title: Cancer Associated Fibroblast-Derived Hepatocyte Growth Factor Inhibits the Paclitaxel-Induced Apoptosis of Lung Cancer A549 Cells by Up-Regulating the PI3K/Akt and GRP78 Signaling on a Microfluidic Platform.
abstract: Tumor stroma and growth factors provide a survival environment to tumor cells and can modulate their chemoresistance by dysregulating several signal pathways. In this study, we fabricated a three-dimensional (3D) microfluidic chip using polydimethylsiloxane (PDMS) to investigate the impact of hepatocyte growth factor (HGF) from cancer-associated fibroblasts (CAF) on the Met/PI3K/AKT activation, glucose regulatory protein (GRP78) expression and the paclitaxel-induced A549 cell apoptosis. With a concentration gradient generator, the assembled chip was able to reconstruct a tumor microenvironment in vitro. We found high levels of HGF in the supernatants of CAF and the CAF matrix from the supernatants of activated HFL1 fibroblasts or HGF enhanced the levels of Met, PI3K and AKT phosphorylation and GRP78 expression in A549 cells cultured in a 3D cell chamber, which was abrogated by anti-HGF. Inhibition of Met attenuated the CAF matrix-enhanced PI3K/AKT phosphorylation and GRP78 expression while inhibition of PI3K reduced GRP78 expression, but not Met phosphorylation in A549 cells. Inhibition of GRP78 failed to modulate the CAF matrix-enhanced Met/PI3K/AKT phosphorylation in A549 cells. Furthermore, inhibition of PI3K or GRP78 enhanced spontaneous and paclitaxel-induced A549 cell apoptosis. Moreover, treatment with the CAF matrix inhibited spontaneous and medium or high dose of paclitaxel-induced A549 cell apoptosis. Inhibition of PI3K or GRP78 attenuated the CAF matrix-mediated inhibition on paclitaxel-induced A549 cell apoptosis. Our data indicated that HGF in the CAF matrix activated the Met/PI3K/AKT and up-regulated GRP78 expression, promoting chemoresistance to paclitaxel-mediated apoptosis in A549 cells. Our findings suggest that the microfluidic system may represent an ideal platform for signaling research and drug screening.
authors.affiliation: Department of Gastroenterology, the Second Hospital of Dalian Medical University, Dailan, China.
authors.forename: Li
authors.initials: L
authors.lastname: Ying
authors.affiliation: Department of Respiratory, the Second Hospital of Dalian Medical University, Dailan, China.
authors.forename: Ziwei
authors.initials: Z
authors.lastname: Zhu
authors.affiliation: Department of Respiratory, the Second Hospital of Dalian Medical University, Dailan, China.
authors.forename: Zhiyun
authors.initials: Z
authors.lastname: Xu
authors.affiliation: School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Tianrui
authors.initials: T
authors.lastname: He
authors.affiliation: Department of Respiratory, the Second Hospital of Dalian Medical University, Dailan, China.
authors.forename: Encheng
authors.initials: E
authors.lastname: Li
authors.affiliation: Department of Respiratory, the Second Hospital of Dalian Medical University, Dailan, China.
authors.forename: Zhe
authors.initials: Z
authors.lastname: Guo
authors.affiliation: Department of Respiratory, the Second Hospital of Dalian Medical University, Dailan, China.
authors.forename: Fen
authors.initials: F
authors.lastname: Liu
authors.affiliation: Department of Gastroenterology, the Second Hospital of Dalian Medical University, Dailan, China.
authors.forename: Chunmeng
authors.initials: C
authors.lastname: Jiang
authors.affiliation: Department of Respiratory, the Second Hospital of Dalian Medical University, Dailan, China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Wang
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D004195:Disease Models, Animal; D019008:Drug Resistance, Neoplasm; D000091342:Endoplasmic Reticulum Chaperone BiP; D004789:Enzyme Activation / Q000187:drug effects; D005346:Fibroblast Growth Factors / Q000378:metabolism* / Q000494:pharmacology; D005347:Fibroblasts / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000378:metabolism*; D006801:Humans; D044085:Microfluidics / Q000379:methods; D009369:Neoplasms / Q000378:metabolism* / Q000473:pathology; D017239:Paclitaxel / Q000494:pharmacology*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects*; D023041:Xenograft Model Antitumor Assays
chemical_list: D000091342:Endoplasmic Reticulum Chaperone BiP; D005346:Fibroblast Growth Factors; D006360:Heat-Shock Proteins; D017239:Paclitaxel; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt; C000717247:HSPA5 protein, human