CN1952664A - Application of surface fatty acid-binding protein E-FABP - Google Patents

Abstract

The invention sifts the protein with differential expression which is in cancer organises of the hepatocyte cancer and nearby cancer organises to obtain a high expression protein in the cancer organizes of the hepatocyte cancer. The differential expression of the surface fatty acid binding protein E-FABP between the cancers organises of the hepatocyte cancer and nearby cancer organizes exists indeed is confirmed by the immune imprint experiment. In consideration of the correlation between the surface fatty acid binding protein E-FABP and the hepatocyte cancer, the protein can be used as theprotein molecule mark to detect the hepatocyte cancer.

Description

Application of Surface Fatty Acid Binding Protein E-FABP

technical field

The invention belongs to the field of biotechnology, in particular, the invention relates to the application of a surface fatty acid binding protein E-FABP as a protein molecular marker for detecting liver cancer.

Background technique

Liver cancer is a disease that seriously endangers human beings. The incidence of liver cancer in western developed countries is low, and the basic research on liver cancer in the world is still relatively weak. However, my country is a country with a high incidence of liver cancer, and the morbidity and mortality are on the rise, and the age of onset is younger. Medical expenses have increased greatly, and liver cancer has become the number one enemy that seriously endangers the safety of people's life and property in our country, and is an important factor affecting social and economic development. It is of strategic significance to intensify the basic research of liver cancer in my country, and to separate and identify new Liver cancer-related genes are a frontier topic in the basic research of liver cancer.

So far, the abnormal expression of no less than 20 genes has been determined to be related to the occurrence and development of liver cancer, but the abnormal expression rate of the identified liver cancer-related genes in liver cancer is not high, and the pathogenesis of liver cancer has not yet been elucidated. The early diagnosis rate still needs to be improved. In addition, traditional liver cancer surgery plus chemotherapy and multiple gene therapy methods used in combination in recent years have not significantly improved the survival rate of liver cancer patients. Therefore, finding new liver cancer-related genes, especially genes highly expressed in liver cancer, is of great importance for exploring the pathogenesis of liver cancer. significance.

Therefore, it is of great significance to study and develop genes and/or proteins highly expressed in liver cancer for therapeutic and diagnostic purposes. There is an urgent need in the art for new genes and/or proteins that are highly expressed in liver cancer.

Fatty acid-binding protein, epidermal; E-FABP; Psoriasis-associated fatty acid-binding protein homolog; PA-FABP; Fatty acid binding protein5; Psoriasis associated fatty acid binding protein homolog; Fatty acid binding protein 5, psoriasis associated; Fatty acid binding protein, psoriasis associated) Genebank accession number is gi|4557581, NCBI accession number is NP_001435, Swissprot accession number is Q01469, IPI number is: IPI00007797.1. Surface fatty acid-binding proteins belong to the family of fatty acid-binding proteins. Fatty acid-binding proteins (FABPs) are proteins with high affinity and specificity for long-chain fatty acids, and play a key role in the transport of long-chain fatty acids in cells. So far, at least three different fatty acid binding proteins have been discovered: B-FABP; E-FABP; H-FABP. The surface fatty acid binding protein we mentioned here is one of them. In addition to the functions mentioned above, this protein is considered to play a role in the differentiation process of keratinocytes (J.Neural.Transm.Suppl.2003;( 67): 225-34).

Fatty acid-binding protein (epidermal; E-FABP) was originally found to play an important role in preventing water permeability in the epidermis. Studies on mice with a phenotype of E-FABP deficiency found that, The histological level and growth of E-FABP-deficient mice are normal, while the expression of H-FABP in the liver is greatly increased (Mo1. Cell Biochem. 2002 Oct; 239(1-2): 83-6) . This suggests that H-FABP may have a complementary effect on E-FABP. Further studies on E-FABP-deficient homozygous mice showed that E-FABP does play a key role in the regulation of skin water permeability.

Recent work has carried out autoantibody antigen reaction research on the serum of patients with cancer (mainly brain cancer patients) and found that the high expression of E-FABP has been detected in the serum of more than 20% cancer patients, while in the serum of normal people, Only 2% of people have detected E-FABP, which indicates that E-FABP may be highly expressed in the serum of cancer patients (Biochem. Biophys. Res. Commun. 2004 Oct 8; 323(1): 156-62).

But so far, there is no report on the correlation between surface fatty acid binding protein E-FABP and hepatocellular carcinoma in the prior art.

Contents of the invention

By screening proteins differentially expressed in hepatocellular carcinoma tissues and paracancerous tissues of hepatocellular carcinoma, the inventors of the present application have found a protein that is differentially expressed in cancer tissues and paracancerous tissues of hepatocellular carcinoma (in cancer tissues) Up-regulated expression in tissues), which was identified as surface fatty acid binding protein E-FABP by mass spectrometry. Further immunoblotting experiments confirmed that the surface fatty acid binding protein E-FABP was indeed differentially expressed in the cancer tissues and adjacent tissues of HCC (upregulated expression in cancer tissues).

Based on the correlation between surface fatty acid binding protein E-FABP and hepatocellular carcinoma, using this protein as a protein molecular marker to detect its expression level can be used to detect liver cancer.

Therefore, the primary purpose of the present invention is to provide an application of a surface fatty acid binding protein E-FABP as a protein molecular marker for detecting liver cancer.

Another object of the present invention is to provide an anti-surface fatty acid binding protein E-FABP antibody, including monoclonal antibody and polyclonal antibody, for the preparation of a preparation for detecting liver cancer.

Another object of the present invention is to provide an anti-surface fatty acid binding protein E-FABP antibody, including monoclonal antibody and polyclonal antibody, for the preparation of a kit for detecting liver cancer.

Yet another object of the present invention is to provide a method for detecting whether the expression of the surface fatty acid binding protein E-FABP in the liver cell tissue is abnormal in vitro, the method comprising the following steps:

A, detect the quantity of surface fatty acid binding protein E-FABP in the hepatocyte to be tested with the antibody of specificity anti-surface fatty acid binding protein E-FABP;

B. Compare the amount of surface fatty acid binding protein E-FABP measured in step A with the amount of surface fatty acid binding protein E-FABP in normal liver tissue, if the amount of protein measured is higher than the normal value, it means that it has been detected Abnormal expression of surface fatty acid binding protein E-FABP in liver tissue.

Although in the prior art there are reports related to surface fatty acid binding protein E-FABP and cancer, so far, there is no report on the correlation between surface fatty acid binding protein E-FABP and hepatocellular carcinoma. Therefore, the present invention This discovery will provide a new avenue for the diagnosis and/or treatment of HCC.

Description of drawings

Figure 1 shows the results of western blot analysis of the surface fatty acid binding protein E-FABP, Beta-actin as a control.

Figure 2 shows the results of immunoblotting analysis of surface fatty acid binding protein E-FABP in serum samples of 36 randomly selected patients with liver cancer, serum samples of 24 patients with hepatitis B but not liver cancer, and 36 normal human serum samples, transferrin was used as a control; Among them, N stands for normal human serum, I stands for serum of patients with hepatitis B but not liver cancer, and C stands for serum of patients with liver cancer.

Fig. 3 is a relative distribution diagram of protein expression obtained by data analysis of the western blot pattern in Fig. 2 using R data analysis software.

Detailed ways

Below in conjunction with specific embodiment, further illustrate the present invention. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention.

The experimental methods not indicating specific conditions in the following examples are usually according to conventional conditions such as Sambrook et al., molecular cloning: the conditions described in the laboratory manual (New York: Cold Spring Harbor Laboratory Press, 1989), or according to the manufacturer's suggested conditions.

The inventors used nonenzymatic sample preparation (nonenzymatic sample preparation, NESP) to prepare the protein samples of hepatocellular carcinoma tissue and paracancerous tissue, and used isotope affinity labeling and gel-enhanced liquid chromatography-mass spectrometry (gel-enhanced Liquid chromatography-mass spectrometry coupled with isotope-coded affinitytag (GeLC-MS-ICAT) identified the proteins and compared their expression levels, and found that the surface fatty acid binding protein E-FABP was highly expressed in HCC tissues. Western blotting further confirmed that the surface fatty acid binding protein E-FABP was differentially expressed in HCC tissues and adjacent tissues.

Therefore, detecting the expression level of the surface fatty acid binding protein E-FABP as a protein molecular marker can be used to detect liver cancer, that is, the surface fatty acid binding protein E-FABP can be used as a protein molecular marker for detecting liver cancer.

Example 1. Preparation of Protein Samples of Hepatocellular Carcinoma Tissue and Paracancerous Tissue

Urea, 3-[(3-cholamidopropyl)-diethylammonium]-1-propanesulfonic acid (CHAPS), phenylmethylsulfonyl fluoride (PMSF), dithiothreitol used in this example (DTT) were purchased from Sigma Company.

In this embodiment, protein samples of cancer tissue and paracancerous tissue of hepatocellular carcinoma were prepared by nonenzymatic sample preparation (NESP), as follows:

The surgically excised fresh tissue block is quickly placed on ice and quickly cut into several small pieces that are visible to the naked eye without areas of necrosis. Use pre-cooled glutamine-free RPMI1640 medium (5% fetal bovine serum, 0.2mM PMSF, 1mM EDTA, benzisoxazole penicillin 25mg/mL, gentamicin 50mg/mL, penicillin 100U/mL, Streptomycin 100mg/mL, Amphotericin B 0.25mg/mL, Nystatin 50U/mL) After washing the small tissue pieces several times, they were quickly ground into cell pellets in liquid nitrogen, and the cell pellets were dissolved in an appropriate amount of lysate ( 8mol/L urea, 4% CHAPS, 40mmol/L Tris, and 65mmmol/L DTT), ultrasonic cell disruptor (Soniprep 150, UK, MSE) was used for intermittent ultrasonication in ice bath for 2min, centrifuged at 15000r/min, 4°C for 1h. The supernatant was taken, and the total protein was quantified by the modified Bradford method (see the product manual of Bio-Rad Company). The prepared protein samples of the hepatocellular carcinoma tissue and the corresponding paracancerous tissue were aliquoted and stored at -80°C for future use.

Eleven pairs of protein samples of hepatocellular carcinoma tissues and paracancerous tissues were prepared by the above method. The 11 cases of hepatocellular carcinoma specimens were all obtained from Dongfang Hepatobiliary Surgery Hospital, and were confirmed as hepatocellular carcinoma by 2 pathologists. All were male, with an average age of 48.5 years (31-65 years old). The serum test was positive for hepatitis B virus infection, and 11 cases (100%) belonged to clinical grade (TNM grade) III. Among them, 10 cases (90.9%) had alpha-fetoprotein (AFP) higher than 25 μg/L; 9 cases had tumors larger than 5 cm. The pathological data of 11 cases of HCC specimens are shown in Table 1.

Table 1. Pathological data of 11 cases of hepatocellular carcinoma specimens

No. gender age HBV HCV level AFP size 327 male 44 + - III ＞1000 8×8×7 415 male 40 + - III ＞1000 10×8×6 418 male 31 + - III 3.7 8×5×8 422 male 57 + - III ＞1000 3.5×4 429 male 44 + - III ＞1000 7.2×6 317 male 58 + - III ＞1000 5.2×6.4 42 male 45 + - III ＞1000 7.7×5.4 45 male 51 + - III ＞1000 5.5×4.0 48 male 55 + - III ＞1000 4×3 49 male 43 + - III ＞1000 12×12 424 male 65 + - III ＞1000 11.5×6.5

The cancer tissue and paracancerous tissue samples used in this embodiment are paired samples taken from the same hepatocellular carcinoma patient, and all 11 routine hepatocellular carcinoma cases have very similar case diagnostic indicators: they are all males, with an average age of 48.5 years (31 years old). ~65 years old), the serum test was positive for hepatitis B virus infection, and 11 cases (100%) belonged to TNM grade III. Among them, 10 cases (93.75%) had AFP higher than 25 μg/L; 9 cases had tumors larger than 5 cm. This sampling method is conducive to reducing the impact of individual differences on experimental analysis.

Example 2 , Screening of Differentially Expressed Proteins

Urea, 3-[(3-cholamidopropyl)-diethylammonium]-1-propanesulfonic acid (CHAPS), sodium dodecylsulfonate (SDS), dithiothreitol used in this example (DTT) was purchased from Sigma; iodoacetamide (IAA), acrylamide, N, N-methylenebisacrylamide, etc. were purchased from Fluka; cleavable ICAT reagents were purchased from Applied Biosystems Framingham, MA.

Ammonium persulfate (AP), TEMED, Tri-n-butylphosphat (TBP), PDQuest software, etc. are Bio-Rad products.

Avidin affinity columns were purchased from Applied Biosystems, Framingham, MA.

LCQ ^™ Deca XP system and ProteomeX ^™ Workstation were purchased from Thermo Finnigan.

The composition of the loading buffer used is: 0.6ml of 1mol/L Tris-HCl (pH6.8), 5ml of 50% glycerol, 2ml of 10% SDS, 0.5ml of mercaptoethanol, 1.9ml of distilled water, and a small amount of hexabromophenol blue.

First, the gel-enhanced liquid chromatography-mass spectrometry coupled with isotope-coded affinity tag (GeLC-MS-ICAT) was used to analyze the 11 pairs of protein samples obtained in Example 1. One pair (case number 429) of the differentially expressed proteins in the cancer tissue and paracancerous tissue of hepatocellular carcinoma was screened and identified, and the method was referred to the literature published on MCP by Steven P. Gygi in 2003 (Jiaxu Li et al.Mol Cell Proteomics.2003 Nov; 2(11):1198-204.Epub.2003 Sep.23), the specific process is as follows:

A pair of hepatocellular carcinoma tissue and corresponding paracancerous tissue protein samples prepared by the NESP method (case number 429), respectively took 100 μg, first reduced the protein with TBP, and then labeled with cleavable ICAT reagents (C ₁₂ and C ₁₃ ) respectively ( Among them, C ₁₂ and C ₁₃ correspond to cancer tissue and corresponding paracancerous tissue respectively, and the labeling method refers to the product manual). After mixing, add an appropriate amount of loading buffer, run 5% stacking gel (upper gel) and 7.5% ~ 17.5% separating gel (lower gel), electrophoresis conditions are 15mA/gel for 30min, then 30mA/gel, keep until the front of bromophenol blue Put about 8cm into the stacking gel.

After dyeing the strips, cut the entire loading strip into 8 parts on average, and cut each part into small pieces of 1mm ³ , decolorize in 100mM NH ₄ HCO ₃ , 30% ACN, vacuum freeze-dry, 100μl Place in 50mmol/L NH ₄ HCO ₃ (pH8.3, protein:trypsin=1:5, w/w) at 4°C for 2 hours, add 50μl 50mmol/L NH ₄ HCO ₃ (pH8.3), and digest overnight at 37°C .

Protein was extracted (60% ACN, 0.1% TFA), and vacuum freeze-dried. Each peptide mixture after enzymatic hydrolysis was purified by Avidin affinity column to obtain labeled peptides, and then used LCQ ^TM ProteomeX ^TM Workstation for liquid phase tandem (LC-MS/MS) mass spectrometry identification, Bioworks (Thermo finnigan company) software Database searches were performed and numerical calculations were performed with relex software (Scripps Research Institute, USA).

Using NESP method and GeLC-MS-ICAT technology, we identified 426 proteins with quantitative relationship. Among them, 201 proteins were quantitatively changed more than twice, and 155 proteins were highly expressed in HCC tissues; 46 proteins were highly expressed in hepatocellular carcinoma paracancerous tissues.

Using LC-MS/MS mass spectrometry identification, database search and ratio calculation, two Cys-containing peptides were identified for a total of 3 times, and the surface fatty acid-binding protein (E-FABP; Psoriasis-associated Fatty acid-binding protein homolog; PA-FABP; Fatty acid binding protein5; Psoriasis associated fatty acid binding protein homolog; Fatty acid binding protein 5, psoriasis associated; Fatty acid binding protein, psoriasis associated) were consistent, and the amino acid coverage was 15.56%. The calculation results of relex software showed that the surface fatty acid binding protein E-FABP was highly expressed in hepatocellular carcinoma tissues, and the ratio of cancer tissue/paracancerous tissue was 2.682 (SD=0). The detailed identification and peptide scoring results are shown in Table 2( C ^* in the table represents C ₁₃ labeled Cys).

Table 2. Detailed identification results of two Cys-containing peptides in GeLC-MS-ICAT

 Identified peptides (9 times in 4 segments) Mass number (MH+) charge number X corr value Delta Cn value K.TTQFSC*TLGEK.F   1451.62 2 2.237 0.3217 K.TTQFSC*TLGEKFEETTADGRK.T   2586.81 3 3.903 0.4343 K.TTQFSCTLGEK.F 1442.62 2 2.3031 0.1362

Example 3. Western blot verification of differential expression of surface fatty acid binding protein E-FABP

In order to confirm the differential expression of surface fatty acid binding protein E-FABP, 10 hepatocellular carcinoma patients' cancer tissues and corresponding paracancerous tissue protein samples (prepared by NESP method, other 10 cases except 429 cases in Table 1) were taken, purchased with The anti-surface fatty acid binding protein E-FABP antibody was used for western blot analysis, and the specific process was briefly described as follows:

Take 60 μg of protein samples for each sample and separate them with sodium dodecylsulfonate-polyacrylamide gel (i.e. SDS-PAGE, gel concentration 12%), transfer to PVDF membrane (purchased from Amersham Biosciences), primary antibody Use rabbit anti-human surface fatty acid binding protein E-FABP monoclonal antibody (purchased from HyCult biotechnology b.v. company, 1:100 dilution), incubate at room temperature for 2 hours, and use TBST (containing 2.42g of Tris per liter, 8g of sodium chloride, 20ml of Tween, Adjust the pH to 7.6 with HCl) and wash three times, 5 minutes each time, and incubate at room temperature for 1 hour as the secondary antibody (purchased from Santa Cruz Company, diluted 1:5000), then wash three times with TBST, 10 minutes each time, and finally After reacting with ECL plus reagent (Amersham Biosciences) for 5 minutes, it was detected by X-ray film exposure; at the same time, beta-actin was used as the control of the same amount of loading (beta-actin monoclonal antibody was purchased from abcam company, diluted 1:1000, the secondary antibody for mice), and the results are shown in Figure 1.

The results of Western blot in Figure 1 show that the 10 pairs of cancer tissues and para-cancerous tissues are basically loaded with the same amount, and this phenomenon is present in the 10 pairs of cancer tissues and para-cancerous tissues: the surface fatty acid binding protein E-FABP in cancer tissues The concentrations of the hybridization bands were significantly higher than those in the corresponding paracancerous tissues; it can be seen that the surface fatty acid binding protein E-FABP was highly expressed in the cancer tissues of hepatocellular carcinoma, which was consistent with the results of mass spectrometry.

In order to reflect better application value, we randomly selected the serum of 36 patients with liver cancer (see Table 3 for sample information), the serum of 24 patients with hepatitis B but not liver cancer (see Table 4 for sample information), and the serum of 36 normal people. (see Table 5 for sample information), and randomly grouped (each group includes one case of normal human serum, one case of serum from a patient with hepatitis B but not liver cancer, and one case of serum from a patient with liver cancer), 10ug protein samples were taken from each sample, and the above-mentioned immunoblotting method was also used The detection was carried out, and transferrin was used as the control of the same amount of loading (transferrin monoclonal antibody was purchased from abcam company, diluted 1:1000, and the secondary antibody was mouse). The results are shown in Figure 2.

At the same time, we used the R data analysis software (downloaded from http://www.r-project.org/ for free) to analyze the data of the immunoblotting pattern in Figure 2, and obtained the relative distribution of protein expression, the results are shown in Figure 3 shown.

It can be seen from Figure 2 and Figure 3 that the loading amount of all samples is basically the same (this conclusion can be drawn from the transferrin in Figure 3), but the expression of E-FABP protein in the serum of liver cancer patients is significantly higher than normal The amount in human serum shows that the surface fatty acid binding protein E-FABP is highly expressed in the serum of patients with hepatocellular carcinoma, which is consistent with the results of mass spectrometry.

In summary, the surface fatty acid binding protein E-FABP is differentially expressed in the cancer tissues and paracancerous tissues of hepatocellular carcinoma, which is obviously closely related to the occurrence and development of hepatocellular carcinoma. Therefore, the surface fatty acid binding protein E-FABP - FABP is used as a protein molecular marker to detect its expression level and can be used to detect hepatocellular carcinoma. Correspondingly, antibodies specific to the surface fatty acid binding protein E-FABP include various monoclonal antibodies and polyclonal antibodies against the surface fatty acid binding protein E-FABP, because they can be used to detect the surface fatty acid binding protein E-FABP Therefore, it can be used to detect liver cancer, or to prepare preparations or kits for detecting liver cancer, which is obvious to those skilled in the art.

Although the biological function and tumor-related mechanism of surface fatty acid binding protein E-FABP dynamics need to be further studied, it is certain to use it as a marker for detecting liver cancer. Surface fatty acid binding protein E-FABP can be used as a potential marker of hepatocellular carcinoma, and its biological function in cells suggests that surface fatty acid binding protein E-FABP may be used as a molecular marker for prognosis of liver cancer and a target molecule for clinical treatment.

Table 3. Data of liver cancer serum samples

No. gender age HBV grade AFP(ug/L) C1 male 32 + III Unknown C2 female 51 + IV Unknown C3 Unknown Unknown Unknown Unknown Unknown C4 male 34 + Unknown Unknown C5 female 50 - Unknown Unknown C6 female 63 + Unknown Unknown C7 male 76 + Unknown Unknown C8 male 70 Unknown Unknown C9 male 47 + Unknown Unknown C10 male 55 - Unknown Unknown C11 male 76 + Unknown Unknown C12 male 63 + Unknown Unknown C13 male 52 + III Unknown C14 female 44 Unknown III Unknown C15 Unknown Unknown Unknown Unknown Unknown C16 female 42 - Unknown 4.8 C17 male 34 + III 58.9 C18 female 71 - Unknown 3.3 C19 male 61 + Unknown Unknown C20 female 49 Unknown Unknown Unknown C21 female 76 - Unknown 22.3 C22 male 73 - Unknown 17.7 C23 male 81 + Unknown - C24 male 61 + Unknown 1000 C25 male 68 - Unknown 1.3 C26 male 67 + Unknown Unknown C27 male 49 + Unknown 146.5 C28 male 55 + III 837 C29 male 50 + II 27.3 C30 male 29 + III 1000 C31 male 42 + III Unknown C32 male 68 + III Unknown C33 male 69 + Unknown - C34 male 29 + Unknown 1000 C35 male 42 + Unknown 297.6 C36 female 65 + Unknown 59.3ug/L

Table 4. Hepatitis B serum sample information

No. gender age HBV I1 male 39 + I2 male 35 + I3 male 46 + I4 male 54 + I5 male twenty four + I6 male 41 + I7 male twenty two + I8 male 46 + I9 male 33 + I10 male 45 + I11 male 37 + I12 male 48 + I13 male 34 + I14 male 40 + I15 male 44 + I16 male 13 + I17 male 44 + I18 male 45 + I19 male 28 + I20 male 30 + I21 female 33 + I22 male 38 + I23 male 55 + I24 male 29 +

Table 5. Data of normal serum samples

No. gender age N1 male twenty two N2 male 25 N3 male 26 N4 male 26 N5 male 25 N6 male 27 N7 male 28 N8 male twenty four N9 male 29 N10 female 20 N11 female 26 N12 female 25 N13 female twenty three N14 female 26 N15 female twenty four N16 female twenty two N17 female 25 N18 female twenty three N19 female 25 N20 female 31 N21 female 26 N22 female twenty three N23 female 27 N24 female twenty three N25 female twenty two N26 female 28 N27 female twenty two N28 female twenty three N29 female twenty three N30 female twenty four N31 female twenty two N32 female twenty three N33 female 25 N34 female 25 N35 female 26 N36 female 28

Claims (9)

1. The application of a surface fatty acid binding protein E-FABP, characterized in that it is used as a protein molecular marker for detecting liver cancer. 2. The application according to claim 1, characterized in that the protein molecular marker used to detect liver cancer is to detect the expression level of the protein in liver cell tissue. 3. The application according to claim 2, wherein said detecting the expression level of the protein in the liver cell tissue is detecting whether the protein is up-regulated in the liver cell tissue. 4. The use of an antibody against surface fatty acid binding protein E-FABP, characterized in that it is used for preparing a preparation for detecting liver cancer. 5. The use according to claim 4, wherein the antibody against surface fatty acid binding protein E-FABP includes monoclonal antibody and polyclonal antibody. 6. The use of an antibody against surface fatty acid binding protein E-FABP, characterized in that it is used for preparing a kit for detecting liver cancer. 7. The use according to claim 6, wherein the antibody against surface fatty acid binding protein E-FABP includes monoclonal antibody and polyclonal antibody. 8. A method for detecting in vitro whether the expression of the surface fatty acid binding protein E-FABP in liver cell tissue is abnormal, characterized in that it comprises the following steps: A, detect the quantity of surface fatty acid binding protein E-FABP in the hepatocyte to be tested with the antibody of specificity anti-surface fatty acid binding protein E-FABP; B. Compare the amount of surface fatty acid binding protein E-FABP measured in step A with the amount of surface fatty acid binding protein E-FABP in normal liver tissue, if the amount of protein measured is higher than the normal value, it means that it has been detected Abnormal expression of surface fatty acid binding protein E-FABP in liver tissue. 9. The method according to claim 8, wherein said antibody against surface fatty acid binding protein E-FABP comprises monoclonal antibody and polyclonal antibody.

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