Insulin promotes invasion and migration of KRAS^G12D mutant HPNE cells by upregulating MMP‐2 gelatinolytic activity via ERK‐ and PI3K‐dependent signalling

Abstract

Objectives: Hyperinsulinemia is a risk factor for pancreatic cancer, but the function of insulin in carcinogenesis is unclear, so this study aimed to elucidate the carcinogenic effects of insulin and the synergistic effect with the KRAS mutation in the early stage of pancreatic cancer. Materials and methods:A pair of immortalized human pancreatic duct-derived cells, hTERT-HPNE E6/E7/st (HPNE) and its oncogenic KRAS G12D variant, hTERT-HPNE E6/ E7/KRAS G12D /st (HPNE-mut-KRAS), were used to investigate the effect of insulin. Cell proliferation, migration and invasion were assessed using Cell Counting Kit-8 and transwell assays, respectively. The expression of E-cadherin, N-cadherin, vimentin and matrix metalloproteinases (MMP-2, MMP-7 and MMP-9) was evaluated by Western blotting and/or qRT-PCR. The gelatinase activity of MMP-2 and MMP-9 in conditioned media was detected using gelatin zymography. The phosphorylation status of AKT, GSK3β, p38, JNK and ERK1/2 MAPK was determined by Western blotting. Results: The migration and invasion ability of HPNE cells was increased after the intro-duction of the mutated KRAS gene, together with an increased expression of MMP-2.These effects were further enhanced by the simultaneous administration of insulin.The use of MMP-2 siRNA confirmed that MMP-2 was involved in the regulation of cell invasion. Furthermore, there was a concentration-and time-dependent increase in gelatinase activity after insulin treatment, which could be reversed by an insulin receptor tyrosine kinase inhibitor (HNMPA-(AM) 3 ). In addition, insulin markedly enhanced the phosphorylation of PI3K/AKT, p38, JNK and ERK1/2 MAPK pathways, with wortmannin or LY294002 (a PI3K-specific inhibitor) and PD98059 (a MEK1-specific inhibitor) significantly inhibiting the insulin-induced increase in MMP-2 gelatinolytic activity.

lncRNA MIAT promotes cell invasion and migration in esophageal cancer

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Insulin negatively regulates dedifferentiation of mouse adipocytes in vitro

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The Role of Cell Proliferation and Extracellular Matrix Accumulation Induced by Food Additive Butylated Hydroxytoluene in Uterine Leiomyoma

“…PI3K inhibitor, wortmannin, was used to investigate the potential modulator of BHT on ECM accumulation. According to a previous study, PI3K acts as the important modulator of MMP-2 [ 27 ]. By using wortmannin, the results indicated that the PI3K inhibition could reverse the BHT’s effect on ECM accumulation ( Figure 5 ).…”