WO1997012975B1 - An epithelial protein and dna thereof for use in early cancer detection - Google Patents
An epithelial protein and dna thereof for use in early cancer detectionInfo
- Publication number
- WO1997012975B1 WO1997012975B1 PCT/US1996/015825 US9615825W WO9712975B1 WO 1997012975 B1 WO1997012975 B1 WO 1997012975B1 US 9615825 W US9615825 W US 9615825W WO 9712975 B1 WO9712975 B1 WO 9712975B1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- probe
- epithelial
- labeled
- nucleic acid
- protein
- Prior art date
Links
- 206010028980 Neoplasm Diseases 0.000 title claims abstract 4
- 201000011510 cancer Diseases 0.000 title claims abstract 4
- 108090000623 proteins and genes Proteins 0.000 title abstract 10
- 102000004169 proteins and genes Human genes 0.000 title abstract 10
- 238000001514 detection method Methods 0.000 title abstract 3
- 238000000034 method Methods 0.000 claims abstract 11
- 108010019372 Heterogeneous-Nuclear Ribonucleoproteins Proteins 0.000 claims abstract 6
- 102000006479 Heterogeneous-Nuclear Ribonucleoproteins Human genes 0.000 claims abstract 6
- 108020004999 messenger RNA Proteins 0.000 claims abstract 3
- 239000000523 sample Substances 0.000 claims 16
- 108091028043 Nucleic acid sequence Proteins 0.000 claims 6
- 150000007523 nucleic acids Chemical group 0.000 claims 6
- SHIBSTMRCDJXLN-UHFFFAOYSA-N Digoxigenin Natural products C1CC(C2C(C3(C)CCC(O)CC3CC2)CC2O)(O)C2(C)C1C1=CC(=O)OC1 SHIBSTMRCDJXLN-UHFFFAOYSA-N 0.000 claims 2
- QONQRTHLHBTMGP-UHFFFAOYSA-N digitoxigenin Natural products CC12CCC(C3(CCC(O)CC3CC3)C)C3C11OC1CC2C1=CC(=O)OC1 QONQRTHLHBTMGP-UHFFFAOYSA-N 0.000 claims 2
- SHIBSTMRCDJXLN-KCZCNTNESA-N digoxigenin Chemical compound C1([C@@H]2[C@@]3([C@@](CC2)(O)[C@H]2[C@@H]([C@@]4(C)CC[C@H](O)C[C@H]4CC2)C[C@H]3O)C)=CC(=O)OC1 SHIBSTMRCDJXLN-KCZCNTNESA-N 0.000 claims 2
- 230000003287 optical effect Effects 0.000 claims 2
- 108091032973 (ribonucleotides)n+m Proteins 0.000 claims 1
- 108020004711 Nucleic Acid Probes Proteins 0.000 claims 1
- 239000002853 nucleic acid probe Substances 0.000 claims 1
- 239000002773 nucleotide Substances 0.000 claims 1
- 125000003729 nucleotide group Chemical group 0.000 claims 1
- 206010058467 Lung neoplasm malignant Diseases 0.000 abstract 3
- 201000005202 lung cancer Diseases 0.000 abstract 3
- 208000020816 lung neoplasm Diseases 0.000 abstract 3
- 210000004027 cell Anatomy 0.000 abstract 2
- 108090000765 processed proteins & peptides Proteins 0.000 abstract 2
- 238000000746 purification Methods 0.000 abstract 2
- 208000005623 Carcinogenesis Diseases 0.000 abstract 1
- 238000011537 Coomassie blue staining Methods 0.000 abstract 1
- 241000124008 Mammalia Species 0.000 abstract 1
- 206010035226 Plasma cell myeloma Diseases 0.000 abstract 1
- 238000005571 anion exchange chromatography Methods 0.000 abstract 1
- 210000000988 bone and bone Anatomy 0.000 abstract 1
- 210000000481 breast Anatomy 0.000 abstract 1
- 210000000424 bronchial epithelial cell Anatomy 0.000 abstract 1
- 230000036952 cancer formation Effects 0.000 abstract 1
- 231100000504 carcinogenesis Toxicity 0.000 abstract 1
- 238000004113 cell culture Methods 0.000 abstract 1
- 210000002919 epithelial cell Anatomy 0.000 abstract 1
- 108010025934 hnRNP A2 Proteins 0.000 abstract 1
- 230000001900 immune effect Effects 0.000 abstract 1
- 238000011532 immunohistochemical staining Methods 0.000 abstract 1
- 238000012744 immunostaining Methods 0.000 abstract 1
- 238000001155 isoelectric focusing Methods 0.000 abstract 1
- 210000003734 kidney Anatomy 0.000 abstract 1
- 210000004072 lung Anatomy 0.000 abstract 1
- 239000003550 marker Substances 0.000 abstract 1
- 201000001441 melanoma Diseases 0.000 abstract 1
- 238000007479 molecular analysis Methods 0.000 abstract 1
- 238000012544 monitoring process Methods 0.000 abstract 1
- 201000000050 myeloid neoplasm Diseases 0.000 abstract 1
- 210000001672 ovary Anatomy 0.000 abstract 1
- 229920000642 polymer Polymers 0.000 abstract 1
- 102000004196 processed proteins & peptides Human genes 0.000 abstract 1
- 210000002307 prostate Anatomy 0.000 abstract 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 abstract 1
- 230000009466 transformation Effects 0.000 abstract 1
- 238000001262 western blot Methods 0.000 abstract 1
Abstract
The present invention is a purified and isolated epithelial protein, peptide and variants thereof whose increased presence in an epithelial cell is indicative of precancer. One epithelial protein which is an early detection marked for lung cancer was purified from two human lung cancer cell lines, NCI-H720 and NCI-H157. Using a six-step procedure, the epithelial protein was purified using a Western blot detection system under both non-reducing and reducing conditions. Purification steps included anion exchange chromatography, preparative isoelectric focusing, polymer-based C18HPLC and analytic C4HPLC. After an approximately 25,000 fold purification the immunostaining protein was >90 % pure as judged by coomassie blue staining after reducing SDS-PAGE. The primary epithelial protein share some sequence homology with the heterogeneous nuclear ribonucleoprotein (hnRNP) A2. A minor co-purifying epithelial protein shares some sequence homology with the splice variant hnRNP-B1. Molecular analysis of primary normal bronchial epithelial cell cultures demonstrated a low level the epithelial protein expression, consistent with immunohistochemical staining of clinical samples, and an increased level of expression in most lung cancer cells. The epithelial protein is a marker of epithelial transformation in lung, breast, bone, ovary, prostate, kidney, melanoma and myeloma and may be casual in the process of carcinogenesis. Methods are provided for monitoring the expression of the epithelial protein, peptides and variants using molecular and immunological techniques as a screen for precancer and cancer in mammals. A method of computerized diagnoses of cancer and precancer is provided which detects levels of hnRNP messenger RNA.
Claims
AMENDED CLAIMS
[received by the International Bureau on 21 April 1997 (21.04.97); new claims 59-67 added; remaining claims unchanged (2 pages)]
54. The method according to claim 53 wherein the optical image o is a spatial electronic array.
55. The method according to claim 53 wherein the image is acquired at two different wavelengths.
5
56. The method according to claim 53 wherein the parameter unique to atypical cells is selected from the group consisting of nuclear texture, nuclear ellipse area, optical density, hnRNP mRNA and combinations thereof.
0
57. The method according to claim 53 wherein the cell is treated with a labeled probe, said probe specifically hybridized with hnRNP mRNA.
58. The method according to claim 53 wherein the known 5 negative and known atypical cells are from an archived bank of cells taken from normal humans and humans with cancer or precancer.
59. A labeled nucleic acid sequence probe suitable for use in the o method according to claim 12.
60. A labeled nucleotide probe according to claim 59 comprising a sequence capable of specifically hybridizing with hnRNP or RNA.
5
61. The probes of claim 59 or claim 60, wherein said probe is labeled with digoxigenin.
62. The labeled probe of claim 59 or 60 complexed with a nucleic acid sequence complimentary to the labeled probe.
63. The labeled probe of claim 61 employed with a nucleic acid sequence complimentary to the labeled probe.
-148-
o 64. The labeled probe of claim 62 complexed with a nucleic acid sequence complimentary to the labeled probe, and further comprising an antibody complexed to the label on said probe.
65. The labeled probe of claim 64 wherein said antibody is an 5 anti digoxigenin antibody.
66. The nucleic acid sequence probe of claim 12 or claim 43 complexed with a nucleic acid sequence complimentary to said probe.
67. The nucleic acid probes of claims 59, 60 and 66 having the sequence provided in SEQ ID NO: 11 or SEQ ID NO: 18.
Priority Applications (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| EP96936141A EP0861323A1 (en) | 1995-10-02 | 1996-10-02 | An epithelial protein and dna thereof for use in early cancer detection |
| JP9514401A JP2000500322A (en) | 1995-10-02 | 1996-10-02 | Epithelial proteins and their DNA for use in early cancer detection |
| AU73864/96A AU7386496A (en) | 1995-10-02 | 1996-10-02 | An epithelial protein and dna thereof for use in early cancer detection |
Applications Claiming Priority (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US08/538,711 US5994062A (en) | 1995-10-02 | 1995-10-02 | Epithelial protein and DNA thereof for use in early cancer detection |
| US08/538,711 | 1995-10-02 | ||
| US08/725,027 | 1996-10-02 | ||
| US08/725,027 US6251586B1 (en) | 1995-10-02 | 1996-10-02 | Epithelial protein and DNA thereof for use in early cancer detection |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| WO1997012975A1 WO1997012975A1 (en) | 1997-04-10 |
| WO1997012975B1 true WO1997012975B1 (en) | 1997-05-22 |
Family
ID=27065902
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/US1996/015825 WO1997012975A1 (en) | 1995-10-02 | 1996-10-02 | An epithelial protein and dna thereof for use in early cancer detection |
Country Status (5)
| Country | Link |
|---|---|
| US (1) | US6251586B1 (en) |
| EP (2) | EP1426381A1 (en) |
| JP (1) | JP2000500322A (en) |
| AU (1) | AU7386496A (en) |
| WO (1) | WO1997012975A1 (en) |
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| WO1998014469A2 (en) * | 1996-10-02 | 1998-04-09 | The Government Of The United States Of America As Represented By The Secretary, Department Of Health And Human Services | An epithelial protein and dna thereof for use in early cancer detection |
| US6025470A (en) * | 1997-06-23 | 2000-02-15 | Ludwig Institute For Cancer Research | Isolated nona- and decapeptides which bind to HLA molecules, and the use thereof |
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| GB2354241A (en) * | 1999-09-17 | 2001-03-21 | Rupert Donald Holms | Regulatory/unfolding peptides of ezrin |
| AU3887601A (en) * | 1999-09-22 | 2001-04-24 | Cedars-Sinai Medical Center | Nucleic acids encoding vitamin d response element binding proteins, products related thereto, and methods of using same |
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| US4569788A (en) | 1983-05-18 | 1986-02-11 | The United States Of America As Represented By The Department Of Health And Human Services | Monoclonal antibodies against non small cell lung cancer |
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-
1996
- 1996-10-02 JP JP9514401A patent/JP2000500322A/en active Pending
- 1996-10-02 EP EP03078938A patent/EP1426381A1/en not_active Withdrawn
- 1996-10-02 EP EP96936141A patent/EP0861323A1/en not_active Withdrawn
- 1996-10-02 US US08/725,027 patent/US6251586B1/en not_active Expired - Fee Related
- 1996-10-02 WO PCT/US1996/015825 patent/WO1997012975A1/en not_active Application Discontinuation
- 1996-10-02 AU AU73864/96A patent/AU7386496A/en not_active Abandoned
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