Multiple drug-resistant human KB carcinoma cells independently selected for high-level resistance to colchicine, adriamycin, or vinblastine show changes in expression of specific proteins.

D W Shen,C Cardarelli,J Hwang,M Cornwell,N Richert,S Ishii,I Pastan,M M Gottesman

doi:10.1016/s0021-9258(19)57466-x

D W Shen, C Cardarelli + Show 6 more

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https://doi.org/10.1016/s0021-9258(19)57466-x

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Abstract

We have established four cell lines derived from the human KB carcinoma cell line which express high-level multiple drug resistance. One of these lines was selected for resistance to colchicine, one was selected for resistance to colchicine in the presence of the tumor promoter, mezerein, one for resistance to vinblastine, and one for resistance to adriamycin. All of these cell lines are cross-resistant to the other selective agents. The development of multidrug resistance in these cultured human carcinoma cells is associated with a limited number of specific protein alterations revealed by high resolution two-dimensional gel electrophoresis and Western blot analysis. These protein alterations in multidrug-resistant lines include the decreased prevalence of members of a family of proteins of molecular mass 70,000 to 80,000 daltons, pI 4.8-5.0, the increased synthesis of a protein of molecular mass 21,000 daltons, pI 5.0, in the colchicine-resistant cell lines only, and the increased expression of a 170,000-dalton protein in membrane preparations from all of the resistant cells. The loss of the 70,000- to 80,000-dalton proteins in the multidrug-resistant lines, which can also be demonstrated by immunoprecipitation of these proteins with specific antisera, is associated with a loss of translatable mRNA for these proteins. These studies suggest that only a limited number of protein changes occur in multidrug-resistant cell lines.

Highlights

We have established four cell lines derived from theChinese hamster ovary (CHO)’ cells [8-12] increased human KB carcinoma cell line which express highgp150,OOO on Chinese hamster lung cells [13] and increased level multiple drug resistance
An dalton proteins in the multidrug-resistalnint es, which analysis of protein synthesis in these cell lines by high resocan be demonstrated by immunoprecipitation of lution two-dimensional gel electrophoresis and Western blot these proteins witshpecific antisera, is associated with analysis shows only a limited number of specific protein a loss of translatable mRNA for these proteins
KB-ADRR5A, the most adriamycin-resistant of the two lines, was chosen for Isolation and Cross-resistance of Multidrug-resistant Cell Lines-We have previously described the selection of colchicine-resistant KB cells in four steps [7]

Summary

Introduction

We have established four cell lines derived from theChinese hamster ovary (CHO)’ cells [8-12] increased human KB carcinoma cell line which express highgp150,OOO on Chinese hamster lung cells [13] and increased level multiple drug resistance. An dalton proteins in the multidrug-resistalnint es, which analysis of protein synthesis in these cell lines by high resocan be demonstrated by immunoprecipitation of lution two-dimensional gel electrophoresis and Western blot these proteins witshpecific antisera, is associated with analysis shows only a limited number of specific protein a loss of translatable mRNA for these proteins. These alterations, some of which are shared by all cell lines and studies suggest that only a limited number of protein some of which are limited to a subset of the multidrugchanges occur in multidrug-resistant cellilnes. We have recently demonstrated the development of multiple drug resistance in a human KB epidermoid carcinoma cell line selected for resistance to colchicine ( 7 )

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