JP2007163407A - Post-operative prognosis testing method for early-stage lung cancer - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide prognosis testing method, etc. for patients with lung cancer which clarifies the work of esRAGE in vivo, while utilizing its measurements. <P>SOLUTION: This post-operative prognosis testing method for the patients with lung cancer, which includes measuring of the amount of manifestation endogenous secretory RAGE (Endogenous Secretory Receptor for Advanced Glycation End products: esRAGE), a test kit used for the above method, a prognosis testing reagent for the patients with lung cancer comprising anti-esRAGE antibody, etc. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a postoperative prognostic test method for lung cancer patients, comprising measuring the expression level of endogenous secretory receptor for advanced glycation end products (esRAGE), a test kit used therefor, and an anti-esRAGE antibody The present invention relates to a reagent for prognosis examination of lung cancer patients.

  The number of lung cancer cases is increasing worldwide. For lung cancer without distant metastases, surgical treatment is the best prognosis. However, even in early stage (Stage I) lung cancer, the 5-year survival rate remains 65% after the operation, and the remaining 35% is relapsed or recurring despite curative resection. He died from distant metastasis (References 1-3). These poor prognosis cases cannot be distinguished at the stage of completion of surgery, and require a long follow-up period thereafter.

  Therefore, if it becomes possible to judge this poor prognosis case early after surgery, it becomes possible to detect recurrence early by additional treatment (chemotherapy with anticancer drugs) and careful follow-up. It is considered possible to prolong the prognosis. At the same time, unnecessary examinations and treatments can be reduced for cases with good prognosis, leading to a reduction in medical costs. Therefore, it is important to develop predictive factors for postoperative prognosis of early lung cancer.

  Receptor for AGE (RAGE) is a receptor that has various substances as ligands and is involved in inflammation, diabetic angiopathy, cancer metastasis, etc. (Advanced Glycation Endproducts: AGE) References 4, 5). In particular, it was revealed that RAGE and its ligand HMGB-1 / amphoterin are greatly involved in cancer invasion and metastasis (Reference Documents 6, 7, and 8). Recently, it has been discovered that this RAGE has an isoform called endogenous secreted RAGE (esRAGE) produced by alternative splicing from one gene (Reference 9, Patent Document 1, Patent Document 2).

It is thought that esRAGE binds to the ligand of RAGE and competes with RAGE (full-length RAGE) as an original receptor to suppress the cell response via RAGE. However, its function in vivo is not clear. Patent Literature 1 and Patent Literature 2 describe the possibility that nucleic acids encoding them and anti-esRAGE antibodies can be used for the treatment or prevention of various diseases such as diabetic complications, methods for measuring esRAGE using esRAGE antibodies, and The susceptibility and resistance predictability of diabetic complications by quantifying esRAGE are described. Patent Document 3 describes an invention relating to a method for prognosing lung cancer, but what is actually being studied relates to patients in the later stages of clinical stage III and IV.
JP 2003-125786 A JP 2003-128700 A Japanese Patent Laid-Open No. 11-230966

  Accordingly, an object of the present invention is to clarify the function of esRAGE in vivo and to provide a prognostic method for lung cancer patients using the measurement.

  In order to solve the above-mentioned problems, the present inventor examined the expression of esRAGE in cancer tissues and healthy tissues using 182 non-small cell lung cancer surgical cases and clarified the relationship with prognosis, thereby completing the present invention.

That is, the present invention relates to the following aspects.
[1] A method for prognosing lung cancer patients, comprising measuring the expression level of endogenous secretory RAGE (esRAGE) in a specimen sample derived from a lung cancer patient.
[2] The examination method according to aspect 1, wherein the lung cancer patient is in clinical stage I.
[3] The examination method according to aspect 1 or 2, wherein a lung tissue section removed after surgery is used as a specimen sample.
[4] The examination method according to aspect 1 or 2, wherein a preoperative bronchoscopic biopsy specimen is used as a specimen sample.
[5] The test method according to any one of aspects 1 to 4, wherein the expression level of esRAGE is measured by an antigen-antibody reaction using an anti-esRAGE antibody.
[6] The test method according to aspect 5, wherein a labeled antibody is used.
[7] The examination method according to any one of aspects 5 and 6, wherein the expression level of esRAGE is measured by an immunostaining method.
[8] A reagent for prognosis testing of lung cancer patients, comprising an anti-esRAGE antibody.
[9] The reagent according to embodiment 8, which is a labeled antibody.
[10] An inspection kit for use in the inspection method according to any one of aspects 1 to 7, comprising the reagent according to any one of aspects 8 or 9.
[11] An anti-lung cancer agent containing a nucleic acid molecule encoding esRAGE as an active ingredient.
[12] The anti-lung cancer agent according to 11, wherein the nucleic acid molecule is incorporated into a vector.
[13] An anti-lung cancer agent containing esRAGE as an active ingredient.

  Conventionally, there was no factor that predicts the prognosis of postoperative patients with early lung cancer. According to the present invention, it has been clarified that the decrease in esRAGE expression of cancer cells correlates with the prognosis of the case. This correlation was particularly observed in early lung cancer cases. Therefore, by measuring esRAGE expression in lung cancer surgical tissue, it becomes possible to examine and determine the prognosis of lung cancer patients, especially early lung cancer patients, without looking at the long-term progress based on the expression level. It was.

  The endogenous secreted RAGE (esRAGE) to be measured in the prognostic test method of the present invention usually means a human-derived protein that is actively produced and secreted in vivo among soluble RAGEs. The substance is described in detail in, for example, Patent Document 1 or Patent Document 2 (however, labeled “soluble RAGE”), and is a substance known to those skilled in the art.

  That is, more specifically, “endogenous secreted RAGE (esRAGE)” is a peptide related to receptor for advanced glycation endproducts (RAGE), which is closely related to diabetic complications, and is a splicing variant of RAGE. It refers to a peptide that does not have a penetrating region.

The esRAGE is a peptide consisting of 347 amino acid residues (however, if it is actually secreted outside the cell, the signal sequence consisting of 22 amino acid residues is removed and the peptide consisting of 325 amino acid residues) And a characteristic sequence on the C-terminal side: GluGlyPheAspLysValArgGluAlaGluAspSerProGlnHisMet,
And is characterized by lacking a transmembrane domain present in transmembrane RAGE (also referred to as membrane-type RAGE or membrane-bound RAGE). The esRAGE has an advanced glycation endproducts (AGE) binding activity, or has an inhibitory or inhibitory activity on the interaction between AGE and its receptor. Typically, the esRAGE of the present invention is a naturally occurring peptide (endogenous peptide or endogenous peptide) that exists in a living body and differs from the RAGE protein in 16 amino acid residues in the C-terminal part. . A typical soluble RAGE of the present invention is a polypeptide encoded and produced by the DNA of SEQ ID NO: 1 of Sequence Listing described in Patent Document 1 or 2, for example, the amino acid of SEQ ID NO: 2 of Sequence Listing A polypeptide having the sequence or a substantially equivalent amino acid sequence is mentioned. The typical soluble RAGE of the present invention has at least 1 to 16 consecutive amino acid residues in the C-terminal side of the amino acid sequence Glu ^{332 to} Met ³⁴⁷ of SEQ ID NO: 2 in the sequence listing. And having AGE binding activity, having at least ^{1 to 117} consecutive amino acid residues on the N-terminal side of amino acid sequence Met ^{1 to} Val ¹¹⁷ of SEQ ID NO: 2 in the sequence listing and having AGE binding activity Or those having these characteristics and having at least 60% homology with the amino acid sequence Tyr ^{118 to} Gly ³³¹ of SEQ ID NO: 2 in the Sequence Listing.

  Here, “homology” refers to each amino acid residue or each base constituting the chain between two chains in a polypeptide sequence (or amino acid sequence) or polynucleotide sequence (or base sequence). It means the amount (number) of things that can be determined to be identical in the mutual relationship of each other, and means the degree of sequence correlation between two polypeptide sequences or two polynucleotide sequences. Homology can be easily calculated. Many methods for measuring homology between two polynucleotide or polypeptide sequences are known, and the term “homology” (also referred to as “identity”) is well known to those skilled in the art (eg, , Lesk, AM (Ed.), Computational Molecular Biology, Oxford University Press, New York, (1988); Smith, DW (Ed.), Biocomputing: Informatics and Genome Projects, Academic Press, New York, (1993); Grifin , AM & Grifin, HG (Ed.), Computer Analysis of Sequence Data: Part I, Human Press, New Jersey, (1994); von Heinje, G., Sequence Analysis in Molecular Biology, Academic Press, New York, (1987 Gribskov, M. & Devereux, J. (Ed.), Sequence Analysis Primer, M-Stockton Press, New York, (1991), etc.). Common methods used to measure the homology of two sequences include Martin, J. Bishop (Ed.), Guide to Huge Computers, Academic Press, San Diego, (1994); Carillo, H. & Lipman , D., SIAM J. Applied Math., 48: 1073 (1988), etc., but are not limited thereto. Preferred methods for measuring homology include those designed to obtain the largest match between the two sequences being tested. An example of such a method is one assembled as a computer program. Preferred computer programming methods for measuring homology between two sequences include the GCG program package (Devereux, J. et al., Nucleic Acids Research, 12 (1): 387 (1984)), BLASTP, BLASTN, FASTA (Atschul, SF et al., J. Molec. Biol., 215: 403 (1990)) and the like, but are not limited thereto, and methods known in the art can be used. .

  As a specimen sample, any part of lung tissue extracted from a lung cancer patient can be used. As an example of a suitable specimen sample, for example, a lung tissue section removed after surgery, or a bronchoscope before surgery List biopsy specimens. These specimen samples are subjected to an appropriate method according to the measurement method and the like and then used for the inspection method. For example, it is possible to use a sample extracted and prepared from the tissue specimen or the like by an appropriate means known to those skilled in the art.

  There is no particular limitation on the clinical stage of lung cancer patients to be examined, but it is more significant when specimen samples from patients with early lung cancer such as clinical stage (TNM stage) stage I are used. Prognostic test results can be obtained, which is advantageous.

  The expression level of esRAGE can be measured by any method known to those skilled in the art. Examples include, for example, a method using various immunological specific reactions such as immunostaining using an anti-esRAGE antibody and EIA, an amino acid sequence analysis method for peptides such as a gas phase sequencer using Edman method, and MALDI- It can detect by mass spectrometry represented by TOF / MS, ESI Q-TOF / MS method, etc.

  Among the above methods, a test method for measuring the expression level of esRAGE by an antigen-antibody reaction using an anti-esRAGE antibody is preferable. The antibody can be prepared by an appropriate method known to those skilled in the art using the above esRAGE or an appropriate partial polypeptide (peptide fragment) thereof or various derivatives or complexes thereof as an antigenic substance or an immunogen. Is possible. For example, in the case of a polyclonal antibody, it can be administered to an appropriate animal such as a mouse, rat, rabbit, goat or chicken and prepared from the antiserum. Alternatively, a known monoclonal antibody production method (“monoclonal antibody”, Kamei Nagamune, Hiroaki Terada, Yodogawa Shoten, 1990; “Monoclonal Antibody” James W. Goding, third edition, Academic Press, 1996), etc. It is also possible to prepare a monoclonal antibody by a method using cell fusion. For example, it can be prepared by the method described in Patent Document 1 or Patent Document 2. Specific examples of the anti-esRAGE antibody prepared by such a method include anti-human esRAGE antibodies listed in Table 1 of the same patent document.

Such an anti-esRAGE antibody is derived from a complete antibody such as Fab, F (ab ′) ₂ , Fv fragment, etc. by genetic engineering (DNA recombination technology) unless the original antibody activity is lost. It may be a derivative, recombinant or fragment modified into various forms known to those skilled in the art, including various derivatives of

  In addition, as described in the Examples of the present specification, esRAGE itself is prepared by culturing various cell lines transformed with a gene encoding the same, and producing esRAGE in the transformed cells. Alternatively, it can be produced by a chemical synthesis method.

  The gene encoding esRAGE is a template of an appropriate cDNA library prepared on the basis of various human tissues such as primary cultured human skin microvascular endothelial cells or human brain tissue. Known to those skilled in the art such as PCR using primers designed appropriately based on the nucleotide sequence, and other NASBA (Nucleic acid sequence based amplification), TMA (Transcription-mediated amplification) and SDA (Strand Displacement Amplification) methods By using an arbitrary DNA amplification technique, it can be easily obtained as cDNA of the gene.

  Alternatively, the gene can be isolated by screening the cDNA library by methods well known to those skilled in the art. Furthermore, the cDNA of the gene can be prepared by introducing a base mutation using a commercially available mutation system or the like based on site-directed mutagenesis known to those skilled in the art.

  In addition, the above gene can be obtained by a known method (for example, Carruthers (1982) Cold Spring Harbor Symp. Quant. Biol. 47: 411-418; Adams (1983) J. Am. Chem. Soc. 105: 661; Belousov (1997). ) Nucleic Acid Res. 25: 3440-3444; Frenkel (1995) Free Radic. Biol. Med. 19: 373-380; Blommers (1994) Biochemistry 33: 7886-7896; Narang (1979) Meth. Enzymol. 68:90 Synthesized in vitro by well-known chemical synthesis techniques such as described in Brown (1979) Meth. Enzymol. 68: 109; Beaucage (1981) Tetra. Lett. 22: 1859; US Pat. No. 4,458,066) You can also. It can also be produced by a method such as cleaving the polynucleotide of the present invention with an appropriate restriction enzyme.

  Thus, the prognosis of the lung cancer patient can be examined by measuring the expression level of esRAGE in the specimen sample derived from the lung cancer patient using the anti-esRAGE antibody. In addition, the measurement of the expression level is not necessarily quantitative, and the effect of the present invention can be sufficiently obtained even by qualitative or semi-quantitative determination by visual observation according to a specific measurement method / means. I can do it. Accordingly, the anti-esRAGE antibody has a use as a prognostic test reagent for lung cancer patients.

  In the prognosis test method for lung cancer patients of the present invention, the expression level of esRAGE in a specimen sample derived from a lung cancer patient can also be measured by the amount of mRNA encoding it.

  As described in Examples in the present specification, such mRNA measurement is performed, for example, by RT-PCR method using primers appropriately designed based on the nucleotide sequence of the gene described in Patent Document 1 and the like These methods can be performed by methods known to those skilled in the art, such as various quantitative PCR methods and microarray (DNA chip) methods.

  Furthermore, a test kit including a prognostic test reagent as described above, for example, used in a prognostic test method for lung cancer patients can have an appropriate configuration according to the specific measurement principle of esRAGE. The kit can contain, for example, a prognostic test reagent comprising an anti-esRAGE antibody, or a primer for amplification of the esRAGE gene and a probe for hybridization for the measurement of the above mRNA. These consist of a continuous base sequence of an appropriate length, for example, 10 to 100, depending on the application.

  In the chemical synthesis of esRAGE polypeptide and the preparation of its gene, the design of various primers or probes described so far, the amino acid sequence information of esRAGE described in Patent Document 1 or Patent Document 2 and the base of the gene Sequence information can be consulted.

  Various primers, probes, or antibodies included as components in the above kit may be labeled with an appropriate labeling substance such as any radioactive substance, fluorescent substance, and dye known to those skilled in the art. Further, the kit includes other elements or components known to those skilled in the art, such as various reagents, enzymes, buffers, reaction plates (containers), and the like, depending on the configuration and purpose of use.

  The anti-lung cancer agent of the present invention contains a nucleic acid molecule encoding esRAGE or esRAGE itself as an active ingredient. As shown in the examples of the present specification, by introducing a nucleic acid molecule encoding esRAGE into cancer cells and forcibly expressing esRAGE, or administering an anti-lung cancer agent containing esRAGE as an active ingredient By causing the esRAGE active ingredient to act on cancer cells, the growth of cancer cells can be suppressed.

  Nucleic acid molecules encoding esRAGE are inserted into appropriate vectors used for gene therapy such as various viral vectors such as retroviral vectors, adenoviral vectors, and adeno-associated viral vectors, non-viral vectors, or hybrid vectors. It is preferable to use it. Such vectors appropriately contain various elements such as various gene expression regulatory sequences, cloning sites, drug resistance genes and the like, and can be prepared by any method known to those skilled in the art.

  The anti-lung cancer agent may contain other components such as any pharmaceutical carrier or diluent known to those skilled in the art, in combination with an active ingredient, which are pharmaceutically acceptable.

  The pharmaceutically effective amount and administration method or means of administration of the active ingredient of the present invention include the type of active ingredient, the severity of the disease state, the treatment policy, the patient's age, weight, gender, general health condition, and the patient's ( It can be appropriately selected by those skilled in the art depending on the genetic background. For example, when esRAGE itself is contained as an active ingredient, the dose of the active ingredient is 0.1 to 100 mg / day / adult, more generally 1 to 10 mg / day / adult.

  The anti-lung cancer agent of the present invention can have any shape known to those skilled in the art depending on the administration method, administration route and the like. They can be administered by any suitable method. For example, liquids, powders, colloids, etc. can be used, and can be injected intravenously, intraperitoneally, subcutaneously, or orally with the carrier or diluent described above. Can be mentioned.

  EXAMPLES Hereinafter, although an Example demonstrates this invention in detail, the technical scope of this invention is limited and is not interpreted at all by description of a following example. Also, unless otherwise noted, the following examples are routine methods in the art and standard methods known to those skilled in the art, such as Sambrook and Maniatis, in Molecular Cloning-A Laboratory Manual, Cold Spring Harbor. Laboratory press, New York, 1989; Ausubel, FM et al., Current Protocols in Molecular Biology, John Wiley & Sons, New York, NY, 1995. Moreover, the description content of the literature cited as a reference etc. in this specification constitutes a part of the disclosure content of this specification.

Case
182 non-small cell lung cancer patients who underwent surgery at Sendai Kosei Hospital from 1993 to 1995 and were not treated with postoperative chemotherapy or radiotherapy were included. The average age of the cases was 65.3 years (23-82 years). In 36 cases, frozen tissues were obtained and used for RNA analysis. This study is accredited by the Ethics Committee of Tohoku University and Sendai Kosei Hospital.

Statistical analysis Statistical analysis of various results obtained in this example was performed using “a one-way ANOVA and Bonferroni test” or χ 2 test. Overall survival curves and non-disease survival curves were generated according to the Kaplan-Meier method. Univariate analysis and multivariate analysis were performed with Cox's proportional hazard model using PROC PHREG, and Fisher's direct method was implemented with SAS software. In this test, a P value of less than 0.05 was determined to be significant.

The immunostained lung was fixed with 10% formalin, embedded in paraffin, sliced, and stained with anti-human esRAGE antibody. In addition, staining was performed using anti-Ki-67 antibody to see the proliferation of cancer cells. To see the specificity of esRAGE staining, an absorption test was performed using lycobinnant esRAGE protein.
The anti-human esRAGE antibody first immunizes rabbits by chemically synthesizing a peptide with 16 amino acids (EGFDKVREAEDSPQHM) characteristic of human esRAGE with Cys added to the N-terminal side in the usual manner. It was prepared by. In addition, the recombinant esRAGE protein is forcibly expressed in cultured COS-7 cells and 293 cells by expressing an expression vector (pCI-neo-mammalian expression vector, Promega) in which human esRAGE cDNA has been incorporated (described later). Obtained by purifying from a clean to high purity.

esRAGE is expressed in the cytoplasm of normal airway epithelial cells and stromal cells (FIGS. 1A and B), which is consistent with the previously reported result (Reference 10). However, esRAGE expression varies among cancer cells. As a result of visual assessment, esRAGE positive cases (+) of cancer cells are 45 cases (24.7%), and decreased expression cases (+/-) are 78 cases. There were 59 cases (32.4%) and 4 cases (42.9%) and negative cases (-). In cases with high pathological factor T (P = 0.0063), histologically poorly differentiated (P = 0.0061), and proliferating (P <0.0001), there were many cases of decreased esRAGE expression or negative cases (Table 1). ). Table 1 shows the correlation with esRAGE immunostaining and clinicopathological parameters in 182 non-small cell lung cancer (NSCLCs) cases.

  Furthermore, examination of the correlation between esRAGE expression and survival rate revealed that the survival rate was low in cases where esRAGE expression fell in cancer cells (P = 0.0003) (Fig. 2A). This tendency was particularly prominent in patients with early lung cancer at TNM Stage I (P = 0.0001) (Fig. 2B). Univariate analysis showed that TNM staging (P <0.0001) and esRAGE staining of cancer cells (-, +/- vs. +) (P <0.001) were prognostic factors, and multivariate analysis TNM staging (P <0.0001) and cancer cell esRAGE staining (P = 0.0085) were found to be independent prognostic factors (Table 2). Table 2 shows the results of univariate and multivariate analysis on the clinical results of 182 non-small cell lung cancer (NSCLCs) cases.

EsRAGE mRNA expression in cancer cells To examine esRAGE mRNA expression in cancer cells, the cancer cells and peripheral cells in non-small cell lung cancer (NSCLCs) cases were separated from frozen cancer tissue by laser capture method, and total RNA from each Were extracted, and the expression of esRAGE mRNA and full-length RAGE was analyzed by RT-PCR. Β-actin was used as an endogenous control. The primer sequences used were human esRAGE, 5'-TCT GTG GGG GGC AGT AGT A-3 '(SEQ ID NO: 1) and 5'-CTT TAT CAA ACC CCT CAC CT-3' (SEQ ID NO: 2); full length RAGE, 5'-TCT GTG GGG GGC AGT AGT A-3 '(SEQ ID NO: 3) and 5'-TGC CGC CTT TGC CAC AAG AT-3' (SEQ ID NO: 4); human beta-actin, 5'-GCT GTG CTA TCC CTG TAC G-3 ′ (SEQ ID NO: 5) and 5′-TGC CTC AGG GCA GCG GAA-3 ′ (SEQ ID NO: 6).

  As a result, esRAGE or full-length RAGE expression was lost in 25 out of 36 cases (70%) only in cancer cells (Table 3A). However, there was no correlation between esRAGE and full-length RAGE mRNAs expression (p = 0.72, Table 3B).

Effect of esRAGE on cancer cell proliferation
In order to examine the effects of esRAGE on cancer cell growth, we introduced esRAGE, full-length RAGE or dominant negative RAGE (dnRAGE) cDNA with promoters in lung cancer cells A549 and OBA-LK-1 cells. The effect of forced expression on cell proliferation was examined. That is, human esRAGE, full-length RAGE, dominant negative RAGE, or an empty vector was introduced into 5 × 10 ⁵ A549 or OBA-LK-1 cells using the liposome method. Proliferation after the introduction was measured by adding fluorescence of alamarBlue (Biosource, Camarillo, CA) to cultured cells and measuring fluorescence intensity at 24, 48, 72, and 96 hours after culturing.
Here, esRAGE and full-length RAGE cDNA were cloned from cultured vascular cells, and dnRAGE was artificially prepared by genetic manipulation (PCR) in which a stop codon was inserted immediately after the transmembrane region sequence of full-length RAGE. They were prepared by incorporating them into pCI-neo mammalian expression vector (Promega), respectively. The promoter of this vector uses the human cytomegalovirus immediate-early enhancer / promoter region.
Specifically, full-length RAGE cDNA is 5′-GAGAATTCGCCAGGACCCTGGAAGGAAGCA-3 ′ (SEQ ID NO: 7) corresponding to nucleotides 1-22 and 1246-1268 of AB036432 (Genbank accession number), respectively. 5'-GATCTAGACTGATGGATGGGATCTGTCTGTG-3 '(SEQ ID NO: 8), esRAGE is 5'-GAGAATTCGCCAGGACCCTGGAAGGAAGCA-3' (SEQ ID NO: corresponding to nucleotides 1-22 and 1069-1088 of AB061668 (Genbank accession number), respectively. 9) and 5′-GATCTAGAGCTTGTTGACCATCCCCCCAG-3 ′ (SEQ ID NO: 10) were subjected to RT-PCR, and dnRAGE was 5′-GAGAATTCGCCAGGACCCTGGAAGGAAGCA-3 ′ (SEQ ID NO: 11) and 5′-GATCTAGATCATCGGCGTTGCCGCCTTTG-3 ′ (SEQ ID NO: 12) PCR was carried out in, and each sequence was confirmed. Then, each fragment was digested with restriction enzymes EcoRI and XbaI and incorporated into pCI-neo mammalian expression vector (Promega) to prepare each expression vector. Lung cancer cells A549 (ID: TKG 0184) and OBA-LK-1 cells (ID: TKG 0572) are both stored at the Medical Cell Resource Center, Institute for Aging Medicine, Tohoku University and can be freely distributed. .

As a result, suppression of cancer cell proliferation was observed by forcibly expressing esRAGE in cancer cells (FIG. 3). This suggests that the growth of cancer is promoted by the decrease of esRAGE expression in lung cancer cases.

[References]
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3. Spiro SG, Porter JC. Lung Cancer--Where Are We Today? Current Advances in Staging and Nonsurgical Treatment. Am J Respir Crit Care Med 2002; 166 (9): 1166-96.
4. Neeper M, Schmidt AM, Brett J, et al. Cloning and expression of a cell surface receptor for advanced glycosylation end products of proteins.Journal of Biological Chemistry 1992; 267 (21): 14998-5004.
5. Schmidt AM, Yan SD, Yan SF, Stern DM. The multiligand receptor RAGE as a progression factor amplifying immune and inflammatory responses. J Clin Invest 2001; 108 (7): 949-55.
6. Sakurai S, Yonekura H, Yamamoto Y, et al. The AGE-RAGE system and diabetic nephropathy.J Am Soc Nephrol 2003; 14 (8 Suppl 3): S259-63.
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8. Huttunen HJ, Fages C, Kuja-Panula J, Ridley AJ, Rauvala H. Receptor for advanced glycation end products-binding COOH-terminal motif of amphoterin inhibits invasive migration and metastasis. Cancer Res 2002; 62 (16): 4805- 11.
9. Yonekura H, Yamamoto Y, Sakurai S, et al. Novel splice variants of the receptor for advanced glycation end-products expressed in human vascular endothelial cells and pericytes, and their putative roles in diabetes-induced vascular injury. Biochem J 2003; 370 (Pt 3): 1097-109.
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  According to the present invention, it becomes possible to identify early lung cancer patients with poor prognosis immediately after the operation, and it is considered that the prognosis can be improved by performing early additional treatment (anticancer drug treatment) on these patients. On the other hand, if a case with a good prognosis is identified, unnecessary postoperative therapy and excessive postoperative examinations / outpatient examinations can be omitted, leading to a reduction in medical costs.

In the present invention, the expression of esRAGE can be measured not only after surgery but also with a preoperative bronchoscopic biopsy specimen. This leads to the evaluation of the prognosis after surgery before surgery and the development of new treatment strategies for such cases.

It is a microscope picture which shows the expression of esRAGE in a non-small cell lung cancer tissue. A: Lung cancer tissue, B: Healthy lung tissue, C: esRAGE absorption test. Bar scale = 100 μm. The postoperative survival curve of a lung cancer operation case is shown. A: All cases, B: TNM stage I cases, C: TNM stage II and IIIA cases. It shows that the growth of A549 cultured lung cancer cells (A) and OBA-LK-1 (B) cultured lung cancer cells is suppressed by overexpression of esRAGE. * P <0.05 (vs. empty vector); † P <0.05 (vs. full-length RAGE).

Claims (13)

A method for prognosing lung cancer patients, comprising measuring the expression level of endogenous secretory RAGE (esRAGE) in a specimen sample derived from a lung cancer patient. The test method according to claim 1, wherein the lung cancer patient is in clinical stage I. The examination method according to claim 1 or 2, wherein a lung tissue section removed after surgery is used as a specimen sample. The examination method according to claim 1 or 2, wherein a preoperative bronchoscopic biopsy specimen is used as a specimen sample. The test | inspection method as described in any one of Claims 1-4 which measures the expression level of esRAGE by the antigen antibody reaction which uses an anti- esRAGE antibody. The test method according to claim 5, wherein a labeled antibody is used. The test | inspection method as described in any one of Claim 5 or 6 which measures the expression level of esRAGE with an immuno-staining method. A reagent for prognosis testing of lung cancer patients, comprising an anti-esRAGE antibody. The reagent according to claim 8, which is a labeled antibody. A test kit for use in the test method according to any one of claims 1 to 7, comprising the reagent according to any one of claims 8 or 9. An anti-lung cancer agent containing a nucleic acid molecule encoding esRAGE as an active ingredient. The anti-lung cancer agent according to claim 11, wherein the nucleic acid molecule is incorporated into a vector. An anti-lung cancer agent containing esRAGE as an active ingredient.

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