14-3-3β Promotes Migration and Invasion of Human Hepatocellular Carcinoma Cells by Modulating Expression of MMP2 and MMP9 through PI3K/Akt/NF-κB Pathway

<div><p>14-3-3β has been demonstrated to possess the oncogenic potential, and its increased expression has been detected in multiple types of carcinomas. However, majority of previous studies focused on the role of 14-3-3β in tumor cell proliferation and apoptosis, leaving much to be elucidated about its function in tumor cell invasion and metastasis. Hence, the present study aimed to investigate the role of 14-3-3β in the invasion of hepatocellular carcinoma (HCC) cells and the implications in the prognosis of HCC patients. We first examined the expression of 14-3-3β in the primary tumors of HCC patients with or without portal vein tumor thrombus (PVTT), and found that 14-3-3β expression was higher in the primary tumors with PVTT, and the level was even higher in the PVTTs. Kaplan-Meier curves and multivariate analysis revealed that high expression of 14-3-3β was associated with overall survival (OS) and time to recurrence (TTR) of HCC patients. In addition, ectopic expression of 14-3-3β in HCC cell lines led to enhanced migration ability and invasiveness, as well as up-regulation of matrix metalloproteinase 2 and 9, which could be suppressed by inhibiting the activation of Akt and nuclear factor-κB (NF-κB) signaling. Furthermore, we identified a correlated elevation of 14-3-3β and p-Akt in the primary tumors of HCC patients, and showed that a combinatory detection of 14-3-3β and p-Akt could better predict post-surgical outcome of HCC patients.</p></div

Correlated elevation of 14-3-3β and p-Akt in primary HCC tumors predicts a worse post-surgical outcome in HCC patients.

<p><b>(A)</b> The levels of 14-3-3β and p-Akt in the primary HCC tumors were examined by tissue microarray. <b>(B)</b> The correlation of the levels between 14-3-3β and p-Akt in HCC tumors was analyzed using the Spearman correlation test. Kaplan-Meier curves of <b>(C)</b> cumulative post-surgical survival and <b>(D)</b> cumulative post-surgical recurrence in HCC patients grouped by the high or low levels of 14-3-3β and p-Akt.</p

14-3-3β-regulated expression of MMP2 and MMP9 is dependent on PI3K/Akt/NF-κB signaling pathway.

<p><b>(A)</b> Expression of 14-3-3β, MMP2 and MMP9 in 14-3-3β-transfected SMCC-7721 and Hep3B cells. <b>(B)</b> Western blot analysis of phosphorylation status of various signaling proteins and the level of nuclear p65 in 14-3-3β-overexpressing HCC cells. β-actin was used as the internal control for total or cytosolic proteins, while Lamin A served as the internal control for nuclear proteins. The 14-3-3β-transfected Hep3B cells were treated with <b>(C)</b> LY294002 (20 μM), the inhibitor of PI3K/Akt signaling, or <b>(D)</b> PDTC (10 μM), the inhibitor of NF-κB signaling, for 5 h, followed by western blot analysis of the expression of MMP2 and MMP9. <b>(E)</b> The invasiveness of 14-3-3β-overexpressing cells with and without LY294002 (20 μM) or PDTC (10 μM) treatment was assessed by Matrigel-based Transwell assay. Values are expressed as the mean ± standard deviation of three independent experiments. ***<i>p</i><0.001. <b>(F)</b> Western blot analysis was performed to determine the activation status of p-Akt and NF-κB in 14-3-3β-overexpressing cells after treating the cells with LY294002 (20 μM) for 5 h.</p

14-3-3β promotes migration and invasion of HCC cells.

<p><b>(A)</b> Matrigel-free Transwell assay was conducted to assess the migration ability of SMCC-7721 and Hep3B cells that were transfected with 14-3-3β expression construct or the empty vector pcDNA3.1. (B) Matrigel-based Transwell assay was performed to examine the invasiveness of SMCC-7721 and Hep3B with exogenous expression of 14-3-3β. <b>(C)</b> Matrigel-free migration assay and <b>(D)</b> Matrigel-based invasion assay were performed in CSQT-2 cells with and without expression of 14-3-3β-specific siRNA. This figure shows the representative images and the statistical analysis of three independent experiments. Values are expressed as the mean ± standard deviation. ***<i>p</i><0.001.</p

Increased expression of 14-3-3β in primary HCC tumors, PVTTs and HCC cell lines.

<p>(<b>A)</b> Real-time PCR and <b>(B)</b> western blot analysis of mRNA and protein levels of 14-3-3β in primary tumors (T), matched adjacent nontumorous liver tissues (N) and PVTTs (P) in HCC patients. (<b>C)</b> Western blot analysis of 14-3-3β expression in several HCC cell lines. β-actin was used as the internal control for all analysis. *<i>p</i><0.05; **<i>p</i><0.01.</p

High expression of 14-3-3β in primary HCC tumors is associated with poor survival and high recurrence of HCC patients after surgical resection.

<p><b>(A)</b> Relative expression levels of 14-3-3β in the primary HCC tumors grouped by the presence or absence of different clinical parameters, with the means and 95% confidence intervals. *<i>p</i><0.05. <b>(B)</b> Immunohistochemical examination of 14-3-3β in primary HCC tumors. H, high; L, low. <b>(C, D)</b> Kaplan-Meier curves of cumulative post-surgical survival and recurrence in HCC patients with high or low level of 14-3-3β. <b>(E, F)</b> Multivariate analysis of parameters associated with the unfavorable post-surgical survival and recurrence. HR, hazard ratio; CI, confidence interval.</p

Capsule Shedding and Membrane Binding Enhanced Photodynamic Killing of Gram-Negative Bacteria by a Unimolecular Conjugated Polyelectrolyte

The development of new antimicrobial agents to treat infections caused by Gram-negative bacteria is of paramount importance due to increased antibiotic resistance worldwide. Herein, we show that a water-soluble porphyrin-cored hyperbranched conjugated polyelectrolyte (PorHP) exhibits high photodynamic bactericidal activity against the Gram-negative bacteria tested, including a multidrug-resistant (MDR) pathogen, while demonstrating low cytotoxicity toward mammalian cells. Comprehensive analyses reveal that the antimicrobial activity of PorHP proceeds via a multimodal mechanism by effective bacterial capsule shedding, strong bacterial outer membrane binding, and singlet oxygen generation. Through this multimodal antimicrobial mechanism, PorHP displays significant performance for Gram-negative bacteria with >99.9% photodynamic killing efficacy. Overall, PorHP shows great potential as an antimicrobial agent in fighting the growing threat of Gram-negative bacteria

In Situ Noninvasive Observation of Nitric Oxide Fluctuation in SARS-CoV‑2 Infection In Vivo by Organic Near-Infrared-II Fluorescent Molecular Nanoprobes

The pathogenesis understanding of SARS-CoV-2 infection is crucial to prevent the rampant spread of COVID-19 and its contribution to deterioration in health, even death. Nitric oxide (NO), a crucial molecule involved in signal transduction and cytotoxicity, is a possible key regulator in the occurrence and development of COVID-19. However, understanding the pathogenesis of NO in SARS-CoV-2 infection is still in its infancy due to the lack of suitable in situ monitoring probes of NO fluctuation in the complex SARS-CoV-2 infection environment in deep lung tissues. Herein, we developed an activatable near-infrared-II fluorescent molecular nanoprobe (OSNP) that uncages high-resolution and deep-tissue-penetrating near-infrared-II fluorescence signal in specific response to NO for in situ and noninvasive visualization of NO fluctuation in a SARS-CoV-2 infection mouse model in lung tissues. In vivo visualization revealed that the NO level is a positive relationship with SARS-CoV-2 infection progress. With the assistance of immuno-histochemical analyses, we uncovered the NO-involved pathological mechanism, that being the improved NO level is associated with an increase in inducible NO synthase rather than endothelial NO synthase. Our study not only provides the example of a near-infrared-II fluorescent imaging of NO in SARS-CoV-2 infection but also provides opportunities to uncover tunderlying pathomechanism of NO for SARS-Cov-2 infections

Additional file 1 of KHDRBS3 accelerates glycolysis and promotes malignancy of hepatocellular carcinoma via upregulating 14-3-3ζ

Supplementary Material

Characteristics of COPD Patients and Healthy Controls^a.

a<p>FEV₁, forced expiratory volume in 1 s; FVC, functional vital capacity. Date for FEV_1% and FVC % are expressed as mean ± SD.</p