Role of myofibroblasts in innate chemoresistance of pancreatic carcinoma--epigenetic downregulation of caspases.
We recently reported on continuous tumor-stroma interactions essentially contributing to chemoresistance of pancreatic ductal adenocarcinoma (PDAC) cells. As demonstrated here, long-term coculture with pancreatic myofibroblasts representing the main stromal compartment of PDAC resulted in a chemoresistant phenotype in the pancreatic ductal epithelial cell line H6c7 as well as in the chemosensitive PDAC cell line T3M4. This involved a reduced expression of caspases and the caspase inducing transcription factor STAT1, both caused by diminished gene transcription. The DNA-methylation inhibitor 5-azadeoxycytidine enhanced caspase and STAT1 expression in cocultured H6c7 and T3M4 cells along with an increased chemosensitivity, indicating a role for CpG DNA-hypermethylation in the downregulation of these crucial apoptosis mediators. Cocultured H6c7 and T3M4 cells exhibited elevated nuclear levels of DNA-methyltransferase-1 (DNMT1). Silencing of DNMT1 expression by siRNA increased expression of caspases and STAT1 and restored chemosensitivity. In SCID mice, tumors arising from coinoculated T3M4 cells and myofibroblasts (co-tumors) responded less towards chemotherapy than mono-tumors, exhibiting decreased apoptosis, no remission and reduced expression of caspases and STAT1. These data underscore the role of myofibroblasts in chemoresistance of PDAC and point to the importance of caspases as central target structures of epigenetic regulation in this scenario. Furthermore, an activated microenvironment might apparently promote the manifestation of chemoresistance already in premalignant precursor cells at early stages of PDAC tumorigenesis.
Müerköster
Susanne Sebens
SS
Clinic of General Internal Medicine, Laboratory of Molecular Gastroenterology and Hepatology, UKSH-Campus Kiel, Kiel, Germany.
Werbing
Veronika
V
Koch
Dorothee
D
Sipos
Bence
B
Ammerpohl
Ole
O
Kalthoff
Holger
H
Tsao
Ming-Sound
MS
Fölsch
Ulrich R
UR
Schäfer
Heiner
H
D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D001374:Azacitidine / Q000031:analogs & derivatives / Q000494:pharmacology; D021441:Carcinoma, Pancreatic Ductal / Q000188:drug therapy / Q000201:enzymology / Q000235:genetics / Q000473:pathology; D020169:Caspases / Q000096:biosynthesis* / Q000235:genetics; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000201:enzymology; D018920:Coculture Techniques; D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000096:biosynthesis / Q000378:metabolism; D000077209:Decitabine; D003841:Deoxycytidine / Q000031:analogs & derivatives / Q000494:pharmacology; D015536:Down-Regulation; D019008:Drug Resistance, Neoplasm; D044127:Epigenesis, Genetic; D005047:Etoposide / Q000494:pharmacology; D005260:Female; D005347:Fibroblasts / Q000201:enzymology / Q000473:pathology*; D006801:Humans; D051379:Mice; D016513:Mice, SCID; D010190:Pancreatic Neoplasms / Q000188:drug therapy* / Q000201:enzymology / Q000235:genetics* / Q000473:pathology; D050794:STAT1 Transcription Factor / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection; D000093542:Gemcitabine
D050794:STAT1 Transcription Factor; D003841:Deoxycytidine; D005047:Etoposide; D000077209:Decitabine; D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases; C000619380:DNMT1 protein, human; C000619381:Dnmt1 protein, mouse; D020169:Caspases; D001374:Azacitidine; D000093542:Gemcitabine