Attenuated LKB1-SIK1 signaling promotes epithelial-mesenchymal transition and radioresistance of non-small cell lung cancer cells.

BACKGROUND: Radiotherapy is one of the main therapeutic approaches for non-small cell lung cancer (NSCLC). However, radioresistant cancer cells can eventually cause tumor relapse and even fatal metastasis. It is thought that radioresistance and metastasis could be potentially linked by epithelial-mesenchymal transition (EMT). In this study, we established radioresistant NSCLC cells to investigate the potential relationship among radioresistance, EMT, and enhanced metastatic potential and the underlying mechanism involving liver kinase B1 (LKB1)-Salt-inducible kinase 1 (SIK1) signaling. METHODS: The radioresistant cell lines A549R and H1299R were generated by dose-gradient irradiation of the parental A549 and H1299 cells. The radioresistance/sensitivity was evaluated by Cell Counting Kit-8 assay, apoptosis analysis, and/or clonogenic cell survival assay. The EMT phenotype and the signaling change were assessed by Western blotting. The abilities of invasion and migration were evaluated by transwell assays and wound healing assays. RESULTS: The radioresistant cell lines A549R and H1299R displayed mesenchymal features with enhanced invasion and migration. Mechanistically, A549R and H1299R cells had attenuated LKB1-SIK1 signaling, which leaded to the up-regulation of Zinc-finger E-box-binding homeobox factor 1 (ZEB1)--a transcription factor that drives EMT. Re-expression of LKB1 in A549R cells reversed the EMT phenotype, whereas knockdown of LKB1 in H1299R cells further promoted the EMT phenotype. Moreover, re-expression of LKB1 in A549 cells increased the radiosensitivity, whereas knockdown of LKB1 in H1299 cells decreased the radiosensitivity. CONCLUSIONS: Our findings suggest that attenuated LKB1-SIK1 signaling promotes EMT and radioresistance of NSCLC cells, which subsequently contributes to the enhanced metastatic potential. Targeting the LKB1-SIK1-ZEB1 pathway to suppress EMT might provide therapeutic benefits.

[Synthesis and screening of polyethylenimine as a carrier for gene transfer into cultured human tumor cells].

BACKGROUND & OBJECTIVE: Choosing suitable gene carrier is very important in gene therapy. Recently, polyethylenimine (PEI), a new polycation compound, is particularly attractive due to its high transduction efficiency and low toxicity. This study was to synthesize a series of PEI nanogels of different particle diameters by photochemistry, investigate the relation between the transfection efficiency and particle diameter, and screen ideal gene carrier. METHODS: PEI nanogels were synthesized by photochemistry. The particle diameter was detected by photo correlation spectroscopy (PCS). The spherical morphology of the nanogels was characterized by scanning electron microscopy (SEM), and confirmed by atomic force microscopy (AFM). Using PEI/DNA complex as a plasmid vector, the enhanced green fluorescence protein (EGFP) gene was transferred into Bel7402 and A549 cells. Gene expression was quantitatively evaluated by fluorescent microscopy and flow cytometry (FCM). RESULTS: The diameter of synthesized PEI nanogels was in the range of 80-200 nm, and most of them were globular. The delivery rate reached the maximum when using 4 microg PEI (86.9 nm) to transfer 2 microg EGFP: the delivery rates were (32.75+/-1.01)% for Bel7402 cells and (29.81+/-1.84)% for A549 cells when detected by fluorescent microscopy, and were (32.40+/-1.41)% for Bel7402 cells and (30.00+/-1.86)% for A549 cell when detected by FCM. There was no significant difference between PEI and LipofectamineTM 2000 in the transfection efficiency (P > 0.05). CONCLUSIONS: PEI nanogels synthesized by photochemistry are effective nonviral vectors for gene delivery into human tumor cells in vitro. The transfection efficiency of 86.9 nm PEI is the highest.