CN1239700C - Human liver cancer related gene, encoded product and use thereof - Google Patents
Human liver cancer related gene, encoded product and use thereof Download PDFInfo
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- CN1239700C CN1239700C CN 02158110 CN02158110A CN1239700C CN 1239700 C CN1239700 C CN 1239700C CN 02158110 CN02158110 CN 02158110 CN 02158110 A CN02158110 A CN 02158110A CN 1239700 C CN1239700 C CN 1239700C
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Abstract
The present invention discloses a gene relevant to human liver cancer and an encoding product thereof. The gene is one of the following nucleotide sequences: 1) SEQ ID No. 1 in a coding sequence list; 2) a polynucleotide of the protein sequence as SEQ ID No. 2 or SEQ ID No. 3 in the coding sequence list; 3) a DNA sequence with more than 90 percent of homology with a DNA sequence limited by the SEQ ID No. 1 in the coding sequence list and for encoding a protein with the same function. The protein of the present invention can be used as a new index for the early diagnosis of the liver cancer; with the utilization of an ELISA method suitable for extensive clinical use and a prepared detection kit thereof, the early diagnosis rate of the primary liver cancer and the accuracy rate of the diagnosis can be improved; in addition, a gene LAPTM4B can be uses as a target gene of the treatment of the liver cancer, for example, the expression of the gene LAPTM4B can be depressed by siRNA interference technology as latest development, and the protein of LAPTM4B can be used as a novel target point of medicine actions for developing a new medicine with the target point of the protein of the LAPTM4B. Thereby, the possibility of developing a new approach for resisting the liver cancer exists, and the new approach for resisting the liver cancer is a project which can generating great social benefits.
Description
Technical field
The present invention relates to gene and coded product and application in the genetically engineered field, particularly relate to people's liver cancer related gene and coded product and application.
Background technology
(hepatocellular carcinoma is the major disease of serious harm people ' s health HCC) to hepatocellular carcinoma, it is reported, new patient's number in whole world primary hepatocarcinoma every year surpasses 1,000,000 people, wherein 70% concentrates on the Asia.China's liver cancer accounts for the 40-45% of whole world liver cancer, annual newborn liver cancer patient 450,000, and be continuous ascendant trend.Liver cancer is the sickness rate height not only, and concealment, progress are fast, recurrence rate and mortality ratio height, are called as " king in the cancer ".During the liver cancer patient of going to see a doctor mostly is greatly or late period, if do not treat natural history only 3-6 month.
The pathogenesis of illustrating liver cancer will help prevention, diagnosis and the treatment of liver cancer.Early diagnosis is the key that improves curative effect, reduces mortality ratio.At present used diagnosing cancer of liver mark AFP is negative at about 30% liver cancer patient, and the rising significantly of AFP can occur some optimum liver problem sufferers, causes the difficulty of differential diagnosis.
Tumour is the heredopathia of cell in itself.Though the gene relevant with liver cancer found that much liver cancer takes place, the mechanism of development is still fuzzy.The proto-oncogene of finding can be classified as four classes by its coded product substantially in the location and the function of cell at present, and a class is the gene of coding somatomedin, comprises sis, int-2, hst, fgf-5; Second class is the gene of coding growth factor receptors, comprises erbB, erbB-2, fms, met, ros etc.; The 3rd class is the gene of signal transduction molecule in the Codocyte matter, comprises abl, src, ras, raf, yes, fgr, fes, lck, mos etc.; The 4th class is the gene of Codocyte propagation and apoptosis regulation molecule, comprises bcl-1, bcl-2 etc.; The 5th class is the gene of the protein (transcription factor) that combines with DNA in the Codocyte nuclear, for example genes such as myc, myb, fos, jun, B-lym, ski, ets, rel.Studies confirm that and the closely-related ras of mainly containing of generation, src, myc, the genes such as met, p53 of HCC.
Summary of the invention
The purpose of this invention is to provide a new people's liver cancer related gene and a coded product thereof.
People's liver cancer related gene name provided by the present invention is called LAPTM4B, is one of following nucleotide sequences:
1) the SEQ ID № in the sequence table: 1;
2) SEQ ID № in the code sequence tabulation: 4 or SEQ ID №: the polynucleotide of 5 protein sequences.
SEQ ID № in the described sequence table: 1 by 954 based compositions, are complete reading frames, SEQ ID №: 1 has two initiation sites, and one is from 5 ' end 1-3 bit base, and another is from 5 ' end 274-276 bit base.Sequence SEQ ID №: 1 global cDNA has two, has different tailing signals, as SEQ ID №: 15 ' 85 bases that stretch out, 3 ' 401 bases that stretch out, promptly obtain the SEQ ID № in the sequence table: 2, this gene is by 1440 based compositions; As SEQ ID №: 15 ' 85 bases that stretch out, 3 ' 1130 bases that stretch out promptly obtain the SEQ ID № in the sequence table: 3, this gene is by 2169 based compositions.Gene LAPTM4B is positioned karyomit(e) 8q22.1.
Human hepatoma associated protein LAPTM4B, be to have sequence 4 in the sequence table or/and the protein of sequence 5 aminoacid sequences, or with sequence 4 or/and the aminoacid sequence of sequence 5 through replacement, disappearance or the interpolation of one or several amino-acid residue and have with sequence 4 or/and the aminoacid sequence of sequence 5 identical active by sequence 4 or/and sequence 5 deutero-protein.
SEQ ID №: 4 and SEQ ID №: 5 two kinds of protein all have four sections and wear the film sequence, and a possible N glycosylation site and typical lysosome signal for locating all belong to four times and wear the protein superfamily of film, but have the phosphorylation site that does not wait respectively.Experiment shows, SEQ ID №: 4 LAPTM4B may form mixture with growth factor receptors (as EGF-R) in the plasma membrane and integrin alpha 6 subunits (specific receptors of extracellular matrix ln), may have the proliferation signal that will come from somatomedin and extracellular matrix two aspects and be coupled at together function at plasma membrane.What this will further illustrate the growth of normal eukaryotic cell is deciding dependency (promptly except that the stimulus signal that need come from somatomedin, the stimulus signal that also needs to come from certain extracellular matrix could start cell proliferation) molecular mechanism, have breakthrough meaning on the regulatory mechanism of cell proliferation illustrating.Phosphorylation can take place in the tyrosine residues that experiment showed, LAPTM4B PROTEIN C end, thereby can form and intracellular signal molecule SH2 structural domain bonded site; Simultaneously, the N terminal sequence has the binding site of Pro enrichment region and typical SH3 structural domain.Indicate SEQ ID №: 4LAPTM4B albumen may be a docking protein that has vital role in signal transduction, can convene relevant signaling molecule to finish the signal transduction of cell proliferation.Experiment showed, the № with SEQ ID: 4 cDNA transfection mouse NIH 3T3cells, the propagation of transfectional cell is quickened as a result, serum dependence is reduced, and can generate the malignant fibrous sarcoma the NIH mouse.With SEQ ID №: 5 cDNA transfection human liver cancer cell HLE clone, transfectional cell are difficult to form the clone, and with SEQ ID №: the same clone of 4 cDNA transfection then can form big a large amount of clones.These results show SEQ ID №: 5LAPTM4B albumen and SEQ ID №: 4LAPTM4B albumen has the difference in functionality of neck mutually.
Gene LAPTM4B significantly raises and express in 96.88% (31/32) people's liver cancer tissue in the low expression of normal liver tissue, and visible expression is slight in the non-cancer hepatic tissue of portion paired raises.Expression level in most of hepatoma cell line is also higher.In addition, LAPTM4B also has in various degree wide expression in some tumor cell line and some healthy tissues of human body, and is wherein higher with the expression of testis, cardiac muscle and skeletal muscle.Contain SEQ ID №: 1, the expression vector of 2,3 described sequences and contain SEQ ID №: 1, the transfectional cell series of 2,3 sequences all belongs to protection scope of the present invention.
Protein of the present invention can be used as the New Set of hepatocarcinoma early diagnosis, is suitable for the ELISA method of widespread use clinically by employing, and prepared correlation detection test kit, can improve the early diagnostic rate and the accurate rate of diagnosis of primary hepatocarcinoma.Also the target gene of LAPTM4B gene as liver cancer treatment for example can be prevented the LAPTM4B expression of gene by the siRNA perturbation technique of recent development; Also can be with LAPTM4B albumen as pharmaceutically-active novel targets, development is the new drug of target spot with LAPTM4B albumen.Thereby in view of the above, might develop the new way of anti-liver cancer, this is an engineering that will produce great social benefit.
The present invention will be further described below in conjunction with specific embodiment and accompanying drawing.
Description of drawings
Fig. 1 analyzes collection of illustrative plates for Nortern Blot.
Fig. 2 is the slice map of immunocytochemical assay.
Fig. 3 analyzes collection of illustrative plates for Western Blot.
Fig. 4 analyzes collection of illustrative plates for Western Blot.
Fig. 5 is a cell growth curve.
Fig. 6 is a cell growth curve.
Fig. 7 is the oncogenic function of cDNA cells transfected of the present invention to mouse.
Fig. 8 is the LAPTM4B antigen levels histogram in the liver cancer patient blood serum.
Embodiment
Select four kinds of different hepatic tissues of proliferation/differentiation state, be normal adult liver (few propagation, end differentiation eventually), tire liver (vigorous propagation, low differentiation), liver cancer (propagation out of control, unusual differentiation) and match non-cancer hepatic tissue (liver cell before the cancer of active propagation) and carry out Nortern Blot and analyze, from 5 routine normal adult livers, 5 routine tire livers, 32 routine liver cancer, the fresh specimens of the non-cancer hepatic tissue of 32 example pairings are extracted RNA, through electrophoretic separation, be transferred to nylon membrane, hybridize with the probe of Dig mark, wash film, by specification demonstration hybridization signal in 68 ℃.The result as shown in Figure 1, wherein swimming lane 1 is the tire liver, swimming lane 2 is normal adult livers, swimming lane the 3,5,7, the 9th, liver cancer, swimming lane the 4,6,8, the 10th matches non-cancer hepatic tissue, the result shows that LAPTM4B is followed successively by at the expression level of various hepatic tissues: the non-cancer hepatic tissue of liver cancer＞pairing and tire liver＞normal adult liver.
The clone of embodiment 2, LAPTM4B
Select four kinds of different human hepatic tissues of proliferation/differentiation state, be normal adult liver (NL), tire liver (FL), liver cancer (HCC) and the non-cancer hepatic tissue (PNL) that matches, obtained the cDNA fragment (LC27) of unknown gene by the fluorescence differential display technique, with LC27 segment (426bp) serves as that the splicing of EST homologous sequence is carried out to 5 ' direction in the basis, and obtain the full length cDNA sequence of gene by RACE and high temperature RT-PCR experiment, be SEQ ID №: 2 and 3.
The location of embodiment 3, immunocytochemical assay LAPTM4B
Well-grown Bel7402 BEL-7402 cell is fixed on the slide glass room temperature 30 minutes with 4% Paraformaldehyde 96.PBS washes the back and acts on 15 minutes in order to eliminate endogenous peroxydase in containing 3% hydrogen peroxide in methanol.After 0.05%Tween 20/PBS rinsing, added 37 ℃ of 10% bovine serum albumin incubation sections 30 minutes.Overnight incubation in the specific antibody (polyclonal antibody of 10 peptides of anti-LAPTM4B, 1: 100) that adds suitable dilution, 4 ℃ of wet boxes add the goat anti-rabbit antibody of horseradish peroxidase-labeled after the PBS rinsing, hatched 30 minutes in 37 ℃ of wet boxes.The PBS rinsing with the DAB colour developing, is used Meyer ' s haematoxylin redyeing nuclear successively, conventional dehydration, transparent, sealing, and mirror is observed down.The result shows that LAPTM4B is positioned the membrane structure of cell as shown in Figure 2, comprises the cell inner membrance, may also have plasma membrane.
The Western Blot of embodiment 4, subcellular components analyzes
Respectively with HCC,, PNL and NL be organized in (150mM NaCl, 10mM Tris PH7.4,0.5%Triton X-100,5mM EDTA, 1mM PMSF) fully homogenate in the homogenate, through differential centrifugation separating nucleus, plastosome and membrane component.Three components are carried out the SDS-PAGE electrophoresis respectively, hybridize with the polyclonal antibody of LAPTM4B.The result as shown in Figure 3, wherein N is a nucleus, M is a cytolemma, Mit is a plastosome, swimming lane 1,2,3 is a liver cancer among the figure; Swimming lane 4,5,6 is the non-cancer hepatic tissue of pairing, and swimming lane 7,8,9 is the normal adult liver, and the result shows that LAPTM4B is present in the membrane component of cell, and is not present in nucleus and plastosome.
The Western Blot of embodiment 5, TritonX114 fractional extraction thing analyzes
With the people HCC of surgical resection, PNL and NL are organized in (150mM NaCl, 10mM TrisPH7.4,1%Triton X-114,5mM EDTA, 1mM PMSF) fully homogenate in the homogenate, centrifugal removal residue.Add 300 μ l and contain 6% sucrose in 200 μ l supernatant liquors, the TBS liquid of 0.06%Triton X-114 was hatched 5 minutes for 37 ℃.Be divided into water (upper strata) and de-sludging (lower floor) mutually after centrifugal, add 1%Triton X-114 at aqueous phase and act on 10 minutes on ice, hatched 5 minutes for 37 ℃, centrifugal back is collected water and is merged with a preceding water, and repetitive operation once.Add the TBS liquid washed twice that contains 2%Triton X-114 to water mutually respectively with de-sludging.The cold dehydrated alcohol precipitating proteins that adds two volumes then respectively is dissolved in sedimentary protein in the sample-loading buffer after centrifugal, carries out SDS-PAGE electrophoresis and Western blot and detects.The result as shown in Figure 4, swimming lane A is the homogenate of branch phase front among the figure, swimming lane B is the water after the phase-splitting.Swimming lane C is the de-sludging phase after the phase-splitting, and the result shows that LAPTM4B is present in the washing agent phase, illustrates that TAPTM4B albumen is present in the membranous structure of cell.
The regulating and controlling effect of embodiment 6, the gene pairs cell proliferation of the present invention of full-length cDNA transfection experiment proof
Structure contains the expression vector pcDNA-LAPTM4B of LAPTM4B full length gene cDNA sequence (2.16kb), transfection mouse NIH 3T3 cell, and the clone that the screening stability and high efficiency is expressed LAPTM4B measures cell count by the acid phosphatase enzyme process, draws cell growth curve; By the Flow cytometry cell cycle; Detect the expression level of cyclin with Western Blot.The result shows, with the NIH3T3 cell of the expression plasmid transfection that contains its full-length cDNA, cell proliferation is quickened, and as shown in Figure 5, shows that the expression of cyclin cyclinE obviously strengthens; Simultaneously, the propagation of transfectional cell reduces the dependency of serum (somatomedin), as shown in Figure 6, shows that this gene may participate in the regulation and control of cell proliferation, and it crosses expression (activation) may be out of control relevant with the propagation of cell.
Embodiment 7, cDNA cells transfected are to the oncogenic function of mouse
Get at random 6 age in week the NIH male mice be divided into three groups: the control group of physiological saline injection, the control group of the MOCK cell inoculation of unloaded plasmid transfection and contain the experimental group of the cell inoculation of full-length cDNA plasmid transfection.Every mouse is in right armpit subcutaneous vaccination 2 * 10
6Cell.Every group of 4-6 only.Disconnected neck is put to death after 21 days, dissects.The result can see that experimental group has two mouse to form the fibrosarcoma of tangible medium grade of malignancy (A, B) as shown in Figure 7, and two inoculation position is Lymphoid tissue (C, D) in addition.And 12 mouse of two control groups all do not have the tumour generation after inoculating 86 days.
The result and the express spectra thereof of embodiment 6 and example 7 show that LAPTM4B may be a new proto-oncogene.
Embodiment 8, with the LAPTM4B antigen levels in the ELISA initial analysis liver cancer patient blood serum
The every hole of 96 orifice plates is added the HCC patients serum or the normal human serum bag quilt of different extension rates, and 4 ℃ are spent the night.Each hole adds 2%BSA liquid and seals 1hr in room temperature with after containing the PBS liquid rinsing of 0.5%Tween-20.The anti-LAPTM4B10 peptide antibody of rabbit that adds different extension rates is in incubated at room 2hr, with adding the goat anti-rabbit antibody (1: 1000 times of dilution) of horseradish peroxidase-labeled in incubated at room 2hr after the rinsing of PBS liquid.Through the PBS rinsing, add the colour developing of 1 μ g/ml O-Phenylene Diamine after 10-15 minute, add H
2SO
4Termination reaction with microplate reader reading under the 490nm wavelength, detects antigen.The result shows as shown in Figure 8, has the LAPTM4B antigen high than normal people's level in the liver cancer patient blood serum.Indicate the New Set that LAPTM4B is expected to become diagnosing cancer of liver.
Sequence table
<160>5
<210>1
<211>954
<212>DNA
＜213〉Genus Homo people (Homo sapiens)
<400>1
atgacgtcac?ggactcgggt?cacatggccg?agtccgcccc?gccccctccc?cgtccccgcc 60
gctgcagccg?tcgccttcgg?agcgaagggt?accgacccgg?cagaagctcg?gagctctcgg 120
ggtatcgagg?aggcaggccc?gcgggcgcac?gggcgagcgg?gccgggagcc?ggagcggcgg 180
aggagccggc?agcagcggcg?cggcgggctc?caggcgaggc?ggtcgacgct?cctgaaaact 240
tgcgcgcgcg?ctcgcgccac?tgcgcccgga?gcgatgaaga?tggtcgcgcc?ctggacgcgg 300
ttctactcca?acagctgctg?cttgtgctgc?catgtccgca?ccggcaccat?cctgctcggc 360
gtctggtatc?tgatcatcaa?tgctgtggta?ctgttgattt?tattgagtgc?cctggctgat 420
ccggatcagt?ataacttttc?aagttctgaa?ctgggaggtg?actttgagtt?catggatgat 480
gccaacatgt?gcattgccat?tgcgatttct?cttctcatga?tcctgatatg?tgctatggct 540
acttacggag?cgtacaagca?acgcgcagcc?tggatcatcc?cattcttctg?ttaccagatc 600
tttgactttg?ccctgaacat?gttggttgca?atcactgtgc?ttatttatcc?aaactccatt 660
caggaataca?tacggcaact?gcctcctaat?tttccctaca?gagatgatgt?catgtcagtg 720
aatcctacct?gtttggtcct?tattattctt?ctgtttatta?gcattatctt?gacttttaag 780
ggttacttga?ttagctgtgt?ttggaactgc?taccgataca?tcaatggtag?gaactcctct 840
gatgtcctgg?tttatgttac?cagcaatgac?actacggtgc?tgctaccccc?gtatgatgat 900
gccactgtga?atggtgctgc?caaggagcca?ccgccacctt?acgtgtctgc?ctaa 954
<210>2
<211>1440
<212>DNA
＜213〉Genus Homo people (Homo sapiens)
<400>2
gccgactagg?ggactggcgg?agggtgcacg?ctgatggatt?tactcaccgg?gtgcttggag 60
ctccagcagc?tggctggagc?ccgcgatgac?gtcacggact?cgggtcacat?ggccgagtcc 120
gccccgcccc?ctccccgtcc?ccgccgctgc?agccgtcgcc?ttcggagcga?agggtaccga 180
cccggcagaa?gctcggagct?ctcggggtat?cgaggaggca?ggcccgcggg?cgcacgggcg 240
agcgggccgg?gagccggagc?ggcggaggag?ccggcagcag?cggcgcggcg?ggctccaggc 300
gaggcggtcg?acgctcctga?aaacttgcgc?gcgcgctcgc?gccactgcgc?ccggagcgat 360
gaagatggtc?gcgccctgga?cgcggttcta?ctccaacagc?tgctgcttgt?gctgccatgt 420
ccgcaccggc?accatcctgc?tcggcgtctg?gtatctgatc?atcaatgctg?tggtactgtt 480
gattttattg?agtgccctgg?ctgatccgga?tcagtataac?ttttcaagtt?ctgaactggg 540
aggtgacttt?gagttcatgg?atgatgccaa?catgtgcatt?gccattgcga?tttctcttct 600
catgatcctg?atatgtgcta?tggctactta?cggagcgtac?aagcaacgcg?cagcctggat 660
catcccattc?ttctgttacc?agatctttga?ctttgccctg?aacatgttgg?ttgcaatcac 720
tgtgcttatt?tatccaaact?ccattcagga?atacatacgg?caactgcctc?ctaattttcc 780
ctacagagat?gatgtcatgt?cagtgaatcc?tacctgtttg?gtccttatta?ttcttctgtt 840
tattagcatt?atcttgactt?ttaagggtta?cttgattagc?tgtgtttgga?actgctaccg 900
atacatcaat?ggtaggaact?cctctgatgt?cctggtttat?gttaccagca?atgacactac 960
ggtgctgcta?cccccgtatg?atgatgccac?tgtgaatggt?gctgccaagg?agccaccgcc 1020
accttacgtg?tctgcctaag?ccttcaagtg?ggcggagctg?agggcagcag?cttgactttg 1080
cagacatctg?agcaatagtt?ctgttatttc?acttttgcca?tgagcctctc?tgagcttgtt 1140
tgttgctgaa?atgctacttt?ttaaaattta?gatgttagat?tgaaaactgt?agttttcaac 1200
atatgctttg?ctggaacact?gtgatagatt?aactgtagaa?ttcttcctgt?acgattgggg 1260
atataatggg?cttcactaac?cttccctagg?cattgaaact?tcccccaaat?ctgatggacc 1320
tagaagtctg?cttttgtacc?tgctgggccc?caaagttggg?catttttctc?tctgttccct 1380
ctcttttgaa?aatgtaaaat?aaaaccaaaa?atagaccaaa?aaaaaaaaaa?aaaaaaaaaa 1440
<210>3
<211>2169
<212>DNA
＜213〉Genus Homo people (Homo sapiens)
<400>3
gccgactagg?ggactggcgg?agggtgcacg?ctgatggatt?tactcaccgg?gtgcttggag 60
ctccagcagc?tggctggagc?ccgcgatgac?gtcacggact?cgggtcacat?ggccgagtcc 120
gccccgcccc?ctccccgtcc?ccgccgctgc?agccgtcgcc?ttcggagcga?agggtaccga 180
cccggcagaa?gctcggagct?ctcggggtat?cgaggaggca?ggcccgcggg?cgcacgggcg 240
agcgggccgg?gagccggagc?ggcggaggag?ccggcagcag?cggcgcggcg?ggctccaggc 300
gaggcggtcg?acgctcctga?aaacttgcgc?gcgcgctcgc?gccactgcgc?ccggagcgat 360
gaagatggtc?gcgccctgga?cgcggttcta?ctccaacagc?tgctgcttgt?gctgccatgt 420
ccgcaccggc?accatcctgc?tcggcgtctg?gtatctgatc?atcaatgctg?tggtactgtt 480
gattttattg?agtgccctgg?ctgatccgga?tcagtataac?ttttcaagtt?ctgaactggg 540
aggtgacttt?gagttcatgg?atgatgccaa?catgtgcatt?gccattgcga?tttctcttct 600
catgatcctg?atatgtgcta?tggctactta?cggagcgtac?aagcaacgcg?cagcctggat 660
catcccattc?ttctgttacc?agatctttga?ctttgccctg?aacatgttgg?ttgcaatcac 720
tgtgcttatt?tatccaaact?ccattcagga?atacatacgg?caactgcctc?ctaattttcc 780
ctacagagat?gatgtcatgt?cagtgaatcc?tacctgtttg?gtccttatta?ttcttctgtt 840
tattagcatt?atcttgactt?ttaagggtta?cttgattagc?tgtgtttgga?actgctaccg 900
atacatcaat?ggtaggaact?cctctgatgt?cctggtttat?gttaccagca?atgacactac 960
ggtgctgcta?cccccgtatg?atgatgccac?tgtgaatggt?gctgccaagg?agccaccgcc 1020
accttacgtg?tctgcctaag?ccttcaagtg?ggcggagctg?agggcagcag?cttgactttg 1080
cagacatctg?agcaatagtt?ctgttatttc?acttttgcca?tgagcctctc?tgagcttgtt 1240
tgttgctgaa?atgctacttt?ttaaaattta?gatgttagat?tgaaaactgt?agttttcaac 1200
atatgctttg?ctggaacact?gtgatagatt?aactgtagaa?ttcttcctgt?acgattgggg 1260
atataatggg?cttcactaac?cttccctagg?cattgaaact?tcccccaaat?ctgatggacc 1320
tagaagtctg?cttttgtacc?tgctgggccc?caaagttggg?catttttctc?tctgttccct 1380
ctcttttgaa?aatgtaaaat?aaaaccaaaa?atagacaact?ttttcttcag?ccattccagc 1440
atagagaaca?aaaccttatg?gaaacaggaa?tgtcaattgt?gtaatcattg?ttctaattag 1500
gtaaatagaa?gtccttatgt?atgtgttaca?agaatttccc?ccacaacatc?ctttatgact 1560
gaagttcaat?gacagtttgt?gtttggtggt?aaaggatttt?ctccatggcc?tgaattaaga 1620
ccattagaaa?gcaccaggcc?gtgggagcag?tgaccatctg?ctgactgttc?ttgtggatct 1680
tgtgtccagg?gacatggggt?gacatgcctc?gtatgtgtta?gagggtggaa?tggatgtgtt 1740
tggcgctgca?tgggatctgg?tgcccctctt?ctcctggatt?cacatcccca?cccagggccc 1800
gcttttacta?agtgttctgc?cctagattgg?ttcaaggagg?tcatccaact?gactttatcg 1860
agtggaattg?ggatatattt?gatatacttc?tgcctaacaa?catggaaaag?ggttttcttt 1920
tccctgcaag?ctacatccta?ctgctttgaa?cttccaagta?tgtctagtca?ccttttaaaa 1980
tgtaaacatt?ttcagaaaaa?tgaggattgc?cttccttgta?tgcgcttttt?accttgacta 2040
cctgaattgc?aagggatttt?tatatattca?tatgttacaa?agtcagcaac?tctcctgttg 2100
gttcattatt?gaatgtgctg?taaattaagt?tgtttgcaat?taaaacaagg?tttgcccaca 2160
aaaaaaaaa 2169
<210>4
<211>317
<212>PRT
＜213〉Genus Homo people (Homo sapiens)
<400>4
Met?Thr?Ser?Arg?Thr?Arg?Val?Thr?Trp?Pro?Ser?Pro?Pro?Arg?Pro
1 5 10 15
Leu?Pro?Val?Pro?Ala?A1a?Ala?Ala?Val?Ala?Phe?Gly?Ala?Lys?Gly
20 25 30
Thr?Asp?Pro?Ala?Glu?Ala?Arg?Ser?Ser?Arg?Gly?Ile?Glu?Glu?Ala
35 40 45
Gly?Pro?Arg?Ala?His?Gly?Arg?Ala?Gly?Arg?Glu?Pro?Glu?Arg?Arg
50 55 60
Arg?Ser?Arg?Gln?Gln?Arg?Arg?Gly?Gly?Leu?Gln?Ala?Arg?Arg?Ser
65 70 75
Thr?Leu?Leu?Lys?Thr?Cys?Ala?Arg?Ala?Arg?Ala?Thr?Ala?Pro?Gly
80 85 90
Ala?Met?Lys?Met?Val?Ala?Pro?Trp?Thr?Arg?Phe?Tyr?Ser?Asn?Ser
95 100 105
Cys?Cys?Leu?Cys?Cys?His?Val?Arg?Thr?Gly?Thr?Ile?Leu?Leu?Gly
110 115 120
Val?Trp?Tyr?Leu?Ile?Ile?Asn?Ala?Val?Val?Leu?Leu?Ile?Leu?Leu
125 130 135
Ser?Ala?Leu?Ala?Asp?Pro?Asp?Gln?Tyr?Asn?Phe?Ser?Ser?Ser?Glu
140 145 150
Leu?Gly?Gly?Asp?Phe?Glu?Phe?Met?Asp?Asp?Ala?Asn?Met?Cys?Ile
155 160 165
Ala?Ile?Ala?Ile?Ser?Leu?Leu?Met?Ile?Leu?Ile?Cys?Ala?Met?Ala
170 175 180
Thr?Tyr?Gly?Ala?Tyr?Lys?Gln?Arg?Ala?Ala?Trp?Ile?Ile?Pro?Phe
185 190 195
Phe?Cys?Tyr?Gln?Ile?Phe?Asp?Phe?Ala?Leu?Asn?Met?Leu?Val?Ala
200 205 210
Ile?Thr?Val?Leu?Ile?Tyr?Pro?Asn?Ser?I1e?Gln?Glu?Tyr?Ile?Arg
215 220 225
Gln?Leu?Pro?Pro?Asn?Phe?Pro?Tyr?Arg?Asp?Asp?Val?Met?Ser?Val
230 235 240
Asn?Pro?Thr?Cys?Leu?Val?Leu?Ile?Ile?Leu?Leu?Phe?Ile?Ser?Ile
245 250 255
Ile?Leu?Thr?Phe?Lys?Gly?Tyr?Leu?Ile?Ser?Cys?Val?Trp?Asn?Cys
260 265 270
Tyr?Arg?Tyr?Ile?Asn?Gly?Arg?Asn?Ser?Ser?Asp?Val?Leu?Val?Tyr
275 280 285
Val?Thr?Ser?Asn?Asp?Thr?Thr?Val?Leu?Leu?Pro?Pro?Tyr?Asp?Asp
290 295 300
Ala?Thr?Val?Asn?Gly?Ala?Ala?Lys?Glu?Pro?Pro?Pro?Pro?Tyr?Val
305 310 315
Ser?Ala
317
<210>5
<211>226
<212>PRT
＜213〉Genus Homo people (Homo sapiens)
<400>5
Met?Lys?Met?Val?Ala?Pro?Trp?Thr?Arg?Phe?Tyr?Ser?Asn?Ser?Cys
1 5 10 15
Cys?Leu?Cys?Cys?His?Val?Arg?Thr?Gly?Thr?Ile?Leu?Leu?Gly?Val
20 25 30
Trp?Tyr?Leu?Ile?Ile?Asn?Ala?Val?Val?Leu?Leu?Ile?Leu?Leu?Ser
35 40 45
Ala?Leu?Ala?Asp?Pro?Asp?Gln?Tyr?Asn?Phe?Ser?Ser?Ser?Glu?Leu
50 55 60
Gly?Gly?Asp?Phe?Glu?Phe?Met?Asp?Asp?Ala?Ash?Met?Cys?Ile?Ala
65 70 75
Ile?Ala?Ile?Ser?Leu?Leu?Met?Ile?Leu?Ile?Cys?Ala?Met?Ala?Thr
80 85 90
Tyr?Gly?Ala?Tyr?Lys?Gln?Arg?Ala?Ala?Trp?Ile?Ile?Pro?Phe?Phe
95 100 105
Cys?Tyr?Gln?Ile?Phe?Asp?Phe?Ala?Leu?Asn?Met?Leu?Val?Ala?Ile
110 115 120
Thr?Val?Leu?Ile?Tyr?Pro?Asn?Ser?Ile?Gln?Glu?Tyr?Ile?Arg?Gln
125 130 135
Leu?Pro?Pro?Asn?Phe?Pro?Tyr?Arg?Asp?Asp?Val?Met?Ser?Val?Asn
140 145 150
Pro?Thr?Cys?Leu?Val?Leu?Ile?Ile?Leu?Leu?Phe?Ile?Ser?Ile?Ile
155 160 165
Leu?Thr?Phe?Lys?Gly?Tyr?Leu?Ile?Ser?Cys?Val?Trp?Asn?Cys?Tyr
170 175 180
Arg?Tyr?Ile?Asn?Gly?Arg?Asn?Ser?Ser?Asp?Val?Leu?Val?Tyr?Val
185 190 195
Thr?Ser?Asn?Asp?Thr?Thr?Val?Leu?Leu?Pro?Pro?Tyr?Asp?Asp?Ala
200 205 210
Thr?Val?Asn?Gly?Ala?Ala?Lys?Glu?Pro?Pro?Pro?Pro?Tyr?Val?Ser
215 220 225
Ala
226
Claims (8)
1, people's liver cancer related gene LAPTM4B is one of following nucleotide sequences:
1) the SEQ ID № in the sequence table: 1;
2) SEQ ID № in the code sequence tabulation: 4 or SEQ ID №: the polynucleotide of 5 protein sequences.
2, people's liver cancer related gene LAPTM4B according to claim 1 is characterized in that: described gene is the SEQ ID № in the sequence table: 1.
3, people's liver cancer related gene LAPTM4B according to claim 2 is characterized in that: described gene is the SEQ ID № in the sequence table: 2.
4, people's liver cancer related gene LAPTM4B according to claim 2 is characterized in that: described gene is the SEQ ID № in the sequence table: 3.
5, human hepatoma associated protein LAPTM4B, be to have sequence 4 in the sequence table or/and the protein of sequence 5 aminoacid sequences, or with sequence 4 or/and the aminoacid sequence of sequence 5 through replacement, disappearance or the interpolation of one or several amino-acid residue and have with sequence 4 or/and the aminoacid sequence of sequence 5 identical active by sequence 4 or/and sequence 5 deutero-protein.
6, human hepatoma associated protein LAPTM4B according to claim 5 is characterized in that: it is to have sequence 4 in the sequence table or/and the protein of sequence 5 aminoacid sequences.
7, contain the described expression carrier of claim 1.
8, the transgenic cell line that contains the described gene of claim 1.
Priority Applications (11)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 02158110 CN1239700C (en) | 2002-12-24 | 2002-12-24 | Human liver cancer related gene, encoded product and use thereof |
| AU2003292867A AU2003292867A1 (en) | 2002-12-24 | 2003-12-24 | Human cancer-relating genes, the products encoded thereby and applications thereof |
| EP03782072A EP1577390B9 (en) | 2002-12-24 | 2003-12-24 | Human cancer-related genes, the products encoded thereby and applications thereof |
| EP10153116.8A EP2230306B1 (en) | 2002-12-24 | 2003-12-24 | Human cancer-relating genes, its encoded products and applications |
| US10/540,539 US7910711B2 (en) | 2002-12-24 | 2003-12-24 | Human cancer-relating genes, the products encoded thereby and applications thereof |
| JP2005509681A JP4617257B2 (en) | 2002-12-24 | 2003-12-24 | Human cancer-related genes, products encoded by them and applications |
| PCT/CN2003/001109 WO2004058971A1 (en) | 2002-12-24 | 2003-12-24 | Human cancer-relating genes, the products encoded thereby and applications thereof |
| DE60336015T DE60336015D1 (en) | 2002-12-24 | 2003-12-24 | Human genes associated with cancer, including encoded products and applications thereof |
| AT03782072T ATE498012T1 (en) | 2002-12-24 | 2003-12-24 | HUMAN GENES RELATED TO CANCER, PRODUCTS ENCODED THEREOF, AND APPLICATIONS THEREOF |
| US13/019,297 US20120148589A1 (en) | 2002-12-24 | 2011-02-01 | Human cancer-related gene, its encoded products and applications |
| US15/146,307 US9470690B2 (en) | 2002-12-24 | 2016-05-04 | Human cancer-related gene, its encoded products and applications |
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| CN 02158110 CN1239700C (en) | 2002-12-24 | 2002-12-24 | Human liver cancer related gene, encoded product and use thereof |
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| CN1510138A CN1510138A (en) | 2004-07-07 |
| CN1239700C true CN1239700C (en) | 2006-02-01 |
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| CN101393217B (en) * | 2008-09-27 | 2012-05-30 | 中国人民解放军第二军医大学 | Detection kit for schistosomiasis japonica blood serum designated object |
| CN101781357B (en) * | 2009-01-21 | 2012-01-11 | 中国科学院化学研究所 | Polypeptide for detecting human cancer cells, and application thereof |
| CN117783529B (en) * | 2024-01-02 | 2024-09-13 | 复旦大学 | Application of reagent for detecting N-glycosylation modified protein in preparation of liver cancer diagnosis product |
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