Gemcitabine Enhances Kras-MEK-Induced Matrix Metalloproteinase-10 Expression Via Histone Acetylation in Gemcitabine-Resistant Pancreatic Tumor-initiating Cells

Objectives: Advanced pancreatic ductal adenocarcinoma is resistant to systemic chemotherapy, resulting in a poor prognosis. We previously isolated a human pancreatic tumor-initiating cell line, KMC07, from a patient with acquired resistance to gemcitabine chemotherapy. To improve the anticancer effects of gemcitabine, we investigated the molecular mechanism of KMC07 cells' resistance to gemcitabine. Methods: KMC07 cellswere treatedwith gemcitabine, then gene expression and functional analyses performed using microarray, the quantitative polymerase chain reaction, immunoblotting, immunohistochemistry, chromatin immunoprecipitation, and cell transplantation into nude mice. Results: KMC07 cells, but not BxPC-3, PANC-1, MIA PaCa-2, orAsPC-1 cells, expressed matrix metalloproteinase-10 mRNA, the expression level of which was enhanced by gemcitabine. KMC07 cellswere shown to carry a constitutively active Kras mutation, and a MEK inhibitor suppressed matrix metalloproteinase-10 mRNA expression. Gemcitabine enhanced histone H3 acetylation at the matrix metalloproteinase-10 promoter, and a histone acetyltransferase inhibitor reduced gemcitabine-enhanced matrix metalloproteinase-10 mRNA expression. Gemcitabine induced expression of matrix metalloproteinase-10 protein in KMC07-derived pancreatic tumors in vivo. Conclusions: We demonstrated constitutive activation of the Kras-MEK matrix metalloproteinase-10 signaling pathway in KMC07 cells that was enhanced by gemcitabine through histone acetylation. Our resultsmay provide novel insights into gemcitabine-based treatment for gemcitabineresistant pancreatic ductal adenocarcinoma.