Induction of phenotypic changes in SCLC cell lines in vitro by hexamethylene bisacetamide, sodium butyrate, and cyclic AMP.
BACKGROUND
Hexamethylene bisacetamide (HMBA), sodium butyrate (NaBt), and cyclic AMP (cAMP) have been shown to induce differentiation, which may regulate tumour growth differently from conventional cytotoxic drugs. It was the intention in the present study to determine whether alterations could be induced in the phenotype of small cell lung cancer (SCLC) cell lines with HMBA, NaBt and cAMP, and whether these alterations would correlate with reduced growth in vivo, implying a phenotypic shift from malignancy towards differentiation.


METHODS
The cell lines were NCI-H69, H187 and H128. The activity of dopa decarboxylase (DDC), the BB isozyme of creatine kinase (CK-BB), the synthesis of bombesin-like peptide (BLI), and the presence of neurone specific enolase (NSE) and chromogranin were used as markers of the small cell phenotype. Clonogenicity in suspension in agar, and growth as xenografts in nude mice, were used as malignancy-associated properties. Cell proliferation in vitro was determined by cell counting and growth curve analysis.


RESULTS
HMBA, NaBt and cAMP were found to be reversibly cytostatic in liquid culture and pre-exposure reduced the cloning efficiency in agar by 60%-80%. Growth as xenografts was inhibited (three- to five-fold increase in the tumour doubling time), most significantly by NaBt. Effects of phenotypic markers were more complex. The most significant were a two-fold reduction in DDC with NaBt and HMBA, a 50% increase in CK-BB with cAMP, and a 70%-100% increase in secreted BLI with HMBA and cAMP, in NCI-H69 cells. No significant effects were seen on NSE and chromogranin. There was little sign of an interaction with adriamycin and vincristine, although a slight increase was observed in the ID50 of VP-16 following treatment with cAMP.


CONCLUSIONS
NaBt, HMBA and cAMP were cytostatic and inhibited tumour growth, but there was no coordinated response in marker expression that would confirm phenotypic alteration indicative of differentiation. The problem of defining differentiation in SCLC further complicated the analysis. The possibility remains of combining these agents with conventional cytotoxics as there appears to be little antagonistic effect, and other studies have suggested synergism may be possible with correct scheduling.
CRC Department of Medical Oncology, University of Glasgow, U.K.
M Z
MZ
Khan
R I
RI
Freshney
A M
AM
McNicol
A M
AM
Murray
D000081:Acetamides / Q000494:pharmacology*; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D014408:Biomarkers, Tumor / Q000378:metabolism; D002087:Butyrates / Q000494:pharmacology*; D020148:Butyric Acid; D018288:Carcinoma, Small Cell / Q000378:metabolism / Q000473:pathology*; D002454:Cell Differentiation / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D000242:Cyclic AMP / Q000494:pharmacology*; D004347:Drug Interactions; D006801:Humans; D008175:Lung Neoplasms / Q000378:metabolism / Q000473:pathology*; D051379:Mice; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D010641:Phenotype; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D014410:Tumor Stem Cell Assay
D000081:Acetamides; D000970:Antineoplastic Agents; D014408:Biomarkers, Tumor; D002087:Butyrates; D020148:Butyric Acid; D000242:Cyclic AMP; C014026:hexamethylene bisacetamide