Differential gene expression in peripheral blood lymphocytes of cancer patients treated with whole body hyperthermia and chemotherapy: a pilot study.
OBJECTIVE
The effect of whole body hyperthermia (WBH) at 41.8-42 degrees C on the cellular immune system is still poorly investigated. The aim of this study was to identify genes that become upregulated in peripheral blood lymphocytes (PBLs) of cancer patients during a combined treatment with WBH and chemotherapy by generating complex arrays of cDNA.


METHODS
PBLs were obtained from four patients with different malignancies treated with WBH and varying cytostatic schedules before treatment and immediately thereafter. After constructing subtracted cDNA libraries, clones were screened for cDNA induction by dot-blot and semi-quantitative RT-PCR (sq-RT-PCR).


RESULTS
Among 192 clones, 39 cDNAs were significantly upregulated. Sequencing revealed three groups of genes for which upregulation of mRNA was confirmed by sq-RT-PCR. The first group consisted of genes encoding for various heat shock proteins (HSP 60, 90a, 90b, 105). Further sq-RT-PCR demonstrated differential expression of HSP27 and HSP70 as well. The second group (calcyclin-binding-protein, haemoglobin-beta-chain) comprised genes without pre-specified association to hyperthermia. The cDNA encoding macrophage-inflammatory-protein-1-beta was also observed and may be associated with the pre-described activation of lymphocyte sub-populations during WBH.


CONCLUSIONS
Treatment with WBH and chemotherapy elicits significant short-term effects on the expression of a variety of genes responsible for cellular integrity, stimulation and migration of immune effector cells. Further investigation is warranted to more clearly define the role of those genes for the clinical effect of WBH.
Hildebrandt
Bert
B
Medizinische Klinik mit Schwerpunkt Hämatologie und Onkologie, Campus Virchow-Klinikum, Charité Universitätsmedizin Berlin der Humboldt Universität, Berlin, Germany. bert.hildebrandt@charite.de
Schoeler
Dagmar
D
Ringel
Frauke
F
Kerner
Thoralf
T
Wust
Peter
P
Riess
Hanno
H
Schriever
Folke
F
D000971:Antineoplastic Combined Chemotherapy Protocols / Q000494:pharmacology* / Q000627:therapeutic use; D002135:Calcium-Binding Proteins / Q000097:blood / Q000235:genetics; D054407:Chemokine CCL4; D003131:Combined Modality Therapy; D020869:Gene Expression Profiling; D005914:Globins / Q000235:genetics; D006360:Heat-Shock Proteins / Q000097:blood / Q000235:genetics; D006801:Humans; D006979:Hyperthermia, Induced; D008214:Lymphocytes / Q000378:metabolism*; D019402:Macrophage Inflammatory Proteins / Q000097:blood / Q000235:genetics; D009369:Neoplasms / Q000188:drug therapy; D020411:Oligonucleotide Array Sequence Analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015854:Up-Regulation
C491778:CACYBP protein, human; D002135:Calcium-Binding Proteins; D054407:Chemokine CCL4; D006360:Heat-Shock Proteins; D019402:Macrophage Inflammatory Proteins; D005914:Globins