A Risk Score Based on Immune- and Oxidative Stress-Related LncRNAs Predicts Prognosis in Lung Adenocarcinoma: Insights from in vitro Experiments and Large-Scale Transcriptome Analysis.

Background: Long non-coding RNAs (lncRNAs) were demonstrated to be key to cancer progression and highly associated with the tumor immune microenvironment. Oxidative stress and immune may modulate the biological behaviors of tumors. Therefore, biomarkers that combined oxidative stress, immune, and lncRNA can be a promising candidate bioindicator in clinical therapy of cancers. Methods: Immune-related genes (IRGs) and oxidative stress-related genes (ORGs) were identified based on a detailed review of published literatures. The transcriptome data and clinical information of lung adenocarcinoma (LUAD) patients were obtained from TCGA database. Lasso and Cox regression analyses were conducted to develop a prognostic model. Additionally, the link between immune checkpoints, immune cells, and the prognostic model was investigated, and predict the sensitivity of immunotherapy. Results: 2498 IRGs and 809 ORGs were extracted from previous studies, and 190 immune- and oxidative stress-related genes (IOGs) were acquired by overlapping the above genes. 658 immune- and oxidative stress-related lncRNAs (IOLs) were screened by Pearson correlation analysis. A total of 25 prognosis-related IOLs were screened by univariate regression analysis. Finally, LASSO Cox regression analysis was adopted for determining a 12-IOLs prognostic risk signature. The signature performance was confirmed in the training cohort and the testing cohort, and cases were classified into low- and high-risk groups by the risk score calculated from the signature. Patients in the high-risk group had poor prognoses and immunosuppression, while the risk score was significantly associated with tumor-infiltrating immune cells, immune checkpoint expression, and immunotherapy responses. In vitro experiments further confirmed the expression of key signature gene. Conclusion: Our new IOLs-related prognostic signature can be reliable prognostic tools and therapeutic targets for LUAD patients.

Novel histone acetylation-related lncRNA signature for predicting prognosis and tumor microenvironment in esophageal carcinoma.

Histone acetylation is one of the most common epigenetic modifications and plays a crucial role in tumorigenesis. However, the prognostic significance of histone acetylation-related lncRNAs (HARlncRNAs) in esophageal carcinoma (ESCA) is not well understood. A total of 653 differentially expressed lncRNAs (DElncRNAs) were identified between 162 ESCA tissues and 11 normal tissues in the TCGA database, and 7 of them were correlated with acetylation regulators. We employed univariate Cox regression analysis, combining it with clinical prognosis information, to select 3 prognostic-related HARlncRNAs for further analysis. Subsequently, we used LASSO regression analysis to construct a risk signature for ESCA and identified C21orf62-AS1 and SSTR5.AS1 as potential biomarkers for the prognosis of ESCA patients. Based on the risk score calculated using the risk signature, we categorized patients into high- and low-risk groups. We identified the risk score as an independent risk factor and validated it in the training, test, and GSE53624 datasets. Additionally, patients categorized by their risk scores exhibited distinct immune statuses, tumor mutation burdens, responses to immunotherapy, and drug sensitivities.

The regulation of autophagy by the miR-199a-5p/p62 axis was a potential mechanism of small cell lung cancer cisplatin resistance.

BACKGROUND: Autophagy has been found to be involved in the multidrug resistance (MDR) of cancers, but whether it is associated with resistance of small cell lung cancer (SCLC) has not been studied. Here, we hypothesized that a potential autophagy-regulating miRNA, miR-199a-5p, regulated cisplatin-resistant SCLC. METHODS: We validated the MDR of H446/EP using CCK-8 and LDH. We tested the binding of miR-199a-5p to p62 using the Dual-Luciferase assay and validated the association of miR-199a-5p and p62 in SCLC samples. We overexpressed (OE) and knocked down (KD) miR-199a-5p in H446 and H446/EP and determined the expression of miR-199a-5p, autophagy-related proteins, and the formation of autophagolysosomes using QPCR, western blotting, and MDC staining respectively. These results were validated in an orthotopic H446 mouse model of SCLC. RESULTS: H446/EP was resistant to cisplatin, etoposide, paclitexal, epirubicin, irinotecan, and vinorelbine. Exposure of cisplatin at 5 µg/ml for 24 h increased LC3II/LC3I, ATG5, p62, and the formation of autophagolysosomes in H446 cells, but not in H446/EP cells. The expression of miR-199a-5p was up-regulated in H446/EP compared to H446. MiR-199a-5p directly targeted the p62 gene. The expression of miR-199a-5p and p62 were correlated in SCLC samples. In H446 and H69PR, the OE of miR-199a-5p increased LC3II/LC3I, p62, and the formation of autophagolysosomes, but not ATG5, while the KD of miR-199a-5p decreased p62, but did not affect LC3II/LC3I, ATG5, and the formation of autophagolysosomes. In H446/EP, the OE of miR-199a-5p decreased p62 only. These results were generally consistent to results in the animal tumor samples. CONCLUSIONS: The regulation of autophagy by the miR-199a-5p/p62 axis was a potential mechanism of small cell lung cancer cisplatin resistance.

miR-493 by regulating of c-Jun targets Wnt5a/PD-L1-inducing esophageal cancer cell development.

BACKGROUND: Esophageal cancer is one of the most common cancers across the globe; the 5-year survival of esophageal cancer patients is still low. MicroRNA (miRNA) dysregulation has been implicated in cancer development, and the miRNAs play a pivotal role in esophageal cancer pathogenesis. It is urgently needed to find out how miRNA dysregulation was involved in esophageal cancer (EC) development. METHODS: Through experiments in vivo and in vitro, we explored potential signaling pathways, miR-493/Wnt5A/c-JUN loop, in EC. Their mechanistic roles in EC cell proliferation, migration, and invasion were investigated through multiple validation steps in EC9706 and TE13 cell lines and EC specimens. RESULTS: Overexpression of miR-493 attenuates esophageal cancer cell proliferation, migration, and invasion in vivo and in vitro. Moreover, miR-493 downregulation is an unfavorable factor in EC and negatively correlated with Wnt5A. The existence of miR-493 is also an important attribute of metabolism. Based on mechanism analyses, we show that miR-493 inhibits the activity of c-JUN and p-PI3K/p-AKT with enhanced p21 and directly regulates Wnt5A expression and function, whereas c-JUN binds the promoter region of miR-493 and suppressed the expression of miR-493, forming a negative feedback loop. CONCLUSIONS: The miR-493/Wnt5A/c-JUN loop is a molecular feedback loop that refers to the development of esophageal cancer cells and a potential target for the treatment of esophageal cancer.

[Exploration of the Suitable Culture Conditions for Extracellular Microvesicles Derived from Human Mesenchymal Stem Cells].

OBJECTIVE: To explore the appropriate procedures for preparing extracellular microvesicles (MV) derived from human mesenchymal stem cells (MSC). METHODS: Human MSCs from umbilical cords were cultured in a serum-free medium and maintained in a basal medium for 72 hours after the cell confluence reached to 80%. The supernatants of cultured cells were collected and MVs were enriched. MVs were identified by flow cytometry and electron microscopy. The total protein amount in MVs was used as a parameter for the content of MVs. The supernatants were adjusted to different pH values, and the output of MVs was detected. The supernatants were also collected for enriching the MV and detecting the protein content of MV after the cells were maintained in the basic medium for different time. RESULTS: Flow cytometric analysis showed that the MVs expressed CD9, CD63 and CD81, morphologically presented round under an electron microscope and the diameter of MV was around 100 nm. After enrichment of MV, the protein content of MVs in the supernatants was 416.8±128.1, 255.4±77.9 and 142.8±46.4 µg per 107 MSC，respectively at pH of supernatant 3, 7 and 9 (P＜0.05). The protein content of the supernatants per 107 MSC was 173.6±44.5, 262.4±49.6 and 364.2±37.8 µg respectively after starvation culture for 48, 72 and 96 hrs (P＜0.05). CONCLUSION: MVs can be readily collected after MSCs were starved for 96 hours, and the pH of the supernatants is adjusted at 3.0.

Exploring the microRNA profiles as potential diagnostic probes for oligo- and polymetastatic prognosis of lung metastasis(es) patients.

Presuming the stage of metastatic lung cancer is divided by its location, an intermediate state of ≤5 cumulative metastasis is defined as oligometastases (OM) and a widespread state of >5 cumulative metastasis as polymetastases (PM). According to the phenotypes, the different metastatic cancer patients can be treated with different methods: the OM patients can be treated by a metastasis-directed local therapy method, whereas the PM patients are not recommended to take such a treatment. It is also believed that the patients at the initial OM stage may progress to the PM stage. Currently, the OM- and PM-metastatic cancer patients can be identified by traditional imaging methods. However, the current methods are found to be insufficient for the discrimination. It hence is meaningful and important to develop new diagnostic methods for a better prediction to the patients following by selecting a correct metastasis-directed treatment.MicroRNAs (miRNAs) can be used as the genetic probes for the new diagnostic methods. In this study, a bioinformatics strategy was employed to screen the microRNAs as potential diagnostic probes for distinguishing the OM and PM lung metastases patients. The expression profiles of microarray data of GSE38698 were downloaded from Gene Expression Omnibus (http://www.ncbi.nlm.nih.gov/geo/) including the information from 63 patients: 24 PM and 39 OM patients. The microRNA expression patterns of tumor samples were identified for the OM and PM patients who were treated with the high-dose radiotherapy. Followed by analyzing the functional enrichment pathways, an early diagnosis model of OM and PM groups was identified with different expression genes (DEGs). The ratios of PM/OM were calculated by setting a high significance in the expressions of 377 mature miRNAs in the profile [log2 (PM/OM) >1 and P < .05]. Through a high combination power [area under the curve (AUC) ≥ 0.875] with the superior sensitivity and specificity, a panel of 10 miRNAs including 7 upregulation and 3 downregulation expressions were identified as potential probes for discriminating the PM and OM patients from the receiving operation characteristic (ROC). Considering the possible involvements of cancer progress, the interconnected axon guidance, cancer metastasis pathways, proteoglycans, and Mitogen-activated protein kinases signaling pathway and endocytosis were suggested for the subsequent miRNA target analysis. The results may reveal a biological significance that a profile of miRNAs can be used as the potential probes to identify the patients at the OM or PM stages and figure out the metastasis-directed treatment methods for the patients at the different metastasis stages.

Nickel-Mediated Asymmetric Allylic Alkylation between Nitroallylic Acetates and Acyl Imidazoles.

A nickel-mediated asymmetric allylic alkylation reaction between imidazole-modified ketones and nitroallylic acetates is presented. This reaction is catalyzed by a simple chiral diamine-nickel catalyst under mild conditions and leads to a series of novel enantioenriched α-allylic adducts in moderate to good yields with excellent enantioselectivities. Furthermore, transformation of the allylic adducts could smoothly lead to chiral γ-nitro-esters containing three continuous stereocenters in good yields.

An Efficient Nickel-Catalyzed Asymmetric Oxazole-Forming Ugi-Type Reaction for the Synthesis of Chiral Aryl-Substituted THIQ Rings.

A nickel-catalyzed asymmetric oxazole-forming Ugi reaction of C,N-cyclic azomethine imines and isonitriles is disclosed. The reported protocol proceeds smoothly, and gives the corresponding adducts, which contain two important pharmaceutically active ring-systems (tetrahydroquinoline and oxazole rings), in good yields and excellent enantioselectivities by employing an easily accessible chiral diamine as a ligand. This simple and efficient strategy provides easy access to a series of C1-substituted aryl tetrahydroisoquinolines.

Potential sodium D2 resonance radiation generated by intra-cavity SHG of a c-cut Nd:YVO4 self-Raman laser.

Intra-cavity frequency doubling with 589 nm emission from a compact c-cut Nd:YVO4 crystal self-Raman laser was investigated. A 15-cm-length LBO with non-critical phase-matching cut (θ = 90°, ϕ = 0°) was used for efficient second-harmonic generation. At a pump power of 16.2 W and a pulse repetition frequency of 40 kHz, output power up to 2.15 W was achieved with a pulse width of 16 ns and a conversion efficiency of 13.3% with respect to the diode pump power. The center wavelength was measured to be 589.17 nm with a Half-Maximum-Full-Width of 0.2 nm, which was well in accordance with the sodium D2 resonance radiation.