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

The application of surface fatty acid-binding protein E-FABP

Technical field

The invention belongs to biological technical field, specifically, the present invention relates to of the application of a kind of surface fatty acid-binding protein E-FABP as the protein molecular marker that detects liver cancer.

Background technology

Liver cancer is a kind of serious harm human diseases.The incidence of disease of western developed country liver cancer is lower, still comparatively weak to the fundamental research of liver cancer in the world, and China country occurred frequently that is liver cancer, M ﹠ M presents ascendant trend, and age of onset constitutes rejuvenation, the medical expense that is used for liver cancer treatment every year greatly increases, liver cancer has become serious harm China people life property safety's dead enemy, and be a key factor that influences socio-economic development, the fundamental research of going into overdrive to carry out China's liver cancer has strategic importance, and separates and identify that new liver cancer related gene is the advanced subject in the present liver cancer fundamental research.

Up to the present, the gene unconventionality expression that does not have 20 kinds is determined relevant with the generation development of liver cancer, but the unconventionality expression rate of fixed liver cancer related gene in liver cancer is not high, and the pathogenesis of liver cancer is not illustrated so far yet, and the early diagnostic rate of liver cancer still remains to be improved.In addition, traditional operation of liver cancer adds chemotherapy and the several genes methods of treatment that is used does not in recent years still have obviously to improve the survival rate of liver cancer patient, thereby especially liver cance high-expression gene is significant for the pathogenesis of inquiring into liver cancer to seek new liver cancer related gene.

Therefore, to research and develop in liver cancer the gene and/or the albumen of high expressed significant for treatment and diagnostic purpose.This area press for new in liver cancer the gene and/or the albumen of high expressed.

Surface fatty acid-binding protein (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, psoriasisassociated; Fatty acid binding protein, psoriasis associated) Genebank accession number is gi|4557581, and the accession number of NCBI is NP_001435, and the Swissprot accession number is Q01469, is for IPI number: IPI00007797.1.Surface fatty acid-binding protein belongs to fatty acid binding protein family, and fatty acid binding protein (FABPs) is a kind of and long-chain fatty acid has highly affine specific albumen, and the transhipment to long-chain fatty acid plays critical effect in cell.Up to the present, three kinds of different fatty acid binding proteins: B-FABP have been found at least; E-FABP; H-FABP.Here the surface fatty acid-binding protein that mentioned is exactly wherein a kind of, and except above said function, this albumen is considered to bringing into play effect (J.Neural.Transm.Suppl.2003 in the atomization of horn cell; (67): 225-34).

Surface fatty acid-binding protein (Fatty acid-binding protein, epidermal; E-FABP) be found at first in the epidermis and stop the permeability of moisture to play an important role, to phenotype is that the mouse of E-FABP defective is discovered, the histological level of E-FABP deficient mice and growth all are normal, and the expression of H-FABP (the Mo1.Cell Biochem.2002 Oct that raises greatly in the liver; 239 (1-2): 83-6).This shows that H-FABP may have additional effect to E-FABP.The homozygote mouse of E-FABP deficiency be studies show that further E-FABP plays critical effect at skin in to the adjusting of moisture permeability really.

Having work that cancer (mainly being cancer of the brain patient) patient's serum is carried out the autoantibody antigen-reactive recently discovers, detected the high expressed of E-FABP in the serum of cancer patient more than 20%, and in the serum to the normal person, have only 2% manual inspection to measure E-FABP, these data show E-FABP may be in cancer patient's serum high expressed (Biochem.Biophys.Res.Commun.2004 Oct 8; 323 (1): 156-62).

But up to the present, do not see report in the prior art relevant for the correlativity of surface fatty acid-binding protein E-FABP and hepatocellular carcinoma.

Summary of the invention

Protein by screening differential expression in hepatocellular carcinoma cancerous tissue and hepatocellular carcinoma cancer beside organism, the present inventor found a kind of in hepatocellular carcinoma cancerous tissue and cancer beside organism in there are differences expressed protein (up-regulated expression in cancerous tissue), be accredited as surface fatty acid-binding protein E-FABP through mass spectrum.Further immunoblot experiment confirms, surface fatty acid-binding protein E-FABP there are differences expression (up-regulated expression in cancerous tissue) really in the cancerous tissue of hepatocellular carcinoma and cancer beside organism.

Based on this correlativity of surface fatty acid-binding protein E-FABP and hepatocellular carcinoma, its expression is detected as a protein molecular marker with this albumen and can be used to detect liver cancer.

Therefore, primary and foremost purpose of the present invention promptly is to provide the application of a kind of surface fatty acid-binding protein E-FABP as the protein molecular marker that detects liver cancer.

Another object of the present invention is to provide a kind of antibody of anti-surface fatty acid-binding protein E-FABP, comprises monoclonal antibody and polyclonal antibody, is used to prepare the application of the preparation that detects liver cancer.

A further object of the present invention also is to provide a kind of antibody of anti-surface fatty acid-binding protein E-FABP, comprises monoclonal antibody and polyclonal antibody, is used to prepare the application of the kit that detects liver cancer.

Whether unusual another purpose of the present invention be to provide expression the method for surface fatty acid-binding protein E-FABP in a kind of vitro detection liver cell tissue, and this method may further comprise the steps:

A, with the quantity of surface fatty acid-binding protein E-FABP in the antibody test of the anti-surface fatty acid-binding protein E-FABP of the specificity liver cell to be measured;

The quantity of B, surface fatty acid-binding protein E-FABP that steps A is recorded and the quantity of the surface fatty acid-binding protein E-FABP in the normal liver tissue compare, as the albumen quantity that records is higher than normal value, then represents the abnormal expression of surface fatty acid-binding protein E-FABP in the detected hepatic tissue.

Though in the prior art relevant for the surface fatty acid-binding protein E-FABP report relevant with cancer, but up to the present, the report that does not also have the correlativity of surface fatty acid-binding protein E-FABP and hepatocellular carcinoma, therefore, this discovery of the present invention will provide a brand-brand-new way for the diagnosis and/or the treatment of hepatocellular carcinoma.

Description of drawings

Fig. 1 has shown the immunoblotting assay result to surface fatty acid-binding protein E-FABP, and Beta-actin is contrast.

Fig. 2 has shown to the 36 routine liver cancer patient blood serum samples of randomly drawing, 24 routine hepatitis B but not the immunoblotting assay result of surface fatty acid-binding protein E-FABP in the blood serum sample of liver cancer patient and the 36 routine normal human serum samples that transferrin is contrast; Wherein N represents normal human serum, and I represents hepatitis B but not liver cancer patient blood serum, and C represents liver cancer patient blood serum.

Fig. 3 carries out the relative distribution plan of protein expression amount that data analysis draws for adopting the R data analysis software to the Western blotting collection of illustrative plates of Fig. 2.

Embodiment

Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment only to be used to the present invention is described and be not used in and limit the scope of the invention.

The experimental technique of unreceipted actual conditions in the following example, usually according to people such as normal condition such as Sambrook, molecular cloning: laboratory manual (New York:Cold Spring Harbor Laboratory Press, 1989) condition described in, or the condition of advising according to manufacturer.

The inventor is with non-enzymolysis sample preparation method (nonenzymatic sample preparation, NESP) cancerous tissue of Zhi Bei hepatocellular carcinoma and cancer beside organism's protein example, LC-MS technology (gel-enhanced liquid chromatography-mass spectrometry coupled with isotope-coded affinitytag with isotope affinity tag and gel enhancing, GeLC-MS-ICAT) albumen is wherein identified and its expression relatively, be found that surface fatty acid-binding protein E-FABP high expressed in the hepatocellular carcinoma cancerous tissue.Immunoblot experiment confirms that further surface fatty acid-binding protein E-FABP there are differences expression really in the cancerous tissue of hepatocellular carcinoma and cancer beside organism.

Therefore, its expression being detected as a protein molecular marker with surface fatty acid-binding protein E-FABP and can be used to detect liver cancer, also is that surface fatty acid-binding protein E-FABP can be as the protein molecular marker that detects liver cancer.

Embodiment 1, hepatocellular carcinoma cancerous tissue and cancer beside organism's protein example preparation

Employed urea, 3-[(3-courage amido propyl in the present embodiment)-the diethyl ammonium]-1-propane sulfonic acid (CHAPS), phenylmethylsulfonyl fluoride (PMSF), dithiothreitol (DTT) (DTT) be all available from Sigma company.

Present embodiment with non-enzymolysis sample preparation method (nonenzymatic sample preparation, NESP) preparation hepatocellular carcinoma cancerous tissue and cancer beside organism's protein example, specific as follows:

The flesh tissue piece of excision places rapidly on ice, is cut into fast that several naked eyes are visible, the fritter of no necrotic zone.RPMI1640 nutrient culture media (5% hyclone that does not contain glutamine with precooling, 0.2mM PMSF, 1mM EDTA, oxacillin 25mg/mL, gentamicin 50mg/mL, penicillin 100U/mL, streptomysin 100mg/mL, amphotericin B 0.25mg/mL, nystatin 50U/mL) after washing organizes fritter for several times, in liquid nitrogen, grind to form cell precipitation fast, cell precipitation is dissolved in an amount of lysate (8mol/L urea respectively, 4%CHAPS, 40mmol/L Tris and 65mmmol/L DTT) in, (Soniprep 150, Britain for the ultrasonic cell disintegration instrument, MSE) ice bath ultrasonic 2min at intermittence, 15000r/min, 4 ℃ of centrifugal 1h.Get supernatant, it is quantitative to carry out gross protein with the Bradford method (seeing Bio-Rad company product description) of improvement, the hepatocellular carcinoma cancerous tissue for preparing and the protein example packing of corresponding adjacent tissues, and-80 ℃ of preservations are standby.

With 11 pairs of hepatocellular carcinoma cancerous tissues of method for preparing and cancer beside organism's protein example.11 routine hepatocellular carcinoma samples clearly are hepatocellular carcinoma all from east hospital of liver and gall surgical department by 2 doctors of pathology department.Be the male sex, 48.5 years old mean age (31～65 years old), serum detects the hepatitis B virus infections positive, and 11 examples (100%) belong to clinical scale (TNM classification) III level.Wherein, alpha-fetoprotein (AFP) is higher than 10 examples (90.9%) of 25 μ g/L; 9 routine tumours are greater than 5cm.The pathological data of 11 routine hepatocellular carcinoma samples sees table 1 for details.

The pathological data of table 1,11 routine hepatocellular carcinoma samples

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

The used cancerous tissue of present embodiment and cancer beside organism's sample are the paired samples of taking from same hepatocellular carcinoma patient, all 11 routine hepatocellular carcinoma cases have the case diagnosis index of fairly similar: be the male sex, 48.5 years old mean age (31～65 years old), serum detects the hepatitis B virus infections positive, and 11 examples (100%) belong to TNM classification III level.Wherein, AFP is higher than 10 examples (93.75%) of 25 μ g/L; 9 routine tumours are greater than 5cm.This sampling method helps reducing between individuality difference to the influence of experimental analysis work.

Embodiment 2, differentially expressed protein screening

The urea that uses in the present embodiment, 3-[(3-courage amido propyl)-the diethyl ammonium]-1-propane sulfonic acid (CHAPS), sodium dodecylsulphonate (SDS), dithiothreitol (DTT) (DTT) be available from Sigma company; Iodoacetamide (IAA), acrylamide, N, N-methylene diacrylamides etc. are available from Fluka company; Cleavable ICAT reagent is available from Applied BiosystemsFramingham, MA company.

Ammonium Persulfate 98.5 (AP), TEMED, Tri-n-butylphosphat (TBP), PDQuest software etc. are the Bio-Rad product.

The Avidin affinity column is available from Applied Biosystems, Framingham, MA company.

LCQ ^TMDeca XP system and ProteomeX ^TMWorkstation is available from Thermo Finnigan company.

Consisting of of employed sample-loading buffer: 1mol/L Tris-HCl (pH6.8) 0.6ml, 50% glycerine 5ml, 10%SDS 2ml, mercaptoethanol 0.5ml, distilled water 1.9ml, a small amount of six bromophenol blues.

LC-MS technology (the gel-enhanced liquidchromatography-mass spectrometry coupled with isotope-coded affinity tag that at first adopts isotope affinity tag and gel to strengthen, GeLC-MS-ICAT) cancerous tissue and the differentially expressed protein in the cancer beside organism of wherein a pair of (case the is numbered 4 29) hepatocellular carcinoma in 11 pairs of protein examples that embodiment 1 is obtained carry out Screening and Identification, and method is published in document (Jiaxu Li et al.Mol Cell Proteomics.2003 Nov on the MCP with reference to Steven P.Gygi in 2003; 2 (11): 1198-204.Epub.2003 Sep.23), detailed process is as follows:

A pair of hepatocellular carcinoma cancerous tissue and the corresponding adjacent tissues protein example (case is numbered 429) of NESP method preparation are got 100 μ g respectively, and be first with TBP crude protein also, then uses cleavable ICAT reagent (C respectively ₁₂And C ₁₃) mark (C wherein ₁₂With C ₁₃Corresponding cancerous tissue of difference and corresponding adjacent tissues, labeling method is with reference to product description).Mix the back and add an amount of sample-loading buffer, run 5% and concentrate glue (upper strata glue) and 7.5%～17.5% separation gel (lower floor's glue), deposition condition is 15mA/ glue 30min, and 30mA/ glue is retained to the bromophenol blue forward position and goes into to concentrate about glue 8cm then.

Examine dye band after, the whole batten band of going up on average is cut to 8 parts, every part is cut into 1mm more respectively ³Fritter, at 100mM NH ₄HCO ₃, decolour vacuum freeze drying, 100 μ l 50mmol/L NH among the 30%ACN ₄HCO ₃(pH8.3, protein: trypsase=1: 5, w/w) in 4 ℃ place 2hr, add 50 μ l 50mmol/LNH ₄HCO ₃(pH8.3), 37 ℃ of enzymolysis spend the night.

Extracting albumen (60%ACN, 0.1%TFA), vacuum freeze drying.Every part of peptide section potpourri behind the enzymolysis is used LCQ again after the Avidin affinity column is purified into mark peptide section ^TMProteomeX ^TMWorkstation carries out liquid phase series connection (LC-MS/MS) mass spectrum to be identified, Bioworks (Thermo finnigan company) software carries out database search and (Scripps Research Institute USA) carries out numerical evaluation with relex software.

Utilization NESP method and GeLC-MS-ICAT technology, we identify 426 kinds of protein that quantitative relationship is arranged altogether.Wherein the protein of the above quantitative change of twice is totally 201 kinds, hepatocellular carcinoma cancerous tissue high expressed 155 kinds of protein are arranged; Hepatocellular carcinoma cancer beside organism high expressed 46 kinds of protein are arranged.

Get 2 peptide sections that contain Cys with the evaluation of LC-MS/MS mass spectrum, database search and ratio calculation and identified altogether 3 times, with surface fatty acid-binding protein (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, psoriasisassociated; Fatty acid binding protein, psoriasis associated) conform to, the amino acid coverage rate is 15.56%.The relex computed in software is display surface fatty acid-binding protein E-FABP high expressed in the hepatocellular carcinoma cancerous tissue as a result, and the ratio of cancerous tissue/cancer beside organism is 2.682 (SD=0), and detailed evaluation situation and the marking of peptide section the results are shown in form 2 (C in the form ^*Represent C ₁₃The Cys of mark).

2 detailed qualification results that contain the peptide section of Cys among table 2, the GeLC-MS-ICAT

The peptide section that identifies (4 sections totally 9 times)	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

Embodiment 3, the surface fatty acid-binding protein E-FABP differential expression Western blotting checking

For confirming the differential expression of surface fatty acid-binding protein E-FABP, get 10 hepatocellular carcinoma patients' cancerous tissue and (the NESP method preparation of corresponding adjacent tissues protein example, other 10 example in the table 1 except that 429 examples), carry out immunoblotting assay with the anti-surface fatty acid-binding protein E-FABP antibody of buying, detailed process is summarized as follows:

Each sample is got 60 μ g protein examples with sodium dodecylsulphonate-polyacrylamide gel (that is: SDS-PAGE, gum concentration 12%) separates, be transferred on the pvdf membrane (available from Amersham Biosciences company), the one anti-anti-people's surface fatty acid-binding protein E-FABP of the rabbit monoclonal antibody that uses is (available from HyCult biotechnology b.v. company, dilution in 1: 100), incubated at room 2 hours, (every liter contains Tris 2.42g with TBST, sodium chloride 8g, Tween 20ml regulates pH to 7.6 with HCl) wash each 5 minutes three times, two anti-for anti-rabbit antibody (available from Santa Cruz company, dilution in 1: 5000) incubated at room is 1 hour, again with TBST washing three times, each 10 minutes, use ECL plus reagent (AmershamBiosciences) reaction after 5 minutes, at last with X-mating plate exposure tests; Simultaneously with beta-actin as the contrast of sample on the equivalent (the beta-actin monoclonal antibody is available from abcam company, dilution in 1: 1000, two anti-ly are mouse), the result is as shown in Figure 1.

The Western blotting result of Fig. 1 shows, the 10 pairs of cancerous tissues and cancer beside organism are sample on the equivalent basically, and all present such phenomenon in the 10 pairs of cancerous tissues and the cancer beside organism: the concentration of the hybridization band of surface fatty acid-binding protein E-FABP is all apparently higher than corresponding cancer beside organism in the cancerous tissue; There is high expressed in the visible surface fatty acid-binding protein E-FABP in the cancerous tissue of hepatocellular carcinoma, this result is consistent with the Mass Spectrometer Method result.

Be worth in order to embody better application, we at random picking the serum of 36 routine liver cancer patients (the sample data sees Table 3), 24 routine hepatitis B but not the serum (the sample data sees Table 4) of liver cancer patient, 36 routine normal human serums (the sample data sees Table 5), dividing into groups at random, (each group comprises a routine normal human serum, one routine hepatitis B but not liver cancer patient blood serum, one routine liver cancer patient blood serum), every routine sample is got the 10ug protein example, the above-mentioned western blotting method of same utilization detects, (the transferrin monoclonal antibody is available from abcam company as the contrast of sample on the equivalent to adopt transferrin simultaneously, dilution in 1: 1000, two anti-are mouse), the result is as shown in Figure 2.

Simultaneously, we adopt the R data analysis software (freely from Http:// www.r-project.org/Download) the Western blotting collection of illustrative plates to Fig. 2 carries out data analysis, draws the relative distribution plan of protein expression amount, and the result as shown in Figure 3.

As can be seen from Figures 2 and 3, the applied sample amount of all samples basic identical (from the transferrin of Fig. 3, can draw this conclusion), but the expression of E-FABP protein in liver cancer patient blood serum significantly is higher than the amount in the normal human serum, the visible surface fatty acid-binding protein E-FABP is high expressed in hepatocellular carcinoma patient's serum, and this result is consistent with the Mass Spectrometer Method result.

In sum, surface fatty acid-binding protein E-FABP there are differences expression in the cancerous tissue of hepatocellular carcinoma and cancer beside organism, generation development obvious and hepatocellular carcinoma has close correlativity, therefore, its expression is detected as a protein molecular marker with surface fatty acid-binding protein E-FABP and can be used to detect hepatocellular carcinoma.Accordingly, the antibody of the anti-surface fatty acid-binding protein E-FABP of specificity, the monoclonal antibody and the polyclonal antibody that comprise various anti-surface fatty acid-binding protein E-FABPs, because it can be used in the expression that detects surface fatty acid-binding protein E-FABP, thereby can be used to detect liver cancer, perhaps be used to prepare the preparation that detects liver cancer or kit etc., this is conspicuous for a person skilled in the art.

Though dynamic biological function of relevant surface fatty acid-binding protein E-FABP and tumour related mechanism are still waiting further research, but be sure as the label that detects liver cancer with it.Surface fatty acid-binding protein E-FABP can be used as the potential sign of hepatocellular carcinoma, and its biological function prompting surface fatty acid-binding protein E-FABP in born of the same parents may be as the prognosis molecule mark of liver cancer and the target molecule of clinical treatment.

Table 3, liver cancer serum sample data

No.	Sex	Age	HBV	Grade	AFP(ug/L)
						C1	The male sex	32	+	III	Not quite clear
C2	The women	51	+	IV	Not quite clear
						C3	Not quite clear	Not quite clear	Not quite clear	Not quite clear	Not quite clear
C4	The male sex	34	+	Not quite clear	Not quite clear
						C5	The women	50	-	Not quite clear	Not quite clear
C6	The women	63	+	Not quite clear	Not quite clear
						C7	The male sex	76	+	Not quite clear	Not quite clear
C8	The male sex	70		Not quite clear	Not quite clear
						C9	The male sex	47	+	Not quite clear	Not quite clear
C10	The male sex	55	-	Not quite clear	Not quite clear
						C11	The male sex	76	+	Not quite clear	Not quite clear
C12	The male sex	63	+	Not quite clear	Not quite clear
						C13	The male sex	52	+	III	Not quite clear
C14	The women	44	Not quite clear	III	Not quite clear
						C15	Not quite clear	Not quite clear	Not quite clear	Not quite clear	Not quite clear
C16	The women	42	-	Not quite clear	4.8
						C17	The male sex	34	+	III	58.9
C18	The women	71	-	Not quite clear	3.3
						C19	The male sex	61	+	Not quite clear	Not quite clear
C20	The women	49	Not quite clear	Not quite clear	Not quite clear
						C21	The women	76	-	Not quite clear	22.3
C22	The male sex	73	-	Not quite clear	17.7
						C23	The male sex	81	+	Not quite clear	-
C24	The male sex	61	+	Not quite clear	1000
						C25	The male sex	68	-	Not quite clear	1.3
C26	The male sex	67	+	Not quite clear	Not quite clear
						C27	The male sex	49	+	Not quite clear	146.5
C28	The male sex	55	+	III	837
						C29	The male sex	50	+	II	27.3
C30	The male sex	29	+	III	1000
						C31	The male sex	42	+	III	Not quite clear
C32	The male sex	68	+	III	Not quite clear
						C33	The male sex	69	+	Not quite clear	-
C34	The male sex	29	+	Not quite clear	1000
						C35	The male sex	42	+	Not quite clear	297.6
C36	The women	65	+	Not quite clear	59.3ug/L

Table 4, hepatitis B blood serum sample data

No.	Sex	Age	HBV
				I1	The male sex	39	+
I2	The male sex	35	+
				I3	The male sex	46	+
I4	The male sex	54	+
				I5	The male sex	24	+
I6	The male sex	41	+
				I7	The male sex	22	+
I8	The male sex	46	+
				I9	The male sex	33	+
I10	The male sex	45	+
				I11	The male sex	37	+
I12	The male sex	48	+
				I13	The male sex	34	+
I14	The male sex	40	+
				I15	The male sex	44	+
I16	The male sex	13	+
				I17	The male sex	44	+
I18	The male sex	45	+
				I19	The male sex	28	+
I20	The male sex	30	+
				I21	The women	33	+
I22	The male sex	38	+
				I23	The male sex	55	+
I24	The male sex	29	+

Table 5, normal serum sample data

No.	Sex	Age
			N1	The male sex	22
N2	The male sex	25
			N3	The male sex	26
N4	The male sex	26
			N5	The male sex	25
N6	The male sex	27
			N7	The male sex	28
N8	The male sex	24
			N9	The male sex	29
N10	The women	20
			N11	The women	26
N12	The women	25
			N13	The women	23
N14	The women	26
			N15	The women	24
N16	The women	22
			N17	The women	25
N18	The women	23
			N19	The women	25
N20	The women	31
			N21	The women	26
N22	The women	23
			N23	The women	27
N24	The women	23
			N25	The women	22
N26	The women	28
			N27	The women	22
N28	The women	23
			N29	The women	23
N30	The women	24
			N31	The women	22
N32	The women	23
			N33	The women	25
N34	The women	25
			N35	The women	26
N36	The women	28

Claims (9)

1, a kind of application of surface fatty acid-binding protein E-FABP is characterized in that, as the protein molecular marker that detects liver cancer.

2, application as claimed in claim 1 is characterized in that, described is to detect the expression of this albumen in the liver cell tissue as the protein molecular marker that detects liver cancer.

3, application as claimed in claim 2 is characterized in that, the expression of this albumen of described detection in the liver cell tissue is to detect this albumen whether to have up-regulated expression in the liver cell tissue.

4, a kind of application of antibody of anti-surface fatty acid-binding protein E-FABP is characterized in that, is used to prepare the preparation that detects liver cancer.

5, application as claimed in claim 4 is characterized in that, the antibody of described anti-surface fatty acid-binding protein E-FABP comprises monoclonal antibody and polyclonal antibody.

6, a kind of application of antibody of anti-surface fatty acid-binding protein E-FABP is characterized in that, is used to prepare the kit that detects liver cancer.

7, application as claimed in claim 6 is characterized in that, the antibody of described anti-surface fatty acid-binding protein E-FABP comprises monoclonal antibody and polyclonal antibody.

8, whether unusual the expression of surface fatty acid-binding protein E-FABP method in a kind of vitro detection liver cell tissue is characterized in that may further comprise the steps:

A, with the quantity of surface fatty acid-binding protein E-FABP in the antibody test of the anti-surface fatty acid-binding protein E-FABP of the specificity liver cell to be measured;

The quantity of B, surface fatty acid-binding protein E-FABP that steps A is recorded and the quantity of the surface fatty acid-binding protein E-FABP in the normal liver tissue compare, as the albumen quantity that records is higher than normal value, then represents the abnormal expression of surface fatty acid-binding protein E-FABP in the detected hepatic tissue.

9, method as claimed in claim 8 is characterized in that, the antibody of described anti-surface fatty acid-binding protein E-FABP comprises monoclonal antibody and polyclonal antibody.

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