Development and Characterization of a Human Colon Adenocarcinoma Xenograft Deficient in Thymidine Salvage

Peter J. Houghton, Glen Germain, Pamela M. Torrance

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23 Scopus citations

Abstract

In order to determine the contribution of thymidine (dThd) salvage to intrinsic resistance to antimetabolites (5-fluoropyrimidines, antifolates) in the human colon adenocarcinoma xenograft, H×GCj, a subline deficient in thymidine kinase has been developed. A cell line (GC3/M) was established in continuous culture that demonstrated a karyotype identical to that of the stem line of HxGC3(46, X, -Y+12). After inoculation of GC3/M cells into immune-deprived CBA/CaJ mice, the HxGC3/M xenografts retained histological, histochemical, and growth characteristics of the H×GC3xenograft. To develop a line deficient in dThd salvage, GC3/M cells were selected with BrdUrd (100 μg/ml). Three clones characterized were unable to proliferate in HAT medium, and were deficient (<10% control) in the cytosolic form of thymidine kinase. Activities of dThd phosphorylase and dTMP synthase were unchanged from parental GC3/M cells. Of the three clones inoculated into mice, GC3/M TK” 100 C3was tumorigenic, the xenografts demonstrating histological and growth characteristics similar to H×GC3. The in vivo activity of the cytosolic form of dThd kinase was 3.5% of that in H×GC3xenografts. Incorporation in vivo of [methyl-3H]dThd into acid insoluble material was 14% of that in HxGC3tumors. Autoradiographs prepared from these tumors demonstrated that incorporation of radiolabel into nuclei occurred only in stromal cells derived from the host. It is anticipated that H×GC3/M TK-100 C3will be a line valuable for determining the role of dThd salvage in intrinsic resistance to 5-fluoropyrimidines or antifolates in human colon adenocarcinomas growing as xenografts and also the relevance of dTMP synthase as a target for antimetabolites in this histiotype.

Original languageEnglish (US)
Pages (from-to)2117-2122
Number of pages6
JournalCancer Research
Volume47
Issue number8
StatePublished - Apr 1987
Externally publishedYes

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

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