Project: Research project

Project Details


The objective of this research plan is to generate and
characterize reagents (DNA probes and monoclonal antibodies)
which will ultimately aid in the diagnosis, clinical management
and/or therapy of pancreatic adenocarcinoma. This goal will be
accomplished by identifying and characterizing oncogene activity
previously detected in DNA of the human pancreatic
adenocarcinoma cell line HPAF using the NIH 3T3 transformation
assay. Monoclonal antibodies have previously been raised against
the HPAF transfected NIH 3T3 cells which were selected for
reactivity with HPAF cells and HPAF transfected NIH 3T3 cells
and lack of reactivity with untransfected NIH 3T3 cells. These
will be tested for potential immunodiagnostic uses against a
variety of normal and malignant human tissue specimens. The
biochemical characteristics of the antigens defined by these
antibodies will be established and compared to other known cell
products associated with oncogenes. We will also attempt to
isolate genes coding for the antigens recognized by the
antitransfectant antibodies by constructing and directly screening
an HPAF cDNA expression library using the lambda gt 11 cloning
and expression vector. If the monoclonal antibodies are
unreactive with the cDNA expression library, then second
generation monoclonal antibodies developed against denatured
purified antigen will be used to screen the expression library. If
this procedure was unsuccessful, then a transfectant cDNA library
would be generated which could be positively selected against
HPAF DNA and negatively selected against NIH 3T3 DNA to
identify HPAF positive genes in the transfected cells. Genes
selected in any of the above procedures would be used to select
the intact gene from an HPAF genomic library, which would be
used in transfection experiments to assess transforming
capability. The sequence of these genes would be determined and
used for comparison with other known sequences. Transfection
studies will be repeated using DNA derived from fresh pancreatic
tumor isolates to determine whether the observations made with
the HPAF cell line are applicable to other pancreatic
adenocarcinoma tumors.
Effective start/end date1/1/8712/31/90


  • National Institutes of Health


  • Medicine(all)


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