoform 2 XP_002823231 23 protein S100-A8 isoform 4 [Pan troglodytes] XP_001138065 24 R33729_1 AAC27824 25 TALDO1 protein AAH18847 26 type 1 keratin 16 AAB35421 27 all taxonomy (Crystal Structure Of Bovine Serum 3V03_A Albumin) 28 alpha-actinin-4 NP_004915 29 alpha-crystallin B chain NP_001876 30 Crystal Structure Of Human Enolase 1 3B97_A 31 Human Peroxiredoxin 5 1HD2_A 32 Structural And Electrophysiological Analysis Of Annexin 1HVE_A V Mutants, Mutagenesis Of Human Annexin V 33 Structure Of S100a4 In Complex With Non-Muscle 4ETO_A Myosin-IIa Peptide 34 X-Ray Crystal Structure Of Human Galectin-1 1GZW_B 35 Structure Of The Human Class 1 Histocompatibility 1HLA_M Antigen, Hla-A2 36 Chip-Ubc13-Uevla Complex 2C2V_B 37 chorionic somatomammotropin hormone-like 1 isoform 3 NP_001309 38 dynactin 3 (p22) CAI13144 39 Eukaryotic translation initiation factor 2B, subunit 2 AAH00494 beta, 39 kDa 40 gelsolin isoform b NP_937895 41 glutaredoxin-3 NP_006532 42 glutathione S-tranferase P NP_000843 43 heat shock cognate 71 kDa protein isoform 1 NP_006588 44 HMOX2 CAG33041 45 Human Glutathione Transferase Omega 1, Delta 155 3LFL_A 46 keratin 1, keratin, type I cytoskeletal 9 AAG41947 47 LMNB1 protein AAH78178 48 Moesin Ferm Domain Bound To Ebp50 C-Terminal 1SGH_A Peptide 49 Mrp14 Complexed With Chaps 1IRJ_A 50 MYO5C protein AAH64841 51 nicotinamide N-methyltransferase NP_006160 52 Nucear transfactor 2 NP_005787 53 nucleoside diphosphate kinase A isoform a NP_937818 54 O-Methyltransferase 3BWM_A 55 platelet-activating factor acetylhydrolase IB subunit NP_002564 gamma 56 protein S100-A11 NP_005611 57 pyruvate dehydrogenase (lipoamide) beta EAW65372 58 serine/threonine-protein phosphatase 2A catalytic NP_058736, subunit beta isoform 59 tubulin alpha-1B chain isoform 2 XP_002823231 60 tumor necrosis factor type 1 receptor associated A55877 protein TRAP-1 - human 61 ubiquitin carboxyl-terminal hydrolase 14 isoform a NP_005142 62 vacuolar protein sorting-associated protein 29 NP_057310 63 X-ray repair cross-complementing protein 5 NP_066964 64 Chain A, Structure Of S100a4 In Complex With 4ETO_A Non-Muscle Myosin-IIa Peptide 65 Phosphoglycerate mutase 1 (brain) AAH62302 66 putative G-protein coupled receptor BAB89334 67 Chain A, Human Quinone Reductase Type 2 1QR2_A 68 T-plastin polypeptide AAB02844 69 FLNA protein AAH14654 70 transgelin variant BAD92792 71 hCG38213, isoform CRA_d EAW76181 72 leukotriene A4 hydrolase, isoform CRA_a EAW97557 73 prolyl 4-hydroxylase subunit alpha-1 Isoform 2 NP_001017962 precursor 74 ubiquitin carboxyl-terminal hydrolase isozyme L1 NP_004172 75 rho GDP-dissociation inhibitor 1 isoform a NP_004300 76 Cytokeratin-14 P02533 77 Keratin 5 AAH24292 78 prohibitin AAS88903 79 FLJ00410 protein BAC03467 80 serpin B5 NP_002630

2. A biochip for diagnosing keloid skin or a keloid scar, comprising: the composition of claim 1.

3. A method for obtaining information on keloid skin or a keloid scar from a subject, comprising: (a) measuring one or more biomarkers in a biological sample obtained from a subject, wherein the one or more biomarkers are selected from the composition of claim 1; and (b) correlating the measured value(s) with keloid skin or a keloid scar as compared with a normal person.

4. A method for obtaining information on keloid skin or a keloid scar from a subject, comprising: (a) measuring one or more biomarkers in a biological sample obtained from a subject, wherein the one or more biomarkers are selected from the composition of claim 1; and (b) correlating the measured value(s) with keloid skin or a keloid scar as compared with a normal person, wherein, in Step (b), when measured values of any one or more peptides selected from the group consisting of the peptides 1 to 26 and 64 to 75 shown in the table of claim 1 are higher than those of a normal person, the values are correlated with keloid skin or a keloid scar.

5. A method for obtaining information on keloid skin or a keloid scar from a subject, comprising: (a) measuring one or more biomarkers in a biological sample obtained from a subject, wherein the one or more biomarkers are selected from the composition of claim 1, and (b) correlating the measured value(s) with keloid skin or a keloid scar as compared with a normal person, wherein, in Step (b), when measured values of any one or more peptides selected from the group consisting of the peptides 27 to 63 and 76 to 80 shown in the table of claim 1 are lower than those of a normal person, the values are correlated with keloid skin or a keloid scar.

6. A pharmaceutical composition for preventing or treating keloid skin or a keloid scar, comprising: a peptide inhibitor that inhibits the function of any one selected from the group consisting of the peptides 1 to 26 and 64 to 75 shown in the table below or the expression of a gene encoding the peptide, as an active ingredient, or a material that increases the expression of any one selected from the group consisting of the peptides 27 to 63 and 76 to 80 shown in the table below, as an active ingredient. TABLE-US-00006 GenPept No Protein name Accession no. 1 26S proteasome non-ATPase regulatory subunit 13 NP_002808 isoform 1 2 40S ribosomal protein S12 NP_001007 3 Apo Form Of Human S100a16 3NXA_A 4 ASPRV1 protein AAH31997 5 bifunctional purine biosynthesis protein PURH NP_004035.2 6 Cellular Retinoic Acid Binding Protein II In Complex 2CBS_A With A Synthetic Retinoic Acid 7 Aldehyde Reductase 2ALR_A 8 Human Epidermal Fatty Acid-binding Protein (fabp5) In 4AZM_A Complex With The Inhibitor Bms-309413 9 Prolyl Oligopeptidase With Gsk552 3DDU_A 10 Solution Structure Of Apo S100a16 2L50_A 11 Structure Of [r563a] Leukotriene A4 Hydrolase 1SQM_A 12 The High Resolution Structure Of Annexin III Shows 1AXN_A Differences With Annexin V 13 Three Crystal Structures Of Human Coactosin-Like 1T2L_A Protein 14 cystatin-B NP_000091 15 Ezrin AAH68458 16 Isopentenyl-diphosphate Delta-isomerase 1; Short = IPPI1 Q13907 17 keratin, type I cytoskeletal 17 NP_000413 18 macrophage-capping protein isoform 9 XP_515584 19 mitochondrial ATP synthase, H+ transporting F1 ABD77240 complex beta subunit 20 NCOR1 protein, partial AAH58511 21 Phosphoglycerate mutase 1 (brain) AAH62302 22 PREDICTED: tublin alpha-1B chain-like isoform 2 XP_002823231 23 protein S100-A8 isoform 4 [Pan troglodytes] XP_001138065 24 R33729_1 AAC27824 25 TALDO1 protein AAH18847 26 type I keratin 16 AAB35421 27 all taxonomy (Crystal Structure Of Bovine Serum 3V03_A Albumin) 28 alpha-actinin-4 NP_004915 29 alpha-crystallin B chain NP_001876 30 Crystal Structure Of Human Enolase 1 3B97_A 31 Human Peroxiredoxin 5 1HD2_A 32 Structural And Electrophysiological Analysis Of Annexin 1HVE_A V Mutants, Mutagenesis Of Human Annexin V 33 Structure Of S100a4 In Complex With Non-Muscle 4ETO_A Myosin-IIa Peptide 34 X-Ray Crystal Structure Of Human Galectin-1 1GZW_B 35 Structure Of The Human Class 1 Histocompatibility 1HLA_M Antigen, Hla-A2 36 Chip-Ubc13-Uevla Complex 2C2V_B 37 chorionic somatomammotropin hormone-like 1 isoform 3 NP_001309 38 dynactin 3 (p22) CAI13144 39 Eukaryotic translation initiation factor 2B, subunit 2 AAH00494 beta, 39 kDa 40 gelsolin isoform b NP_937895 41 glutaredoxin-3 NP_006532 42 glutathione S-tranferase P NP_000843 43 heat shock cognate 71 kDa protein isoform 1 NP_006588 44 HMOX2 CAG33041 45 Human Glutathione Transferase Omega 1, Delta 155 3LFL_A 46 keratin 1, keratin, type I cytoskeletal 9 AAG41947 47 LMNB1 protein AAH78178 48 Moesin Ferm Domain Bound To Ebp50 C-Terminal ISGH_A Peptide 49 Mrp14 Complexed With Chaps 1IRJ_A 50 MYO5C protein AAH64841 51 nicotinamide N-methyltransferase NP_006160 52 Nucear transfactor 2 NP_005787 53 nucleoside diphosphate kinase A isoform a NP_937818 54 O-Methyltransferase 3BWM_A 55 platelet-activating factor acetylhydrolase IB subunit NP_002564 gamma 56 protein S100-A11 NP_005611 57 pyruvate dehydrogenase (lipoamide) beta EAW65372 58 serine/threonine-protein phosphatase 2A catalytic NP_058736, subunit beta isoform 59 tubulin alpha-1B chain isoform 2 XP_002823231 60 tumor necrosis factor type 1 receptor associated A55877 protein TRAP-1 - human 61 ubiquitin carboxyl-terminal hydrolase 14 isoform a NP_005142 62 vacuolar protein sorting-associated protein 29 NP_057310 63 X-ray repair cross-complementing protein 5 NP_066964 64 Chain A, Structure Of S100a4 In Complex With 4ETO_A Non-Muscle Myosin-IIa Peptide 65 Phosphoglycerate mutase 1 (brain) AAH62302 66 putative G-protein coupled receptor BAB89334 67 Chain A, Human Quinone Reductase Type 2 1QR2_A 68 T-plastin polypeptide AAB02844 69 FLNA protein AAH14654 70 transgelin variant BAD92792 71 hCG38213, isoform CRA_d EAW76181 72 leukotriene A4 hydrolase, isoform CRA_a EAW97557 73 prolyl 4-hydroxylase subunit alpha-1 isoform 2 NP_001017962 precursor 74 ubiquitin carboxyl-terminal hydrolase isozyme L1 NP_004172 75 rho GDP-dissociation inhibitor 1 isoform a NP_004300 76 Cytokeratin-14 P02533 77 Keratin 5 AAH24292 78 prohibitin AAS88903 79 FLJ00410 protein BAC03467 80 serpin B5 NP_002630

7. The pharmaceutical composition of claim 6, wherein the inhibitor is an aptamer, a small compound, an antibody or a functional fragment of the antibody, which regulates the expression of any one selected from the peptides 1 to 26 and 64 to 75, or a viral vector, a non viral vector, an antisense oligonucleotide, miRNA, siRNA or shRNA that regulates the expression of a gene encoding the peptide.

8. The pharmaceutical composition of claim 6, wherein the material that increases the expression of any one selected from the group consisting of the peptides 27 to 63 and 76 to 80 shown in the tables is a peptide selected from the group consisting of the peptides 27 to 63 and 76 to 80, a gene encoding the peptide, or a viral vector, non-viral vector, protein or small-molecule compound regulating the expression of the peptide.

9. A quasi-drug composition for preventing or improving keloid skin or a keloid scar, comprising: a peptide inhibitor that inhibits the function of any one selected from the group consisting of the peptides 1 to 26 and 64 to 75 shown in the table below or the expression of a gene encoding the peptide, as an active ingredient, or a material that increases the expression of any one selected from the group consisting of the peptides 27 to 63 and 76 to 80 shown in the table below, as an active ingredient. TABLE-US-00007 GenPept No Protein name Accession no. 1 26S proteasome non-ATPase regulatory subunit 13 NP_002808 Isoform 1 2 40S ribosomal protein S12 NP_001007 3 Apo Form Of Human S100a16 3NXA_A 4 ASPRV1 protein AAH31997 5 bifunctional purine biosynthesis protein PURH NP_004035.2 6 Cellular Retinoic Acid Binding Protein II In Complex 2CBS_A With A Synthetic Retinoic Acid 7 Aldehyde Reductase 2ALR_A 8 Human Epidermal Fatty Acid-binding Protein (fabp5) In 4AZM_A Complex With The Inhibitor Bms-309413 9 Prolyl Oligopeptidase With Gsk552 3DDU_A 10 Solution Structure Of Apo S100a16 2L50_A 11 Structure Of [r563a] Leukotriene A4 Hydrolase 1SQM_A 12 The High Resolution Structure Of Annexin III Shows 1AXN_A Differences With Annexin V 13 Three Crystal Structures Of Human Coactosin-Like 1T2L_A Protein 14 cystatin-B NP_000091 15 Ezrin AAH68458 16 Isopentenyl-diphosphate Delta-Isomerase 1; Short = IPPI1 Q13907 17 keratin, type I cytoskeletal 17 NP_000413 18 macrophage-capping protein isoform 9 XP_515584 19 mitochondrial ATP synthase, H+ transporting F1 ABD77240 complex beta subunit 20 NCOR1 protein, partial AAH58511 21 Phosphoglycerate mutase 1 (brain) AAH62302 22 PREDICTED: tubulin alpha-1B chain-like isoform 2 XP_002823231 23 protein S100-A8 isoform 4 [Pan troglodytes] XP_001138065 24 R33729_1 AAC27824 25 TALDO1 protein AAH18847 26 type I keratin 16 AAB35421 27 all taxonomy (Crystal Structure Of Bovine Serum 3V03_A Albumin) 28 alpha-actinin-4 NP_004915 29 alpha-crystallin B chain NP_001876 30 Crystal Structure Of Human Enolase 1 3B97_A 31 Human Peroxiredoxin 5 1HD2_A 32 Structural And Electrophysiological Analysis Of Annexin 1HVE_A V Mutants, Mutagenesis Of Human Annexin V 33 Structural Of S100a4 In Complex With Non-Muscle 4ETO_A Myosin-IIa Peptide 34 X-Ray Crystal Structure Of Human Galectin-1 1GZW_B 35 Structure Of The Human Class I Histocompatibility 1HLA_M Antigen, Hla-A2 36 Chip-Ubc13-Uev1a Complex 2C2V_B 37 chorionic somatomammotropin hormone-like 1 isoform 3 NP_001309 38 dynactin 3 (p22) CAI13144 39 Eukaryotic translation initiation factor 2B, subunit 2 AAH00494 beta, 39 kDa 40 gelsolin isoform b NP_937895 41 glutaredoxin-3 NP_006532 42 glutathione S-transferase P NP_000843 43 heat shock cognate 71 KDa protein isoform 1 NP_006588 44 HMOX2 CAG33041 45 Human Glutathione Transferase Omega 1, Delta 155 3LFL_A 46 keratin 1, keratin, type I cytoskeletal 9 AAG41947 47 LMNB1 protein AAH78178 48 Moesin Ferm Domain Bound To Ebp50 C-Terminal 1SGH_A Peptide 49 Mrp14 Complexed With Chaps 1IRJ_A 50 MYO5C protein AAH64841 51 nicotinamide N-methyltransferase NP_006160 52 Nucear transfactor 2 NP_005787 53 nucleoside diphosphate kinase A isoform a NP_937818 54 O-Methyltransferase 3BWM_A 55 platelet-activating factor acetylhydrolase IB subunit NP_002564 gamma 56 protein S100-A11 NP_005611 57 pyruvate dehydrogenase (lipoamide) beta EAW65372 58 serine/threonine-protein phosphatase 2A catalytic NP_058736, subunit beta isoform 59 tubulin alpha-1B chain isoform 2 XP_002823231 60 tumor necrosis factor type 1 receptor associated A55877 protein TRAP-1 - human 61 ubiquitin carboxyl-terminal hydrolase 14 isoform a NP_005142 62 vacuolar protein sorting-assodated protein 29 NP_057310 63 X-ray repair cross-complementing protein 5 NP_066964 64 Chain A, Structure Of S100a4 In Comptex With 4ETO_A Non-Muscle Myosin-IIa Peptide 65 Phosphoglycerate mutase 1 (brain) AAH62302 66 putative G-protein coupled receptor BAB89334 67 Chain A, Human Quinone Reductase Type 2 1QR2_A 68 T-plastin polypeptide AAB02844 68 FLNA protein AAH14654 70 transgelin variant BAD92792 71 hCG38213, isoform CRA_d EAW76181 72 leukotriene A4 hydrolase, isoform CRA_a EAW97557 73 prolyl 4-hydroxylase subunit alpha-1 isoform 2 NP_001017962 precursor 74 ubiquitin carboxyl-terminal hydrolase isozyme L1 NP_004172 75 rho GDP-dissociation inhibitor 1 isoform a NP_004300 76 Cytokeratin-14 P02533 77 Keratin 5 AAH24292 78 prohibitin AAS88903 79 FLJ00410 protein BAC03467 80 serpin B5 NP_002630

Description

DESCRIPTION OF DRAWINGS

[0085] FIG. 1 is a 2D electrophoresis image for proteins extracted from keratinocytes isolated from normal skin tissue.

[0086] FIG. 2 is a 2D electrophoresis image for proteins extracted from keratinocytes isolated from skin tissue in a keloid scar region. FIG. 3 is a 2D electrophoresis image for proteins extracted from fibroblasts isolated from normal skin tissue.

[0087] FIG. 4 is a 2D electrophoresis image for proteins extracted from fibroblasts isolated from skin tissue in a keloid scar region.

MODES OF THE INVENTION

[0088] Hereinafter, the present invention will be described in further detail with reference to examples. These examples are merely provided to illustrate the present invention, and it will be apparent to those of ordinary skill in the art that it should not be construed that the scope of the present invention is limited by the following examples.

EXAMPLE 1

Culture of Keratinocytes and Fibroblasts

[0089] 1-1: Culture of Keratinocytes

[0090] Keratinocytes isolated from normal skin tissue and skin tissue in scar regions of patients with keloid scars were cultured on a feeder in a DMEM/F12 medium containing 10% fetal bovine serum (FBS) and 10 ng/ml EGF. When the cultured cells were grown to 70 to 80%, a feeder was removed and only the keratinocytes were isolated.

[0091] 1-2: Culture of Fibroblasts

[0092] Fibroblasts isolated from normal skin tissue and skin tissue in scar regions of patients with keloid scars were cultured in an F12 medium containing 10% FBS. When the cultured cells were grown to 70 to 80%, only the fibroblasts were isolated.

EXAMPLE 2

Protein Extraction

[0093] A 10 volume of a sample solution consisting of 7M urea, 2M thiourea, 4%(w/v) 3-[(3-cholamidopropyl)dimethyammonio]-1-propanesulfonate (CHAPS), 1%(w/v) dithiothreitol (DTT), 2%(v/v) pharmalyte and 1 mM benzamidine was prepared. The keratinocytes or fibrocytes isolated in Example 1 were mixed with the prepared sample solution, and then the cells were disrupted using a homogenizer. In addition, for protein extraction, the cells were vortexed for 1 hour, centrifuged at 15 C. and 15,000 rpm for 1 hour, and then the supernatant was used as a sample for 2D electrophoresis.

EXAMPLE 3

Protein Expression Analysis using 2D Electrophoresis

[0094] For primary isoelectric focusing (IEF), IPG strips were reswelled with a reswelling solution consisting of 7M urea, 2M thiourea, 2% 3-[(3-cholamidopropyl)dimethyammonio]-1-propanesulfonate (CHAPS), 1% DTT and 1% pharmalyte at room temperature for approximately 12 to 16 hours. 200 g of the sample per strip was used, and subjected to IEF at 20 C. For IEF, the voltage was increased from 150V to 3,500V within 3 hours, and the voltage was maintained at 3,500V for 26 hours, and then the focusing was completed after 96 kVh.

[0095] Before SDS-PAGE was performed secondarily, IPG strips were incubated in a 1% DTT-containing equilibration buffer (50 mM Tris-Cl, pH 6.8, 6M urea, 2% SDS, and 30% glycerol) for 10 minutes, and then further incubated in a 2.5% iodoacetamide-containing equilibration buffer for 10 minutes. Equilibrated strips were arranged on SDS-PAGE gels (2024 cm, 10 to 16%), and run at 1.7 kVh and 20 C. using a Hoefer DALT 2D system. After 2D electrophoresis was completed, proteins on the 2D gels were visualized by silver staining, and a glutaraldehyde treatment step was omitted for protein identification by mass spectrometry. The silver-stained 2D gel was scanned, and stored in the form of a file with an extension, TIFF. FIG. 1 shows the 2DE image of normal keratinocytes, FIG. 2 shows the 2DE image of keloidal keratinocytes, FIG. 3 shows the 2DE image of normal fibroblasts, and FIG. 4 shows the 2DE image of keloidal fibroblasts.

[0096] A quantitative analysis for checking changes in protein spot expression from the scanned images was performed using PDQuest software. By comparing spots of the normal cell population and spots of the keloidal cell population, spots 2-fold or more increased or decreased compared to the normal cells were selected, and the result of performing protein identification for the selected spots are shown in the following Tables 3 (the proteins extracted from the keratinocytes) and 4 (the proteins extracted from the fibroblasts). Nos. 1 to 26 and 64 to 75 represent the proteins 2-fold or more increased compared to the normal cells, and Nos. 27 to 63 and 76 to 80 represent the proteins 2-fold or more decreased compared to the normal cells.

[0097] Protein peptide mass fingerprinting (PMF) identification using MALDI-TOF was performed through a known conventional method (Biomol Ther (Seoul), May 30, 2013; 21(3): 190-195). As a mass spectrometer, Microflex LRF 20 (Bruker Daltonics) was used. Protein fragments dropped on a target plate were vaporized by N2 laser irradiation at 337 nm and accelerated by a 20 Kv injection pulse. A mass spectrum for each protein spot was obtained by a cumulative peak of 300 laser shots. For analysis of the mass spectrum, an ion peak m/z (842.510, 2211.1046) of a peptide produced by the autolysis of trypsin was used as a standard peak. For protein identification from the analyzed mass spectrum, a MASCOT search engine (http://www.matrixscience.com) was used.

TABLE-US-00003 TABLE 3 Comparison GenPept with normal No. Protein name Accession no. (fold) 1 26S proteasome non-ATPase regulatory subunit 13 NP_002808 2.0 isoform 1 2 40S ribosomal protein S12 NP_001007 2.3 3 Apo form of human S100a16 3NXA_A 2.6 4 ASPRV1 protein AAH31997 674.1 5 Bifunctional purine biosynthesis protein PURH NP_004035.2 4.0 6 Cellular retinoic acid binding protein li in complex with 2CBS_A 2140.8 synthetic retinoic acid 7 Aldehyde reductase 2ALR_A 16.9 8 Human epidermal fatty acid-binding protein (fabp5) in 4AZM_A 2.2 complex with the inhibitor Bms-309413 9 Prolyl oligopeptidase with Gsk552 3DDU_A 3.2 10 Solution structure of Apo S100a16 2L50_A 3562.0 11 Structure of [r563a] leukotriene A4 hydrolase 1SQM_A 3.4 12 The high resolution structure of annexin Iii shows 1AXN_A 1935.1 differences with annexin V 13 Three crystal stmctures of human coactosin-like protein 1T2L_A 4.0 14 Cystatin-B NP_000091 2.6 15 Ezrin AAH68458 5.4 16 Isopentenyl-diphosphate delta-isomerase 1; Short = IPPI1 Q13907 3.6 17 Keratin, type I cytoskeletal 17 NP_000413 12.2 18 Macrophage-capping protein isoform 9 XP_515584 4.8 19 Mitochondrial ATP synthase, H+ transporting F1 ABD77240 2.8 complex beta subunit 20 NCOR1 protein, partial AAH58511 2.0 21 Phosphoglycerate mutase 1 (brain) AAH62302 2.4 22 PREDICTED: tubulin alpha-1B chain-like isoform 2 XP_002823231 821.6 23 Protein S100-A8 isoform 4 [Pan troglodytes] XP_001138065 3473.3 24 R33729_1 AAC27824 2.4 25 TALDO1 protein AAH18847 915.0 26 Type I keratin 16 AAB35421 195.4 27 All taxonomy (Crystal Structure of Bovine Serum 3V03_A 5.76 Albumin) 28 Alpha-actinin-4 NP_004915 4.70 29 Alpha-crystallin B chain NP_001876 5.43 30 Crystal structure of human enolase 1 3B97_A 2.00 31 Human peroxiredoxin 5 1HD2_A 12.14 32 Structural and electrophysiological analysis of annexin V 1HVE_A 6.35 mutants. mutagenesis of human annexin V 33 Structure of S100a4 in complex with non-muscle 4ETO_A 13.12 myosin-Iia peptide 34 X-ray crystal structure of human galectin-1 1GZW_B 4.15 35 Structure of the human class I histocompatibility antigen, 1HLA_M 2.18 Hla-A2 36 Chip-Ubc13-Uev1a complex 2C2V_B 4.38 37 Chorionic somatomammotropin hormone-like 1 isoform 3 NP_001309, 2.04 38 Dynactin 3 (p22) CAI13144 3.72 39 Eukaryotic translation initiation factor 2B, subunit 2 AAH00494 7.66 beta, 39 kDa 40 Gelsolin isoform b NP_937895 2.19 41 Glutaredoxin-3 NP_006532 4.21 42 Glutathione S-transferase P NP_000843 3.18 43 Heat shock cognate 71 kDa protein isoform 1 NP_006588 2.41 44 HMOX2 CAG33041 689.62 45 Human Glutathione Transferase Omega 1, Delta 155 3LFL_A 2.94 46 Keratin 1, keratin, type I cytoskeletal 9 AAG41947 950.95 47 LMNB1 protein AAH78178 540.70 48 Moesin ferm domain bound to Ebp50 C-terminal peptide 1SGH_A 3.83 49 Mrp14 complexed with chaps 1IRJ_A 4.07 50 MYO5C protein AAH64841 3.40 51 Nicotinamide N-methyltransferase NP_006160 5.79 52 Nuclear transfactor 2 NP_005787 9.35 53 nucleoside diphosphate kinase A isoform a NP_937818 19.07 54 O-methyltransferase 3BWM_A 3.44 55 Platelet-activating factor acetylhydrolase IB subunit NP_002564 2.40 gamma 56 Protein S100-A11 NP_005611 3.19 57 Pyruvate dehydrogenase (lipoamide) beta EAW65372 5.94 58 Serine/threonine-protein phosphatase 2A catalytic NP_058736 1331.25 subunit beta isoform 59 Tubulin alpha-1B chain isoform 2 XP_002823231 7.94 60 Tumor necrosis factor type 1 receptor associated protein A55877 3.11 TRAP-1-human 61 Ubiquitin carboxyl-terminal hydrolase 14 isoform a NP_005142 4.52 62 Vacuolar protein sorting-associated protein 29 NP_057310 2.52 63 X-ray repair cross-complementing protein 5 NP_066964 7.91

[0098] When the expression of Proteins 1 to 26 shown in Table 3 is increased, the sample can be diagnosed as the skin of a patient with keloid skin or a keloid scar, and when the expression of Proteins 27 to 63 is decreased, the sample can be diagnosed as the skin of a patient with keloid skin or a keloid scar.

TABLE-US-00004 TABLE 4 Comparison GenPept with normal No. Protein name Accession no. (fold) 64 Chain A, structure of S100a4 in complex with non- 4ETO_A 6575.08 muscle myosin-Iia peptide 65 Phosphoglycerate mutase 1 (brain) AAH62302 16.35 66 Putative G-protein coupled receptor BAB89334 7.92 67 Chain A, human quinone reductase type 2 1QR2_A 4.63 68 T-plastin polypeptide AAB02844 10.81 69 FLNA protein AAH14654 5.17 70 Transgelin variant BAD92792 26.98 71 hCG38213, isoform CRA_d EAW76181 3.19 72 Leukotriene A4 hydrolase, isoform CRA_a EAW97557 6.94 73 Prolyl 4-hydroxylase subunit alpha-1 isoform 2 NP_001017962 4.54 precursor 74 Ubiquitin carboxyl-terminal hydrolase isozyme L1 NP_004172 19.98 75 Rho GDP-dissociation inhibitor 1 isoform a NP_004300 2.75 76 Cytokeratin-14 P02533 2051.74 77 Keratin 5 AAH24292 1291.44 78 Prohibitin AAS88903 2.21 79 FLJ00410 protein BAC03467 2.32 80 Serpin B5 NP_002630 7150.72

[0099] When the expression of Proteins 64 to 75 shown in Table 4 is increased, the sample can be diagnosed as the skin of a patient with keloid skin or a keloid scar, and when the expression of Proteins 76 to 80 is decreased, the sample can be diagnosed as the skin of a patient with keloid skin or a keloid scar.