JP5526378B2 - Method for detecting cervical cancer - Google Patents

Description

  The present invention relates to a method for detecting cancer peculiar to women, specifically cervical cancer, more specifically cervical squamous cell carcinoma, specifically, a protein expressed in cancer tissue and non-cancer tissue. Markers of cervical squamous cell carcinoma consisting of a protein whose expression is specifically enhanced or decreased, and a method for detecting cancer using the same and screening for cancer therapeutics, revealed by comprehensive expression analysis of Regarding the method.

  Cervical cancer is a cancer that occurs at the exit of the uterus, called the cervix, and corresponds to many “uterine cancers” that occur in the uterus. Most cervical cancers are squamous cell carcinomas that occur in the cervix, and other types include mucinous adenocarcinoma and mucoepidermoid carcinoma. Among these, most cervical squamous cell carcinomas are caused by persistent infection with human papillomavirus (HPV), and there is a characteristic that there is almost no subjective symptom in the early stage of onset.

  In cancer countermeasures, surgical operations and medical treatments are tried, but the most important thing is to find them early and select appropriate treatments. From this point of view, cervical cytology has been conventionally used for screening for cervical cancer. This is a traditional test method in which cells collected from the uterine cervix are stained with pigments and microscopically observed for abnormal cells, and is an effective method for detecting cancer or precancerous lesions. Currently, the screening rate for uterine cancer screening in Japan is around 20%, which is low in developed countries, and the screening rate for young people (HPV is infected by sexual intercourse is higher than that for other cancers) There was a big problem of being sluggish. In fact, it has been pointed out that the onset of cervical cancer has become younger, and an increase in the number of affected individuals has been pointed out (Non-Patent Documents 1 and 2). In order to increase the screening rate and lead to early detection, it is desirable to develop and popularize a more acceptable screening, for example, measurement of tumor markers by blood sampling.

  For example, in prostate cancer, a prostate-specific antigen (PSA) has been discovered as a tumor marker that can be measured from blood, and by introducing this marker test, the detection rate of early cancer will be increased to 60% or more. connected. In addition, protein disulfide isomerase (PDI) is known as a tumor marker for prostate cancer (Patent Document 1). In cervical cancer, several types of conventionally used tumor markers, such as SCC and CYFRA, are known, but they are useful but useful for monitoring therapeutic effects in cervical cancer that has already progressed or as a measure of recurrence of cervical cancer. The positive rate in cervical cancer is not sufficient, and the present condition is that the early detection means of cervical cancer has not been established (nonpatent literatures 3 and 4).

  In addition to surgery or radiation therapy, chemotherapy has been adopted as a treatment method for cervical cancer, but so far, chemotherapy has not been effective enough for cervical cancer, especially advanced cervical cancer and recurrence. The prognosis for cervical cancer was very poor (Non-patent Document 5). In recent years, molecular targeted therapeutic drugs have appeared in various types of cancer, and the therapeutic methods are changing drastically. Since molecular targeted therapeutic drugs target molecules that are specifically expressed in cancer, it is expected that tumor growth can be effectively suppressed with a small dose and side effects. The key to developing such drugs is to perform comprehensive protein expression analysis in cancer tissues / non-cancer tissues and identify molecules that are targets for drug discovery. Comparison of cancer tissue / non-cancerous tissue and search for biomarkers using proteomics in various cancers are one of the methods that are now widely used. Usually, such a comparison is performed in the same individual, and a biomarker search is performed by identifying proteins or the like whose expression is specifically increased or decreased in cancer.

However, in cervical cancer, in many cases, the lesion occupies the entire cervix, and there is a problem that non-cancerous tissue is rarely left. In recent years, the number of radiation therapy cases has increased, and the number of surgical specimens has been reduced, making it difficult to compare the same individuals using the proteomic method. In order to solve this problem, Gu Y. et al. Et al. Have separated normal cells and tumor cells from slides that have undergone cytodiagnosis, and examined the expression of proteins in each of them (Non-patent Document 6). Although this method is evaluated in terms of using specimens from the same person, normal and tumor cells are discriminated only by morphological changes, and the characteristics of tissues in vivo and biological responses to tumors are determined. I missed it. Ono A.I. (Non-patent Document 7) conducts proteomic analysis using preserved specimens fixed in formalin and embedded in paraffin, but normal tissue and cancer tissue were collected from different patients, There is a problem that the influence cannot be ignored because of the strong pretreatment, and a possibility that the biological response to the tumor is leaked or modified during the tissue fixation process.
Moreover, as a marker of uterine cancer, nuclear matrix protein, cytokeratin, Nup358, lamin A expression variation (Patent Document 2), mass 4769 ± 3 (m / z), 6254 ± 4 (m / z) or 11792 ± 6 ( m / z) (PTL 3), histamine release factor (HRF) protein expression variation (PTL 4) and HPV-16 genomic DNA CpG methylation A method for detecting uterine cancer as an index (Patent Document 5) and the like have been disclosed so far, but none of them has been said to be sufficiently effective.
Thus, for cervical cancer, the development of a marker that can detect cervical cancer with high accuracy and high efficiency has not been established since screening methods such as effective tumor markers and therapeutic agents using the same have not been established. Was desired.

  In view of the above current situation, the present invention is a biomarker protein that can detect cancer specific to women, particularly cervical cancer with high accuracy, and can be used as a tumor marker or drug discovery target, and cervical cancer using the same. It is an object of the present invention to provide a detection method and a screening method for a cervical cancer therapeutic agent.

In order to solve the above problems, the present inventors tried to detect a protein specifically expressed in cervical cancer by comprehensive analysis using a proteomic method. When using this method, it is necessary to exclude individual differences and to ensure a sufficient number of cases in terms of statistics. As described above, in cervical cancer, it is very difficult to obtain both non-cancerous tissue and cancerous tissue from the same person. We analyzed protein expression in non-cancerous tissue. As a result, since there was no significant difference in protein expression in these tissues, cancer tissues and non-cancerous tissues were compared in the next step, and proteins with different expression were identified. In order to further increase the number of cases and to examine the difference in the expression level depending on the progression of cervical cancer, 10 normal cases (non-cancerous specimens), 10 cervical cancers (early cervical cancer 5 cases, advanced cervical cancer) 5 cases of cancer) were added, and the expression level of the protein previously identified using the Western blotting method was examined in detail.
Through the above procedure, the present inventors ensure not only tumor cells as in the past but also the whole tumor tissue and a fresh tissue without pretreatment in a statistically sufficient case, and exclude individual differences. Thus, the inventors have found proteins whose expression varies specifically for cervical cancer, that is, crystallin, transgelin, and protein disulfide isomerase (PDI). The present inventors are proteins whose expression is significantly increased or decreased in cervical cancer regardless of individual differences, and by using this as a marker, cervical cancer is efficiently detected. We have found that we can do it and have completed the present invention.

That is, the first aspect of the present invention provides a method for detecting cervical cancer, characterized by detecting expression attenuation of alpha crystallin B chain in crystallin in a biological sample.

The second aspect of the present invention is to detect a decrease in the expression of alpha crystallin B chain in a biological sample and an increase in expression of transgelin and / or protein disulfide isomerase (hereinafter also referred to as PDI). A method for detecting cervical cancer according to the first aspect is provided.

The third aspect of the present invention is an antibody obtained by using the polypeptide of SEQ ID NO: 2 or a polypeptide having 95% or more homology with SEQ ID NO: 2 in the detection of alpha crystallin B chain , and / or PDI Detection of cervical cancer according to the first or second aspect using an antibody obtained by using the polypeptide of SEQ ID NO: 3 or a polypeptide having a homology of 95% or more with SEQ ID NO: 3 in the detection of Provide a method.

The fourth aspect of the present invention is the detection of transgelin, an antibody obtained by using the polypeptide of SEQ ID NO: 2 or a polypeptide having 95% or more homology with SEQ ID NO: 2 and / or detection of PDI The method for detecting cervical cancer according to the third aspect is provided using an antibody obtained by using the polypeptide described in SEQ ID NO: 3 or a polypeptide having 95% or more homology with SEQ ID NO: 3 as an antigen.

  According to a fifth aspect of the present invention, there is provided a kit for detecting or diagnosing cervical cancer for carrying out the method according to the first or second aspect.

  According to a sixth aspect of the present invention, there is provided a kit for detecting or diagnosing cervical cancer comprising the antibody according to the third or fourth aspect.

The seventh aspect of the present invention provides a screening method for a cervical cancer therapeutic agent using an increase in alpha crystallin B chain expression in a test cell as an index.

The eighth aspect of the present invention provides a method for screening a therapeutic agent for cervical cancer using as an index the increase of alpha crystallin B chain expression in a test cell and the attenuation of transgelin and / or protein disulfide isomerase (PDI) expression. To do.

  By utilizing the present invention, cervical cancer can be efficiently detected. In addition, by using the screening method provided by the present invention, it becomes possible to search for a novel therapeutic agent for cervical cancer.

An example of the two-dimensional electrophoresis image of the protein expressed in N group is shown. An example of the two-dimensional electrophoresis image of the protein expressed in C group is shown. The detection result of the crystallin expression in cervical cancer and its surrounding tissue using an anti-crystallin antibody is shown.

  The form for implementing this invention is shown below. According to a first aspect of the present invention, there is provided a method for detecting cancer specific to women, particularly cervical cancer, characterized by detecting an attenuation of crystallin expression in a biological sample. Crystallin is a type of structural protein that constitutes the lens of the eyeball, and three types of α, β, and γ are known. It exists in skeletal muscle, cardiac muscle, kidney, etc., and some crystallins are also known for enzyme activity. In addition, α-crystallin has been pointed out to be similar in primary structure to a low molecular weight heat shock protein (HSP) and also related to neurological diseases. However, the fact that crystallin serves as a disease marker in cervical cancer has not been found so far, and the present invention provides a method for detecting cervical cancer using the attenuation of crystallin expression as an index. As shown in the examples below, the expression of crystallin is specifically attenuated across individual differences in patients with cervical cancer, and by detecting this, cervical cancer is detected, or cervical cancer and other Can be distinguished from benign diseases in the uterus, such as benign tumors, uterine fibroids, and endometrial hyperplasia. The detection method may be appropriately selected and combined from those conventionally used, and is not intended to limit the present invention. However, labeling is performed using mRNA extracted from a test cell (or cDNA synthesized from mRNA) as a template. Labeling protein samples extracted from test cells, tissues, blood, body fluids, etc., or methods for detecting expression levels using probes consisting of complementary sequences, quantitative PCR using crystallin-specific primers A method of detecting cervical cancer by reacting an anti-crystallin antibody and qualitatively or quantitatively comparing with the expression of crystallin in a normal (= not cervical cancer) state obtained in advance is suitable.

  In the present specification, “attenuation of expression” of crystallin is biologically a state in which cancer tissue is down-regulated from a non-cancerous state, and as shown in the following Examples, protein expression in cervical cancer tissue and It is derived from the comparison with protein expression in non-cancerous tissue, but as a quantitative / numerical index, it is 30% or less of the normal state, suitably 25% or less, more suitably 20% or less. If so, it is appropriate to determine that the cancer is present. In comparison with the normal state, the expression in the patient's normal tissue may be compared with the expression in the tissue suspected of being cancerous. It may be digitized in a form such as, and comparison may be made between this and the expression level in a tissue suspected of having cancer. For quantitative comparison, the absolute amount of crystallin or the ratio per unit weight of protein may be obtained, but for simplification, immunodetection using anti-crystallin antibody is performed and the amount of antibody reacted (secondary antibody). It is better to substitute the light emission intensity of

  Further, as a possible embodiment of the present invention, there is a method for detecting cervical cancer using, as an index, increased expression of transgelin and / or protein disulfide isomerase (PDI) in addition to attenuation of crystallin expression. As shown in the examples below, transgelin and PDI are specifically expressed across individual differences in patients with cervical cancer. From test cells and tissues suspected of cancer, blood, body fluids, etc. By detecting the expression increase of transgelin and / or PDI in addition to the attenuation of crystallin expression in the extracted biological sample, more accurate cancer detection becomes possible. Regarding the expression of transgelin and PDI combined with the attenuation of crystallin expression, either one or both may be used. Similar to the above-described method for detecting the expression of crystallin, the expression detection method uses a nucleic acid or protein sample in a test cell or tissue, blood, body fluid, etc., and performs a qualitative or quantitative expression comparison with a non-cancerous state, A technique of detecting cervical cancer is suitable.

  The term “increased expression” of transgelin and / or PDI in the present specification is a state where biologically the cancer tissue is up-regulated from a non-cancerous state, and cervical cancer tissue as well as decreased expression of crystallin. It is derived from the comparison of protein expression in non-cancerous tissues and protein expression in non-cancerous tissues. As a quantitative and numerical indicator, in addition to the above-mentioned attenuation of crystallin expression, transgelin is 25% or more from the normal state. In addition to the above-mentioned decrease in the expression of crystallin in addition to an increase in the PDI, more suitably 30% or more, and / or an increase in the PDI of 15% or more from the normal state, more suitably 20% or more If there is an increase, it is desirable to determine that it is cancer. In comparison with the normal state, the expression in the patient's normal tissue may be compared with the expression in the tissue suspected of being cancerous. It may be digitized in a form such as, and comparison may be made between this and the expression level in a tissue suspected of having cancer. For quantitative comparison, the absolute amount of transgelin and / or PDI or the ratio per unit weight of protein may be obtained, but for simplification, immunodetection using anti-transgelin antibody and / or anti-PDI antibody It is preferable to substitute the amount of the reacted antibody (e.g., luminescence intensity of the secondary antibody).

  As a possible embodiment of the present invention, a suitable embodiment is a technique for detecting the expression of crystallin protein in a test cell. The method for detecting the expression of crystallin protein can be appropriately selected from the methods used as a standard method, but the detection method using antibodies is particularly suitable from the viewpoint of ease of quantitative comparison with non-cancerous cells. ing. Here, the anti-crystallin antibody is more suitably obtained by using the polypeptide described in SEQ ID NO: 1 (can be referred to in NCBI database: P02511) or a polypeptide having 95% or more homology with SEQ ID NO: 1 as an antigen. Antibodies, and more suitably labeled antibodies, which may be monoclonal or polyclonal as long as they have specificity for human crystallin protein. The production method may be any method as long as the polypeptide shown in SEQ ID NO: 1 or a partial sequence of the polypeptide is used. A method for detecting crystallin using these antibodies can be appropriately selected from conventional methods, and examples thereof include methods such as IF method and EIA method.

  As measures that can be taken by the present invention, in combination with the detection of crystallin expression using antibodies, anti-transgelin antibodies and / or anti-PDI antibodies are used to detect qualitatively or quantitatively the increase in the expression of these antigen proteins. Thus, it becomes possible to further improve the detection accuracy of cervical cancer using crystallin as a marker. Here, the anti-transgelin antibody is suitably obtained by using, as an antigen, the polypeptide described in SEQ ID NO: 2 (can be referred to in NCBI database: P37802) or a polypeptide having a homology of 95% or more with SEQ ID NO: 2. An antibody, or more suitably labeled antibody, may be a monoclonal or polyclonal antibody as long as it has specificity for human transgelin protein. Here, the anti-PDI antibody refers to an antibody obtained by using SEQ ID NO: 3 (referred to as NCBI database: P07237) or a polypeptide having 95% or more homology with SEQ ID NO: 3, and more appropriately labeled. The antibody may be a monoclonal antibody or a polyclonal antibody as long as it has specificity for human PDI protein. The method for detecting transgelin and / or PDI using these antibodies can also be appropriately selected from conventional methods, and examples thereof include methods such as IF method and EIA method.

  The present invention also provides a kit for detecting or diagnosing cervical cancer. The detection or diagnostic kit here includes, as a possible example, an anti-crystallin antibody (or anti-crystallin antibody-labeled secondary antibody complex) dissolved in an appropriate buffer, an antibody against a control protein, and staining. Each of which includes a reagent for detection, a blocking reagent, a solution for extracting a protein sample from cells / tissues, and a medium for mounting the extracted protein sample and reacting with an antibody. What is necessary is just to select suitably from the elements normally used about an element.

  The above detection or diagnostic kit may contain an anti-transgelin antibody and / or an anti-PDI antibody in addition to the anti-crystallin antibody. The anti-crystallin antibody herein is a suitable example of an antibody obtained by using the polypeptide of SEQ ID NO: 1 or a polypeptide having a homology of 95% or more with SEQ ID NO: 1 as an antigen. Is a suitable example of an antibody obtained using the polypeptide of SEQ ID NO: 2 or a polypeptide having a homology of 95% or more with SEQ ID NO: 2 as an antigen, and the anti-PDI antibody is a polypeptide of SEQ ID NO: 3. A suitable example is an antibody obtained using a peptide or a polypeptide having a homology of 95% or more with SEQ ID NO: 3 as an antigen.

  The present invention further provides a screening method for a cervical cancer therapeutic agent using increased expression of crystallin in a test cell as an index. As described above, in the cervical cancer cells, the expression of crystallin is attenuated to about 20% of the normal cells compared to the normal cells. This is contrary to the test cells such as cultured cervical cancer cells and cervical cancer cells. If a system is used to detect whether or not an increase in crystallin expression is observed in cells derived from precancerous lesion tissues collected from the cervical region, a specific and effective therapeutic agent for cervical cancer It will be possible to screen. In the screening process, the method for detecting crystallin expression may be appropriately selected from conventional methods, and the present invention is not limited thereto.

The present invention further provides a method for screening a therapeutic agent for cervical cancer using as an index the attenuation of transgelin and / or (PDI) expression in addition to the increased expression of crystallin in the test cells. As described above, expression of transgelin and PDI is increased by 20 to 30% in cervical cancer cells compared to normal cells, which is contrary to test cells such as cultured cervical cancer cells and uterus. In a cell derived from a precancerous lesion tissue collected from the cervix, a compound is allowed to act to detect whether or not transgelin and / or PDI expression is attenuated in addition to increased expression of crystallin, It will also enable screening for therapeutic agents that are more specific and effective for cervical cancer. In the screening process, the method for detecting transgelin and / or PDI expression may be appropriately selected from conventional methods, and the detection target may be both transgelin and PDI, or one of them.
Examples of the present invention are shown below, but the present invention is not limited to the examples.

(Comprehensive analysis of expressed proteins using clinical samples)
A total of 10 tissues, including 5 cases of normal cervical squamous epithelium (Group N) and 5 cases of squamous cell carcinoma (Group C) for which informed consent was obtained in writing for tissue collection and research use Each was homogenized, and the supernatant after centrifugation was used as a protein sample. As a specific procedure, the following procedure 1-4 was performed.
1. 1. Normal uterine vaginal squamous epithelium is collected from a specimen that has been hysterectomized for benign disease with no lesion in the uterine vagina, washed with physiological saline, and stored frozen at −80 ° C. Tissue was centrifuged with 1 ml solubilization solution (8M urea, 3% NP-40, appropriate amount of protease inhibitor in 40 mM Tris) on ice for 15 minutes under conditions of homogenization 3.150,000 rpm, 4 ° C. 3. Pass through a 0.45 μm filter and store at −80 ° C. Protein concentration is quantified using 2-D Quant Kit (GE healthcare)

The sample prepared as described above was separated by two-dimensional electrophoresis, and the spots were compared. Specifically, the procedure was as follows.
1st dimension IEF
A pH 3-10, 11 cm strip gel (Bio-rad) was used.
10 μl of sample (200 μg of protein amount) was added to the swelling buffer and swollen for 13.5 hours. Thereafter, electrophoresis was performed at 500 V, 1 h, 1000 V, 1 h, 8000 V, and 1 h 30 m.
Second dimension SDS-PAGE
After pretreatment with equilibration buffer and electrophoresis buffer, two polyacrylamide gels (12.5% acrylamide, 0.375M Tris, 0.1% SDS, 0.033% ammonium persulfate, 0.05% TEMED) were prepared. And migrated at a constant current of 30 mA.
Analysis CBB staining was performed and the expression of spots was observed.

  A common spot where the expression in group C was visually increased or attenuated as compared with group N was selected, and a spot where expression intensity was stable among individuals was selected. In addition, mass spectrometry performed peptide mass fingerprinting with MALDI-TOF-MS (using Agilent's LC-MS / MS with Spectrum Mill MS Proteomics Workbench), and protein identification was performed by Swiss-Prot database search. . Furthermore, the expression was confirmed by Western blotting using an antibody against the identified protein. Table 1 below summarizes the origin and characteristics of clinical specimens. In the table, N1-N5 and C1-C5 were subjected to two-dimensional electrophoresis, and the other samples were subjected to western blotting.

(Results) The results of protein expression comparison were as follows.
There was no individual difference in the pattern of proteins expressed in the N group. FIG. 1 shows a two-dimensional electrophoresis image (representative example) of a group N protein.
Compared with the N group, 27 spots were increased in expression in the C group, and 1 spot was attenuated. FIG. 2 shows a two-dimensional electrophoresis image (typical example) of the protein of group C.
As a result of analyzing a total of 7 spots (increase of 6 and decrease of 1) in which the expression intensity is stable among the spots having a difference between the N group and the C group, the expression of the 7 spots was found from the partial amino acid sequence information What was increased was a precursor of protein disulfide isomerase (PDI), heat shock protein β-1 (HSP27), transgelin, heat shock 70 kDa protein 1 (HSP70.1), keratin, type I cytoskeletal protein 19 (CK19), GAPDH, where the expression was attenuated was identified as alpha crystallin B chain (CRYAB; crystallin).

(Western blotting) Among the proteins identified in Example 1, focusing on crystallin (attenuation), transgelin, and PDI (increase) as proteins in which expression fluctuations in group C occur beyond individual differences. In order to confirm and quantify the expression state of the protein by Western blotting, the following experiment was conducted.
Each extract from the cells of Group N and Group C was electrophoresed on a 12.5% SDS-polyacrylamide gel with a protein amount of 100 μg. After the electrophoresis, it was transferred to a polyvinylidene fluoride (PVDF) film (manufactured by ATTO) by a semi-driving method. After blocking this membrane with a blocking solution (5% skim milk, 0.1% Tween-20, 1M Tris-HCl, pH 7.5), each antibody was used for antigen-antibody reaction,
The tissue extract is prepared by the same method as the two-dimensional electrophoresis method. 12.5% SDS-polyacrylamide gel electrophoresis was performed and transferred to a PVDF membrane (manufactured by ATTO). Antibodies against each protein as a primary antibody [anti-protein dissociated isomerase (PDI); (Santa Cruz Biotechnology), anti-transgelin-2; (ProteinTech), anti-αB-Crystallin (CRYAB) (CRYAB) ] And a secondary antibody, HRP (horseradish peroxidase) labeled antibody (DAKO) was used. The reaction was carried out by ECL Western blotting detection system (GE Healthcare), developed after exposure to film, and the expression intensity was compared.

The result of expression magnification (cancer / normal ratio) by densitometry was as follows.
PDI: 1.2 times or more Transgelin: 1.3 times or more CRYAB: less than 0.2 times That is, for PDI and transgelin identified as proteins whose expression is increased in Example 1, cervical cancer by densitometry CRYAB identified as a protein with increased expression 1.2 to 1.3 times normal in cells and attenuated expression is 0.2 times (1/5) normal in cervical cancer cells. It was shown that the expression was greatly attenuated until 1), confirming the results of the comprehensive analysis in Example 1.

(Immunostaining of cervical cancer with crystallin) Using the anti-crystallin antibody used in Example 2, immunostaining was performed using clinical tissues. The standard method was followed for the immunostaining protocol. FIG. 3 shows the results of immunostaining.
The regions indicated by arrows in the figure represent the stages of cancer, respectively, and from the left are dysplasia, carcinoma in situ, and microscopic invasive cancer, and microinvasive cancer is the most advanced stage. At the stage of dysplasia, the basal layer and middle upper layer of the cervical squamous epithelium showed staining (the part circled in the figure). On the other hand, these staining properties are remarkably reduced in the intraepithelial cancer region, and the process of disappearance of the staining property is clearly shown in the case of microinvasive cancer, indicating that the decrease in the expression of crystallin occurs in parallel with the progression of the cancer. It was done. The decrease in the staining property of crystallin is not only found to be different between normal and advanced cancer, but is considered to be effective in detecting very early cancers including carcinoma in situ.

  INDUSTRIAL APPLICABILITY The present invention can be used in the fields of cervical cancer, particularly squamous cell carcinoma markers and diagnostic kits, or screening for drug discovery search for cervical cancer therapeutics. In addition, as a method of using the present invention, in cases where the staining property of crystallin is reduced even at the stage of dysplasia, it is considered that the protein expression level has the character of cancer, and early treatment is recommended. This is thought to contribute to the determination of

Special table 2007-527001 Diagnostic markers for cancer Methods and compositions for detection of cervical cancer Marker for detecting uterine cancer Method for diagnosing endometriosis-related disease JP 2006-522607 Method and kit for diagnosing cervical cancer

1982 Annual Report of the Japan Obstetrics and Gynecology Association Gynecological Oncology Committee Report Report of the 2004 Annual Report of the Japan Obstetrics and Gynecology Association Gynecological Oncology Committee Gaarenstrom KN. et al. 2000. Gynecol. Oncol. 77 (1): 164-170. Pastner B.M. et al. 1989. Obset Gynecol. 74 (5): 786-788. Lehman M.M. 2001. Int. J. et al. Gynecol. Cancer 11 (2): 87-99. Gu Y. 2007. J. et al. Proteome Res. 6 (11): 4256-4268. Ono A.I. et al. 2008. Hum. Pathol. Aug. 26.

Claims (8)

A method for detecting cervical cancer, comprising detecting expression attenuation of an alpha crystallin B chain among crystallins in a biological sample. 2. The method according to claim 1, wherein the expression of alpha crystallin B chain in a biological sample is attenuated and the expression of transgelin and / or protein disulfide isomerase (hereinafter also referred to as PDI) is detected. The detection method of cervical cancer as described. The child according to claim 1 or 2, wherein an antibody obtained using the polypeptide of SEQ ID NO: 1 or a polypeptide having a homology of 95% or more with SEQ ID NO: 1 as an antigen in the detection of alpha crystallin B chain is used. A method for detecting cervical cancer. An antibody obtained by using the polypeptide shown in SEQ ID NO: 2 in detection of transgelin or a polypeptide having 95% or more homology with SEQ ID NO: 2 and / or the polypeptide shown in SEQ ID NO: 3 in detection of PDI Alternatively, the method for detecting cervical cancer according to claim 3, wherein an antibody obtained by using a polypeptide having 95% or more homology with SEQ ID NO: 3 as an antigen is used.   A kit for detecting or diagnosing cervical cancer for carrying out the method according to claim 1 or 2.   A kit for detecting or diagnosing cervical cancer, comprising the antibody according to claim 3 or 4. A screening method for a therapeutic drug for cervical cancer, using as an index the increase in expression of alpha crystallin B chain in a test cell. A screening method for a therapeutic drug for cervical cancer, using as an index an increase in alpha crystallin B chain expression in a test cell and a decrease in transgelin and / or protein disulfide isomerase (PDI) expression.