# MECHANISMS REGULATING CELL RESISTANCE TO ADRIAMYCIN

> **NIH NIH R01** · KANSAS STATE UNIVERSITY · 1993 · $115,956

## Abstract

HL60 cells isolated for resistance to adriamycin are multidrug resistant
and defective in the cellular accumulation of drug.  These cells do not
however overexpress mdr1 and do not contain detectable levels of P-
glycoprotein.  Recent evidence indicates that resistance in HL60/Adr cells
is related to overexpression of a new multidrug resistance gene which
encodes a 190 kd (P190) membrane associated ATP binding protein.  A major
focus of the present study will be to molecularly clone and characterize a
cDNA of the P190 gene.  Antibody which is highly reactive with P190 will be
used as a probe in the cDNA cloning procedure.  The nucleotide sequence of
the cDNA will be determined.  If a full length cDNA can be obtained this
material will be placed in an expression vector and used to transfect
sensitive cells.  The properties of the transfected cells will be examined.
The cDNA will also be used to study amplification and expression of the
P190 gene in various resistant isolates and in cells induced to undergo
differentiation.  Additional studies will be carried out to purify P190
using different types of affinity columns and to thereafter determine if
the protein contains ATPase or protein kinase activities.

HL60/Adr cells contain a 150 kd membrane protein (P150) which represents a
hyperphosphorylated form of a protein contained in drug sensitive cells.
In the present study P150 will be partially purified and this material will
be used to prepare a monoclonal antibody against this protein.  The
antibody will be used in immunoprecipitation experiments to further
characterize P150 phosphorylation and the relation of this event to the
development of drug resistance.  Membrane associated protein kinases
involved in the phosphorylation of P150 will isolated and characterized.
The P150 monoclonal antibody will also be used as a probe to clone a P150
cDNA.  This material will be characterized and the nucleotide sequence
determined.

A protein kinase activity capable of phosphorylating P-glycoprotein has
been partially purified from membranes of HL60 cells isolated for
resistance to vincristine.  This enzyme will be further purified and
characterized and its properties with P-glycoprotein as substrate will be
examined in detail.

## Key facts

- **NIH application ID:** 3175367
- **Project number:** 5R01CA037585-10
- **Recipient organization:** KANSAS STATE UNIVERSITY
- **SAM.gov UEI:** CFMMM5JM7HJ9
- **Principal Investigator:** MELVIN S CENTER
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1993
- **Award amount:** $115,956
- **Award type:** 5
- **Project period:** 1984-07-01 → 1995-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3175367

## Citation

> US National Institutes of Health, RePORTER application 3175367, MECHANISMS REGULATING CELL RESISTANCE TO ADRIAMYCIN (5R01CA037585-10). Retrieved via MORVS 2026-10-24 from https://api.morvs.ai/grant/nih/3175367. Licensed CC0.

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