Abstract: This invention provides a method of generating a subtracted cDNA library of a cell comprising: a) generating a cDNA library of the cell; b) isolating double-stranded DNAs from the cDNA library; c) releasing the double-stranded cDNA inserts from the double-stranded DNAs; d) denaturing the isolated double-stranded cDNA inserts; e) hybridizing the denatured double-stranded cDNA inserts with a labelled single-stranded nucleic acid molecules which are to be subtracted from the cDNA library; and f) separating the hybridized labeled single-stranded nucleic acid molecule from the double-stranded cDNA inserts, thereby generating a subtracted cDNA library of a cell. This invention also provides different uses of the subtracted library.

Abstract: This invention provides a vector suitable for introduction into a cell, having: a) an inducible PEG-3 regulatory region; and b) a gene encoding a product that causes or may be induced to cause the death or inhibition of cancer cell growth. In addition, this invention further provides the above-described vectors, wherein the inducible PEG-3 regulatory region is a promoter. This invention further provides the above-described vectors, wherein the gene encodes an inducer of apoptosis. In addition, this invention provides the above-described vectors, wherein the gene is a tumor suppressor gene. In addition, this invention provides the above-described vectors, wherein the gene encodes a viral replication protein. This invention also provides the above-described vectors, wherein the gene encodes a product toxic to cells or an intermediate to a product toxic to cells. In addition, this invention provides the above-described vectors, wherein the gene encodes a product causing enhanced immune recognition of the cell.

Abstract: This invention provides a method for detecting cancer cells in a sample comprising detection of the expression of a Prostate Tumor Inducing Gene in the sample, wherein a positive detection of the expression indicates the presence of cancer cells in the sample. In an embodiment, the expression is performed by measuring the level of PTI mRNA. The mRNA is measured by reverse transcription polymerase chain reaction using at least a pair of appropriate primers. It is well known in the art that once the sequence of the Prostate Tumor Inducing Gene is determined, appropriate pair of primers may be selected. This invention also provides the above method, wherein the expression is performed by measuring the level of PTI protein.

Abstract: A vector having the Protein Translation Peptide Elongation Factor-1&agr; (EF-1&agr;) promoter and nucleic acids encoding a reverse tetracycline-controlled activator is provided, wherein the expression of the activator is under the control of the EF-1&agr; promoter. In addition, a method of generating a reverse tetracycline-controlled transactivator expression system for inducible tetracycline-regulated gene expression is provided that consists of: (a) isolation of a DNA fragment encoding the reverse tetracycline-controlled transactivator, (b) isolation of a DNA fragment containing the EF-1&agr; promoter, (c) subcloning of these DNA fragments into a plasmid or other suitable vector to create a vector in which the reverse tetracycline-controlled transactivator is operably linked to the EF-1&agr; promoter.

Abstract: This invention provides a method for reversing the cancerous phenotype of a cancer cell by introducing a nucleic acid having the melanoma differentiation associated gene (mda-7) into the cell under conditions that permit the expression of the gene so as to thereby reverse the cancerous phenotype of the cell. This invention also provides a method for reversing the cancerous phenotype of a cancer cell by introducing the gene product of the above-described gene into the cancerous cell so as to thereby reverse the cancerous phenotype of the cell. This invention also provides a pharmaceutical composition having the melanoma differentiation associated gene (mda-7) or the gene product of the melanoma differentiation associated gene (mda-7) effective to reverse the cancerous phenotype of a cancer cell and a pharmaceutically acceptable carrier.

Abstract: This invention provides a method for reversing cancer phenotype of a cancer cell which comprises introducing an exogenous material which is capable of specifically recognizing either a Prostate Tumor Inducing Gene, RNA of said gene or gene product of said gene into the cell under conditions permitting inhibition of the expression of said gene or function of said gene product so as to thereby reverse the cancerous phenotype of the cell. This invention also provides a method for reversing cancer phenotype of a cancer cell in a subject which comprises introducing an exogenous material which is capable of specifically recognizing a Prostate Tumor Inducing Gene, RNA of said gene or gene product of said gene into the subject's cancer cell under conditions permitting inhibition of the expression of said gene or function of said gene product in the subject's cell so as to thereby reverse the cancerous phenotype of the cell.

Abstract: Methods are provided for identifying agents that modulate the expression of genes, such as genes encoding tumor suppressor or tumor-promoting proteins. The methods generally comprise screening candidate agents for the ability to enhance expression of a tumor suppressor gene, such as mda-7, or for the ability to inhibit expression of a tumor-promoting gene within a cell. Modulating agents may be used, for example, within anti-cancer therapies.

Abstract: This invention providing a method of producing a sequential cDNA library array comprising (a) obtaining a subtracted cDNA library having clones containing cDNA inserts and said library also allows propagation of the cDNA inserts (b) arraying the cDNA library into individual clone such that each clone can propagate on its own (c) transferring the arrayed clones onto a solid matrix thereby generating a cDNA library replica (d) extracting the nucleic acid from the clone and (e) generating a subarray of phage clones or DNA inserts. This invention also provides a method of identifying genes comprising hybridizing the cDNA library array with specific probes. The invention also provides the genes identified by the preceeding method. This invention provides a method wherein the sequential cDNA library array is used for diagnostics, genetic screening and prognosis.

Abstract: A composition for the upregulation of expression of cell antigens, without inducing shedding, which comprises a protein kinase C activator is provided for this invention. Further provided by this invention is a method of detecting and treating tumor cells comprising contacting tumor cells with an effective amount of a protein kinase C activator for the upregulation of expression of antigens of tumor cells, without inducing antigen shedding, and detecting the presence of said antigen or then further contacting said tumor cells with an effective amount of an antibody directed to said antigen.

Abstract: This invention provides a method for reversing the cancerous phenotype of a cancer cell by introducing a nucleic acid having the melanoma differentiation associated gene (mda-7) into the cell under conditions that permit the expression of the gene so as to thereby reverse the cancerous phenotype of the cell. This invention also provides a method for reversing the cancerous phenotype of a cancer cell by introducing the gene product of the above-described gene into the cancerous cell so as to thereby reverse the cancerous phenotype of the cell. This invention also provides a pharmaceutical composition having an amount of a nucleic acid having the melanoma differentiation associated gene (mda-7) or the gene product of a melanoma differentiation associated gene (mda-7) effective to reverse the cancerous phenotype of a cancer cell and a pharmaceutically acceptable carrier.

Abstract: The present invention provides a vector comprising Protein Translation Peptide Elongation Factor-1 &agr; promoter and nucleic acids encoding reverse tetracycline controlled transactivator, wherein the expression of said transactivator is under the control of Protein Translation Peptide Elongation Factor-1 &agr; promoter.

Abstract: The present invention provides a general method for identifying genes and producing immunological reagents which encode cell surface antigens of human origin. Specifically, this invention provides a method for preparing a hybridoma cell line which produces an antibody which specifically recognizes and binds to a cell surface antigen associated with a neoplastic, human cell. This invention also provides a method for preparing a monoclonal and a polyclonal antibody which specifically recognizes and binds to a cell surface antigen associated with a neoplastic, human cell. Using the antibodies produced, this invention further provides a method for diagnosing in a subject a neoplastic condition, a method for treating a neoplastic condition, and a method for imaging a neoplastic, human cell.

Abstract: This invention provides a vector suitable for introduction into a cell, comprising: a) an inducible PEG-3 regulatory region; and b) a gene encoding a product that causes or may be induced to cause the death or inhibition of cancer cell growth. In addition, this invention further provides the above-described vectors, wherein the inducible PEG-3 regulatory region is a promoter. This invention further provides the above-described vectors, wherein the gene encodes an inducer of apoptosis. In addition, this invention provides the above-described vectors, wherein the gene is a tumor suppressor gene. In addition, this invention provides the above-described vectors, wherein the gene encodes a viral replication protein. This invention also provides the above-described vectors, wherein the gene encodes a product toxic to cells or an intermediate to a product toxic to cells.

Abstract: This invention provides a method for determining whether a compound is capable of suppressing ras functions comprising: (a) contacting an effective amount of the compound with Ha-ras transformed cloned rat embryo fibroblast cells under conditions permitting the compound to suppress ras functions in the cells; and (b) determining the expression or inhibition of certain indicator gene or genes, thereby determining whether the compound is capable of suppressing ras function. This invention further provides the determined compound and a pharmaceutical composition comprising the compound and a pharmaceutically acceptable carrier. This invention also provides methods for generating transcriptional switched Ha-ras transformed cloned rat embryo fibroblast cells. This invention also provides the generated cells and different uses of the cells.

Abstract: This invention provides a method of detecting cancer metastatic cells in a subject, comprising: a) obtaining a nucleic acid sample from the subject's blood; b) amplifying nucleic acid encoding the product of prostate carcinoma tumor antigen gene-1; and c) detecting the presence of nucleic acid encoding the product of the prostate carcinoma tumor antigen gene-1, thereby detecting cancer metastatic cells in a subject. This invention also provides the above-described methods, wherein the method of amplification is PCR. This invention further provides the above-described methods, wherein the primers are 5′-AAGCTGACGCCTCATTTGCA-3′ SEQ ID NO: 1 and 5′-AACCACCAATGGAACTGGGT-3′ SEQ ID NO: 2. This invention also provides the above-described methods, wherein the primers are 5′-AATGGCTTCTGTGATACT-3′ SEQ ID NO: 3 and 5′-GGCTATAAGTGTTGCTGC-3′SEQ ID NO: 4.

Abstract: This invention provides a method of identifying a tumor suppressor gene of a cell(s). Analogous methods to identify tumor suppressors in normal cells and to identify genes associated with unknown genetic defects are also described. The feasibility of the title method was demonstrated by observing the effects of caffeine acid phenethyl ester on oncogene-tranformed CREF cells. In a second series of expts., human papillomavirus 18-transformed CREF cells were transfected with human fibroblast cDNA cloned into a pMAM-neo vector which allows dexamethasone-inducible expression. After growth in the presence of G418, neomycin resistant transformed CREF cells were obsd. Application of dexamethasone resulted in appearance of cells with normal phenotype.

Abstract: This invention provides a method for determining whether a compound is capable of suppressing ras functions comprising: (a) contacting an effective amount of the compound with Ha-ras transformed cloned rat embryo fibroblast cells under conditions permitting the compound to suppress ras functions in the cells; and (b) determining the expression or inhibition of certain indicator gene or genes, thereby determining whether the compound is capable of suppressing ras function. This invention further provides the determined compound and a pharmaceutical composition comprising the compound and a pharmaceutically acceptable carrier. This invention also provides methods for generating transcriptional switched Ha-ras transformed cloned rat embryo fibroblast cells. This invention also provides the generated cells and different uses of the cells.

Abstract: A composition for the upregulation of expression of cell antigens, without inducing shedding, which comprises a protein kinase C activator is provided by this invention. Further provided by this invention is a method of detecting and treating tumor cells comprising contacting tumor cells with an effective amount of a protein kinase C activator for the upregulation of expression of antigens of tumor cells, without inducing antigen shedding, and detecting the presence of said antigen or then further contacting said tumor cells with an effective amount of an antibody directed to said antigen.

Abstract: This invention provides a CREF-Trans 6 cell line. The invention further provides CREF-Trans 6 cell lines containing DNA from an established cancer cell line or a primary tumor. This invention also provides the use of said cell lines to obtain DNA encoding a cell surface antigen associated with a neoplastic human cell line.

Abstract: This invention provides a method for preparing a hybridoma cell line which produces a monoclonal antibody which specifically recognizes and binds to a tumor associated antigen which comprises: (a) cotransfecting a CREF-Trans 6 cell line with DNA isolated from a neoplastic, human cell and a plasmid which encodes a selectable or identifiable trait; (b) selecting transfected cells which express the selectable or identifiable trait; (c) recovering the cells so selected in step (b); (d) injecting the cells so recovered in step (c) into a suitable marine host; (e) maintaining the resulting first murine host for a period of time effective to induce the cells injected in step (d) to form a tumor in the murine host; (f) isolating the tumor formed in step (e); (g) obtaining tumor cells from the isolated tumor in step (f); (h) coating the tumor cells obtained in step (9) with an antiserum generated against the CREF Trans-6 cell line (i) injecting the antiserum-coated cells from step (h) into a plurality of suitable seco