<p>iNOS is associated with tumorigenicity as an independent prognosticator in human intrahepatic cholangiocarcinoma</p>

Abstract

BackgroundInducible nitric oxide synthase (iNOS) has supposed to implicate in inflammation, infection, liver cirrhosis, and neoplastic diseases. This study was designed to explore the biological and clinical function of iNOS in intrahepatic cholangiocarcinoma (ICC).MethodsRT-PCR (Real-time quantitative PCR) and immunohistochemical staining were used to analyze the expression of iNOS in ICC and adjacent tissues. CCK8, transwell assays, flow cytometry were conducted to detect the proliferation, apoptosis, cell cycle. Western blotting was performed to detect the expression of target proteins. Multivariate analyses were conducted to analysis associates between clinicopathological values and survival.ResultsWe found that levels of iNOS mRNA and protein were dramatically increased in ICC samples and positively correlated with complicated bile duct stone, differentiation, pathology T, pathology M, Wip1, MMP-2, and MMP-9. iNOS expression was significantly correlated with the poor survival of ICC patients. Furthermore, iNOS was high expression in ICC cell lines (QBC-939, ICC-9810, SSP-25) compare with human normal biliary epithelium cell line (HIBEpic); both iNOS knockdown and iNOS inhibitor (1400 W) suppressed cell proliferation, invasion, and migration though nitric oxide production in ICC cells. Down-regulation of iNOS also induced G0/G1 cell cycle arrest and ICC cell apoptosis. Moreover, iNOS knockdown treatment significantly decreased Wip1, MMP-9, and MMP-2 gene expression.ConclusionLowly expressed iNOS-inhibited proliferation yet promoted apoptosis of ICC cells. Our data show targeted inhibition of iNOS in ICC may have therapeutic value.

The Impact of Matrix Metalloproteinase-9 on the Sequential Steps of the Metastatic Process

“…In addition, HIF-1 triggers the production of nitric oxide, a gaseous mediator capable of activating latent MMP-9 by disrupting the bond between the catalytic zinc and the propeptide domain of the enzyme ( Figure 2 ) [ 63 ]. Of interest, MMP-9 and nitric oxide synthases frequently colocalize at the leading edge of invading tumor cells [ 64 ].…”

MicroRNA-129-2-3p directly targets Wip1 to suppress the proliferation and invasion of intrahepatic cholangiocarcinoma

“…Our previous findings indicate that Wip1 is involved in the tumorigenicity and invasion of human ICC at least in part through the MMP-2 signaling pathway [17]. Our further data show that both iNOS and Wip1 promote ICC cell migration and invasion by up-regulating MMP expression [18]. To evaluate the underlying mechanism, we find miR-129-2-3p directly targets wip1 to suppress the proliferation and invasion of intrahepatic cholangiocarcinoma.…”

“…Others and our previous studies showed that Wip1 were over expression in a wide range of tumor tissues including kidney [12], breast [13], lung [14], ovarian [15], neuroblastoma [16]. Our previous research showed that Wip1 was highly expressed in ICC tissues and cell lines and may be a key regulator in the tumorigenicity and invasion of human ICC [17,18]. However, the potential mechanism is unknown.…”

Adenosine Suppresses Cholangiocarcinoma Cell Growth and Invasion in Equilibrative Nucleoside Transporters-Dependent Pathway

“…We further investigated the underlying mechanism that makes most CCA cell lines sensitive to adenosine; a few CCA cell lines and imCho cell lines were resistant. The levels of proinflammatory molecules were investigated because inflammation was shown to be involved in CCA progression and aggressiveness [38][39][40]. Levels of nuclear factor-κB (NF-κB), cyclooxygenase 2 (COX2), inducible nitric oxide synthase (iNOS) and Notch1 were examined in five CCA cell lines and one imCho cell lines (Figure 5a,b).…”