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Adenocarcinoma (AD) and squamous cell carcinoma (SCC) are both classified as major forms of non-small cell lung cancer, but differences in clinical prognoses and molecular mechanisms are remarkable. Recent studies have supported the importance of understanding immune status in that it influences clinical outcomes of cancer, and immunotherapies based on the theory of “immune editing” have had notable clinical success. Our study aimed to identify specific long non-coding (lnc) RNAs that control key immune-related genes and to use them to construct risk models for AD and SCC. Risk scores were used to separate patients into high- and low-risk groups, and we validated the prognostic significance of both risk scores with our own cohorts. A Gene Set Enrichment Analysis suggested that the immune responses of patients in the AD high-risk group and the SCC low-risk group tended to be weakened. Evaluation of immune infiltration revealed that the degree of infiltration of dendritic cells is of particular importance in AD. In addition, prediction of responses to immune checkpoint inhibitor (ICI) treatments, based on the T Cell Immune Dysfunction and Exclusion and immunophenoscore models, indicated that deterioration of the immune microenvironment is due mainly to T cell exclusion in AD patients and T cell dysfunction in SCC patients and that high-risk patients with SCC might benefit from ICI treatment. The prediction of downstream targets via The Cancer Proteome Atlas and RNA-seq analyses of a transfected lung cancer cell line indicated that the lncRNA LINC00996 is a potential therapeutic target in AD.

Background: Lung cancer remains the most fatal tumorous disease in the worldwide. Among that, lung adenocarcinoma (LUAD) was the most common histological type. A precise and concise prognostic model was urgently needed of LUAD. We developed a 23-gene signature for prognosis prediction based on EMT, immune and stromal datasets.Methods: Univariate Cox regression analysis was performed to select genes which were significantly associated with overall survival (OS) of the TCGA LUAD cohorts. LASSO regression and multivariate Cox regression analysis was used to build the multi-gene signature. Enrichment analyses and a protein-protein interactions (PPI) network were performed to show the interaction and functions of the signature. A nomogram was developed based on risk score and other clinical features. Predictive performance of the signature was externally validated in two independent datasets from Gene Expression Omnibus (GSE37745 and GSE13213).Results: A total of 1334 EMT, immune and stromal associated genes were obtained. After LASSO regression and multivariate Cox regression analysis, a 23-gene signature for risk stratification was built. K-M curves showed that the patients with high risk had a poorer outcome. Finally, a nomogram was built to predict prognosis. The predictive performance of the 23-gene signature was confirmed in internal and external validation.Conclusion: We developed and verified a 23-gene signature based on EMT, immune and stromal gene sets. It provided a convenient and concise tool for risk stratificationand individual medicine.

Background: Adenocarcinoma (AD) accounts for more than 30% of pulmonary carcinoma with the highest morbidity and mortality in the world. LncRNA regulates various biological processes and shows great potential to become a therapeutic target for AD patients. Methods: In this research, we combined normal lung tissue sample from the Genotype-Tissue Expression (GTEx) database and AD cohort in the Cancer Genome Atlas (TCGA) project to screen the significant Long non-coding RNAs (lncRNA). Besides, Gene Expression Omnibus (GEO) database and in vitro experiment were applied to subsequent verification. Findings: After screening, we selected PCPB1-AS1 as the core of follow-up research. Bioinformatic analyses indicated that PCBP1-AS1 specifically influenced intratumoral infiltration of T regulatory cell (Treg), which was verified by the experiments based on our cohort and AD cell lines. Meanwhile, ceRNA network suggested that RORA might be potential target of PCBP1-AS1 to regulate the Treg infiltration. In addition, the prediction of response to immune checkpoint inhibitors (ICI) treatment indicated PCBP1-AS1 expression level was able to become reference for ICIs treatment application. Interpretation: The compelling evidence demonstrated he rich biological functions of PCBP1-AS1 and its strong possibility to become the potential target for AD therapy. Funding Statement: National Natural Science Foundation (81672288), Focus on Research and Development Plan in Shandong Province (2018GSF118028) and Natural Science Foundation of Shandong Province (ZR2019PH002). Declaration of Interests: The authors declare that they have no conflict of interest. Ethics Approval Statement: This research was approved by Ethics Review Committee of Shandong Provincial Hospital, and was performed based on the Declaration of Helsinki and Good Clinical Practice guidelines, as defined by the International Conference on Harmonisation.

NIN/RPN12 binding protein 1 (NOB1p) encoded by NOB1 has been found to be an essential factor in 26S proteasome biogenesis which participates in protein degradation. However, the functions of NOB1 in non-small cell lung cancer cells are largely unknown. In the present study, lentivirus-mediated NOB1 shRNA transfection in two non-small cell lung cancer cell lines (A549 and H1299) was accomplished, as determined by fluorescence imaging. Downregulation of NOB1 expression was confirmed by real-time PCR and western blotting. NOB1 silencing resulted in a significant decline in the proliferation and colony formation capability of non-small cell lung cancer cells. Moreover, flow cytometry showed that A549 cells were arrested in the G0/G1 phase of the cell cycle after NOB1 suppression. Furthermore, depletion of NOB1 resulted in a significant decrease in CDK4 and cyclin D1 expression. These results suggest that NOB1 may act as an important regulator in non-small cell lung cancer growth and could be a therapeutic target of non‑small cell lung cancer.

Human serum apotransferrin (hTF) binds to Zr(IV) slowly in the presence of nitrilotriacetate (NTA), citrate or ethylenediaminetetraacetate (EDTA) as donor ligands. For Zr(NTA)(2)(2-) as donor, equilibrium was reached in ca. 2 h (pH 7.4, 298 K, 10 mM Hepes, 5 mM bicarbonate) and full loading of the N- and C-lobe sites was achievable to give Zr(2)-hTF. (13)C NMR data suggest that carbonate can bind as a synergistic anion. (1)H and 2D [(1)H,(13)C] (using epsilon-[(13)C]Met-hTF) NMR studies show that there is little lobe-selectively in the order of Zr(IV) uptake. Fe(III) displaced Zr(IV) from the C-lobe of Zr(2)-hTF first, followed by the N-lobe. However, in the presence of a large excess of NTA, Zr(IV) binds to the N-lobe of holo-hTF (Fe(2)-hTF) first followed by the C-lobe. The (1)H and (13)C NMR chemical shift changes for epsilon-[(13)CH(3)] of Met464, which is close to the C-lobe site, are quite distinct from those observed previously for Al(III), Fe(III), Ti(IV), Ga(III) and Bi(III) binding to hTF, suggesting that Zr(IV) binding may not induce lobe closure [as observed previously for Hf(IV)]. This may affect receptor recognition and play a role in the different biological behaviour of Zr(IV) compared to Ti(IV).

The ferric ion-binding protein A (FbpA), a member of transferrin superfamily, is a periplasmic iron transporter employed by many Gram-negative pathogens. Our experiments indicated copper(ii) could bind with Neisseria gonorrhoeae FbpA (NgFbpA), and the binding constant reached up to (8.7 ± 0.2) × 10(8) M(-1)via UV-vis titration. The crystal structure of recombinant Cu-NgFbpA at 2.1 Å revealed that the oxo-Cu2 clusters (dinuclear centres) assembled in the iron binding cleft and were bound to the two adjacent tyrosine residues (Y195 and Y196) of the protein, two Cu ions coordinated with two tyrosines, Y195 and Y196, respectively, which was different from the binding model of Fe ion with FbpA, in which Y195 and Y196 coordinated together with one Fe ion. While this was similar to the binding of Zr and Hf ion clusters, Y195 and Y196 coordinated with two metal ions and the μ-oxo-bridges linking the metal ions. Structural superimposition demonstrated that oxo-Cu2-NgFbpA still keeping an open conformation, similar to the apo-form of NgFbpA. The structure presented additional information towards an understanding of the function of FbpA, and provided a detailed binding model for FbpA protein with the possible metal ions in a biological system.

This paper will be the first to discuss the in vivo and in vitro properties of a Pd(II) complex, K2PdCl4, interacting with metallothioneins (MTs). In vivo experiments revealed that intraperitoneal injections of K2PdCl4 into rabbits led to the simultaneous synthesis of Pd–MT in the kidney and Zn7MT in the liver. The renal Pd–MT complex contains 3.6 ± 0.3 Pd, 2.1 ± 0.2 Zn, and 1.0 ± 0.1 Cu per mole protein. It was found that pre-treatment with Zn(NO3)2 before K2PdCl4 injections significantly enhanced renal Pd–MT level. The same pre-treatment also increases hepatic Zn–MT levels. These results strongly suggest that Pd(II) ions can be bound in vivo by MT existing in the rabbit kidneys to form Pd–MT. Gel-filtration chromatographic studies after the incubation of either native Cd5Zn2MT2 or Zn7MT2 with K2PdCl4 in vitro demonstrate that Pd(II) ions promote the non-oxidative oligomerization of native MTs. Increasing the level of Pd(II) relative to MT led to a concomitant increase in the apparent yield of MT oligomers. At relatively low Pd–MT ratio, Pd(II) is found predominantly in the oligomers while the monomeric products are chiefly composed of the reactants, Cd5Zn2MT2 or Zn7MT2. Based on our experimental data, the mechanisms of the reactions between Pd(II) and MTs in vivo and in vitro are discussed.