Patent US4442203A successfully refreshed from Google Patents.
US4442203A
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US
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Gene amplification assay for detecting tumor promoters
Assignee: Massachusetts Institute of Technology
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Filing: 1981-06-30
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Granted: 1984-04-10
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Expires: 2001-06-30
Abstract
Abstract A correlation between gene amplification and tumor promotion is disclosed herein. This correlation allows for a simple cellular assay that indicates whether a substance or process is a tumor promoter. This assay does not depend upon numerous biochemical processes that introduce uncontrolled and unascertained varibles into other cellular assays. This assay can also be used to determine whether a set of cells is abnormally genetically labile and therefore susceptible to cancer or genetic disease.
Inventors
- 👤 Alexander J. Varshavsky
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-05 06:02
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/5005; G01N33/5008; G01N33/5014; G01N33/5017; C; C12; C12Q; C12Q1/00; C12Q1/68
Claims Analysis
Total Claims: 16
Claim 1
Independent Claim
50 words
A method for indicating the tumor-promoting characteristics of a substance or process, comprising determining the incidence of gene amplification of a cell culture that is induced by said substance or process and using the induction of gene amplification as an indication of the tumor-promoting capability of the substance or process.
Claim 2
Independent Claim
50 words
A method for indicating the tumor-promoting characteristics of a substance or process, comprising determining the incidence of gene amplification of a cell culture that is induced by said substance or process and using the induction of gene amplification as an indication of the tumor-promoting capability of the substance or process.
Claim 3
Dependent Claim
164 words
A method of claim 1 wherein said incidence of gene amplification is determined by a process comprising the following steps: a. dividing said cell culture into at least one first group of cells and at least one second group of cells; b. contacting said first group with said substance or process; c. contacting said first group and said second group with a second substance that kills cells which do not have a relatively high number of copies of at least one DNA locus that codes for the production of a protein that is the target of said second substance; d. determining the incidence of cells in each of said groups that survive the contact with said second substance; e. comparing the number of surviving cells in each of said groups to determine whether the substance or process affects the incidence of gene amplification; and, f. determining the tumor promoting characteristics of said substance or process from its affect upon the incidence of gene amplification.
Claim 4
Dependent Claim
20 words
A method of claim 2 wherein said second substance is selected from the following group: methotrexate, heavy metals, and phosphonacetyl-L-aspartate.
Claim 5
Independent Claim
160 words
A method for indicating the tumor-promoting characteristics of a substance or process, comprising the following steps: a. dividing said cell culture into at least one first group of cells and at least one second group of cells; b. contacting said first group with said substance or process; c. contacting said first group and said second group with a second substance that kills cells which do not have a relatively high number of copies of at least one DNA locus that codes for the production of a protein that is the target of said second substance; d. determining the incidence of cells in each of said groups that survive the contact with said second substances; e. comparing the number of surviving cells in each of said groups to determine whether the substance or process affects the incidence of gene amplification; and, f. determining the tumor promoting characteristics of said substance or process from its affect upon the incidence of gene amplification.
Claim 6
Independent Claim
160 words
A method for indicating the tumor-promoting characteristics of a substance or process, comprising the following steps: a. dividing said cell culture into at least one first group of cells and at least one second group of cells; b. contacting said first group with said substance or process; c. contacting said first group and said second group with a second substance that kills cells which do not have a relatively high number of copies of at least one DNA locus that codes for the production of a protein that is the target of said second substance; d. determining the incidence of cells in each of said groups that survive the contact with said second substances; e. comparing the number of surviving cells in each of said groups to determine whether the substance or process affects the incidence of gene amplification; and, f. determining the tumor promoting characteristics of said substance or process from its affect upon the incidence of gene amplification.
Claim 7
Dependent Claim
20 words
A method of claim 4 wherein said second substance is selected from the following group: methotrexate, heavy metals, and phosphonacetyl-L-aspartate.
Claim 8
Independent Claim
120 words
A method for indicating whether cells are abnormally genetically labile, comprising the following steps: a. determining the incidence of gene amplification within a culture of said cells that is induced by contact with a first substance or process; b. determining the incidence of gene amplification within a culture of reference cells that are not abnormally genetically labile, that is induced by contact with said first substance; and c. comparing said incidence of induced gene amplification within the culture of cells with the incidence of induced gene amplification within the culture of reference cells to determine whether said cells are abnormally genetically labile, an induction of gene amplification in said cells greater than in the reference cells indicating abnormal genetic lability.
Claim 9
Independent Claim
120 words
A method for indicating whether cells are abnormally genetically labile, comprising the following steps: a. determining the incidence of gene amplification within a culture of said cells that is induced by contact with a first substance or process; b. determining the incidence of gene amplification within a culture of reference cells that are not abnormally genetically labile, that is induced by contact with said first substance; and c. comparing said incidence of induced gene amplification within the culture of cells with the incidence of induced gene amplification within the culture of reference cells to determine whether said cells are abnormally genetically labile, an induction of gene amplification in said cells greater than in the reference cells indicating abnormal genetic lability.
Claim 10
Dependent Claim
141 words
A method of claim 6 wherein the incidence of gene amplification within either culture of cells is determined by a process comprising the following steps: a. dividing the cell culture into at least one first group of cells and at least one second group of cells; b. contacting said first group with a first substance or process that is known to promote tumors or gene amplification; c. contacting said first group and said second group with a second substance that kills cells which do not have a relatively high number of copies of at least one DNA locus that codes for the production of a protein that is the target of said second substance; and d. determining the incidence of cells in each of said groups that survive the contact with said second substance to determine the incidence of gene amplification.
Claim 11
Independent Claim
271 words
A method for indicating whether cells are abnormally genetically labile, comprising the following steps: a. dividing a culture of said first cells into at least one first group and at least one second group; b. contacting said first group with a first substance or process that is known to promote tumors or gene amplification; c. contacting said first group and said second group with a second substance that kills cells which do not have a relatively high number of copies of at least one DNA locus that codes for the production of a protein that is the target of said second substance; d. determining the number of cells in each of said first and second groups that survive the contact with said second substance; e. comparing the incidence of surviving cells in each of said groups to determine the incidence of gene amplification within said cells; f. dividing a popultion of reference cells that are not abnormally genetically labile into at least one third group and at least one forth group; g. contacting said third group with said first substance or process; h. contacting said third group and said fourth group with a said second substance; i. determining the incidence of cells in each of said third and fourth groups that survive contact with said second substance; j. comparing the incidence of surviving cells in each of said groups to determine the incidence of gene amplification within said reference cells; and k. comparing the incidence of gene amplification within said first cells with the incidence of gene amplification within said reference cells to determine whether the cells are abnormal genetically labile.
Claim 12
Independent Claim
271 words
A method for indicating whether cells are abnormally genetically labile, comprising the following steps: a. dividing a culture of said first cells into at least one first group and at least one second group; b. contacting said first group with a first substance or process that is known to promote tumors or gene amplification; c. contacting said first group and said second group with a second substance that kills cells which do not have a relatively high number of copies of at least one DNA locus that codes for the production of a protein that is the target of said second substance; d. determining the number of cells in each of said first and second groups that survive the contact with said second substance; e. comparing the incidence of surviving cells in each of said groups to determine the incidence of gene amplification within said cells; f. dividing a popultion of reference cells that are not abnormally genetically labile into at least one third group and at least one forth group; g. contacting said third group with said first substance or process; h. contacting said third group and said fourth group with a said second substance; i. determining the incidence of cells in each of said third and fourth groups that survive contact with said second substance; j. comparing the incidence of surviving cells in each of said groups to determine the incidence of gene amplification within said reference cells; and k. comparing the incidence of gene amplification within said first cells with the incidence of gene amplification within said reference cells to determine whether the cells are abnormal genetically labile.
Claim 13
Dependent Claim
20 words
A method of claim 8 wherein said second substance is selected from the following group: methotrexate, heavy metals, and phosphonacetyl-L-aspartate.
Claim 14
Independent Claim
115 words
A method for indicating whether cells are abnormally genetically labile, comprising the following steps: a. contacting a culture of said cells with a substance that kills cells which do not have a relatively high number of copies of at least one DNA locus that codes for production of a protein that is the target of said substance; b. contacting a culture of reference cells that are not abnormally genetically labile with said substance; c. determining the incidence of cells in each of said cultures that survive contact with said substance; d. comparing said incidences to determine whether said cells are abnormally genetically labile, a higher rate of survival over the reference cells indicating genetic lability.
Claim 15
Independent Claim
115 words
A method for indicating whether cells are abnormally genetically labile, comprising the following steps: a. contacting a culture of said cells with a substance that kills cells which do not have a relatively high number of copies of at least one DNA locus that codes for production of a protein that is the target of said substance; b. contacting a culture of reference cells that are not abnormally genetically labile with said substance; c. determining the incidence of cells in each of said cultures that survive contact with said substance; d. comparing said incidences to determine whether said cells are abnormally genetically labile, a higher rate of survival over the reference cells indicating genetic lability.
Claim 16
Dependent Claim
19 words
A method of claim 10 wherein said substance is selected from the following group: methotrexate, heavy metals, and phosphonacetyl-L-aspartate.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1987-04-10 | 1987-10-10 | 1988-04-10 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1991-04-10 | 1991-10-10 | 1992-04-10 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1995-04-10 | 1995-10-10 | 1996-04-10 | $7700.00 | Window Closed / Lapsed |