Molecular Analysis of Prognosis and Immune Infiltration of Ovarian Cancer Based on Homeobox D Genes.

Background: Homeobox D (HOXD) genes were associated with cancer pathogenesis. However, the role of HOXD genes in ovarian cancer (OC) and the possible mechanisms involved are unclear. In this study, we analyzed the function and regulatory mechanisms and functions of HOXD genes in OC based on comprehensive bioinformatics analysis. Methods: Expression of HOXD1/3/4/8/9/10/11/12/13 mRNA was analyzed between OC tissue and normal tissue using ONCOMINE, GEO, and TCGA databases. The relationship between HOXD expression and clinical stage was studied by GEPIA. The Kaplan-Meier plotter was used to analyze prognosis. cBioPortal was used to analyze the mutation and coexpression of HOXDs. GO and KEGG analyses were performed by the DAVID software to predict the function of HOXD coexpression genes. Immune infiltration analysis was used to evaluate the relationship between the expression of HOXD genes and 24 immune infiltrating cells. Results: The expression of HOXD3/4/8/9/10/11 was significantly lower in OC tissues than in normal ovarian tissues, while the expression of HOXD1/12/13 was significantly higher in OC tissues. The expression of HOXD genes was associated with FIGO stage, primary therapy outcome, tumor status, anatomic neoplasm subdivision, and age. The expression levels of HOXD1/3/4/8/9/10 correlated with tumor stage. HOXD1/8/9 could be served as ideal biomarkers to distinguish OC from normal tissue. Low HOXD9 expression was associated with shorter overall survival (OS) (HR: 0.75; 95% CI: 0.58-0.98; P = 0.034) and progression-free survival (PFS) (HR: 0.69; 95% CI: 0.54-0.87; P = 0.002). The HOXD coexpression genes were associated with pathways including cell cycle, TGF-beta signaling pathway, cellular senescence, and Hippo signaling pathway. HOXD genes were significantly associated with immune infiltration. Conclusion: The expression of HOXD genes is associated with clinical characteristics. HOXD9 is a new biomarker of prognosis in OC, and HOXD1/4/8/9/10 may be potential therapeutic targets. The members of the HOXD genes may be the response to immunotherapy for OC.

TRIM21 deficiency promotes cell proliferation and tumorigenesis via regulating p21 expression in ovarian cancer.

Tripartite motif-containing 21 (TRIM21) has been reported to have a cancer-promoting or anticancer effect in various tumors; however, its role in ovarian cancer (OC) remains to be elucidated. In this study, we explored the biological function of TRIM21 in OC progression and investigated the potential mechanisms. We found that TRIM21 was remarkably decreased in OC tissues and cell lines compared with adjacent-cancerous tissues and normal ovarian epithelium cell. Decreased expression of TRIM21 in OC patients was significantly correlated with shorter overall and disease-specific survival by The Cancer Genome Atlas database (TCGA) analysis. Functional assays revealed that TRIM21 inhibited the migration and invasion of OC cells; and that TRIM21 also obviously impaired cell proliferation by inhibiting cell cycle progression in vitro and in vivo. Taken together, our results suggest that TRIM21 may be a promising biomarker and target for OC diagnosis and treatment.

NPD1 Enhances Autophagy and Reduces Hyperphosphorylated Tau and Amyloid-ß42 by Inhibiting GSK3ß Activation in N2a/APP695swe Cells.

BACKGROUND: The most prevalent kind of dementia, Alzheimer's disease (AD), is a neurodegenerative disease. Previous research has shown that glycogen synthase kinase-3ß (GSK-3ß) is involved in the etiology and progression of AD, including amyloid-ß (Aß), phosphorylated tau, and mitochondrial dysfunction. NPD1 has been shown to serve a neuroprotective function in AD, although the mechanism is unclear. OBJECTIVE: The effects of NPD1 on Aß expression levels, tau protein phosphorylation, apoptosis ratio, autophagy activity, and GSK-3ß activity in N2a/APP695swe cells (AD cell model) were studied, as well as the mechanism behind such effects. METHODS: N2a/APP695swe cells were treated with NPD1, SB216763, or wortmannin as an AD cell model. The associated proteins of hyperphosphorylated tau and autophagy, as well as the activation of GSK3ß, were detected using western blot and RT-PCR. Flow cytometry was utilized to analyze apoptosis and ELISA was employed to observe Aß42. Images of autophagy in cells are captured using transmission electron microscopy. RESULTS: In N2a/APP695swe cells, NPD1 decreased Aß42 and hyperphosphorylated tau while suppressing cell death. NPD1 also promoted autophagy while suppressing GSK-3ß activation in N2a/APP695swe cells. The outcome of inhibiting GSK-3ß is comparable to that of NPD1 therapy. However, after activating GSK-3ß, the opposite experimental results were achieved. CONCLUSION: NPD1 might minimize cell apoptosis, downregulate Aß expression, control tau hyperphosphorylation, and enhance autophagy activity in AD cell models to promote neuronal survival. NPD1's neuroprotective effects may be mediated via decreasing GSK-3ß.

Curcumin Decreases Hyperphosphorylation of Tau by Down-Regulating Caveolin-1/GSK-3ß in N2a/APP695swe Cells and APP/PS1 Double Transgenic Alzheimer's Disease Mice.

Caveolin-1, the marker protein of membranal caveolae, is not only involved in cholesterol regulation, but also participates in the cleavage of amyloid [Formula: see text]-protein precursor (APP) and the generation of [Formula: see text]-amyloid peptide. It has been reported to be tightly related with Tau. In our previous studies, curcumin has been confirmed to play a neuroprotective role in Alzheimer's disease (AD), but its effects on Caveolin-1, Tau and their correlation, and the mechanism is still unknown. As such, in the present study, N2a/WT cells, N2a/APP695swe cell and six-month-old APP/PS1 double transgenic mice were enrolled. After curcumin treatment, the expression of Caveolin-1, Tau and their relationship was detected, and the potential mechanisms were explored. The results showed that in the N2a/APP695swe cells, curcumin not only decreased the number of caveolae, but also made their membrane to be thinner; and curcumin could decreased the expression of phosphorylated Tau (P-Tau(ser404)/Tau) and Caveolin-1 ([Formula: see text]), but the expression of phosphorylated GSK-3[Formula: see text] (P-GSK-3[Formula: see text]/GSK-3[Formula: see text] was increased ([Formula: see text]). In APP/PS1 transgenic mice, the same results were observed. Taken together, our data suggest that curcumin may play an important role in AD via reducing Caveolin-1, inactivating GSK-3[Formula: see text] and inhibiting the abnormal excessive phosphorylation of Tau, which will provide a new theory for AD treatment with curcumin.

Lactate induces osteoblast differentiation by stabilization of HIF1α.

Aerobic glycolysis is involved in osteoblast differentiation induced by Wnt signaling or PTH treatment. However, it is still unclear whether lactate, the end product of aerobic glycolysis, plays any role in osteoblast differentiation. Herein we report that in cultures of osteoblast-lineage cells, lactate promoted alkaline phosphatase-positive cell formation, increased the activity of alkaline phosphatase, and induced the expression of osteocalcin. This osteoblast differentiation-inducing effect of lactate can be inhibited by blocking its entry into cells with MCT1 siRNA or inhibitors, and by interfering with its metabolism by using specific siRNAs for LDHB and PDH. Moreover, lactate stabilized HIF1α expression and inhibited HIF1α activity, with BAY87-2243 lowering the osteoblast differentiation-inducing effect of lactate. Thus, these findings reveal an unrecognized role for aerobic glycolysis in osteoblast differentiation via its end product, lactate.

[Biliverdin protects the isolated lungs from ischemia/reperfusion injury via anti-apoptosis].

OBJECTIVE: To observe the effect of biliverdin (BV) on the lung ischemia/reperfusion injury (LIRI), and to investigate the mechanism of BV in treatment of LIRI. METHODS: Thirty-two male Sprague-Dawley (SD) rats were randomly divided into control group, LIRI group, glutathione (GSH) group and BV-treated group, with 8 rats in each group. The rat LIRI model was reproduced by isolated lung perfusion system. Fifteen minutes after perfusion balance, lungs of control group were perfused continuously for 90 minutes, and the lungs in other groups were reperfused for 90 minutes after 1-hour ischemia, while the perfusion was added 10 µmol/L BV in BV-treated group and 4 mmol/L GSH in GSH group respectively. During the perfusion, the respiratory related indicators, such as tidal volume (VT), static compliance (Cst), arterial partial pressure of oxygen (PaO2), airway resistance (Raw), were dynamically observed. After perfusion, the wet/dry weight ratio (W/D) of lungs was determined. Pathological changes in lung tissue were observed under light microscope. The cell apoptosis was observed by TdT-mediated dUTP nick end labeling (TUNEL) method, and the apoptosis index was calculated. The protein expression levels of heme oxygenase-1 (HO-1), phosphorylated c-Jun N-terminal kinase (p-JNK), and caspase-3 were determined by Western Blot. RESULTS: Compared with control group, VT, Cst and PaO2 in LIRI group were significantly decreased from reperfusion for 30 minutes, and Raw was significantly increased. The pathological results showed that there was different degree of hyperemia edema, inflammatory cells infiltration and bronchial endothelium injury in the lung tissue of LIRI group. The W/D ratio of LIRI group was significantly higher than that of control group (8.98±2.34 vs. 5.89±0.52, P < 0.05). TUNEL results showed that the tan apoptosis cells of LIRI group were more than control group, and the apoptosis index was significantly higher than that of the control group [(13.88±2.35)% vs. (2.26±0.60)%, P < 0.05]. Compared with control group, the protein expression levels of HO-1, p-JNK, and caspase-3 in LIRI group were significantly increased [HO-1 (gray value): 0.55±0.13 vs. 0.16±0.02, p-JNK (gray value): 0.46±0.08 vs. 0.16±0.05, caspase-3 (gray value): 0.65±0.13 vs. 0.26±0.03, all P < 0.05]. Compared with LIRI group, the respiratory indicators in BV-treated group were improved significantly, the lung tissue injury was significantly reduced, the W/D ratio was significantly decreased (6.39±0.45 vs. 8.98±2.34, P < 0.05), and the cell apoptosis index was significantly reduced [(4.49±1.10)% vs. (13.88±2.35)%, P < 0.05], as well as the protein expression levels of HO-1, p-JNK, and caspase-3 were significantly lowered [HO-1 (gray value): 0.19±0.03 vs. 0.55±0.13, p-JNK (gray value): 0.31±0.06 vs. 0.46±0.08, caspase-3 (gray value): 0.33±0.05 vs. 0.65±0.13, all P < 0.05], which indicating that BV could alleviate LIRI via anti-apoptosis. The improvement effect of BV on PaO2, apoptosis index, protein expressions of HO-1 and caspase-3, and JNK phosphorylation were better than positive drug GSH, indicating that BV could protect LIRI ideally. CONCLUSIONS: BV alleviates LIRI via its anti-JNK pathway and its anti-apoptosis property.

Neuroglobin Attenuates Beta Amyloid-Induced Apoptosis Through Inhibiting Caspases Activity by Activating PI3K/Akt Signaling Pathway.

Excessive accumulation and deposition of amyloid-beta (Aß) has been considered as a pivotal event in the pathogenesis of Alzheimer's disease (AD). Neuronal apoptosis is one of the characteristics of AD, which is a possible mechanism underlying Aß-induced neuronal neurotoxicity. Neuroglobin (Ngb) is a newly discovered vertebrate heme protein that exhibits neuroprotective functions against cell death associated with hypoxic and amyloid insult. However, until now, the exact mechanism of neuroglobin's protective action has not been determined. To investigate the potential neuroprotective roles and mechanisms of Ngb, transgenic AD mice (APPswe/PSEN1dE9) and SH-SY5Y cells transfected with pAPPswe were enrolled into the study. In vivo, overexpression of Ngb via intracerebroventricular injection with pNgb attenuated memory, cognitive impairment, and plaque generations. In pAPPswe transfected SH-SY5Y cells, Ngb not only decreased the generation of Aß42, but also attenuated mitochondrial dysfunction and apoptosis through suppressing the activation of caspase-3, caspase-9 by Akt activating phosphorylation, which were restrained by phosphatidylinositol 3-kinase inhibitor (LY294002). Our data indicate the anti-apoptotic property of Ngb may play a neuroprotective role against AD.

Methylation status of WWOX gene promoter CpG islands in epithelial ovarian cancer and its clinical significance.

WW domain-containing oxidoreductase (WWOX) is a newly identified tumor suppressor gene that is associated with abnormal DNA methylation. The aim of this study was to evaluate the methylation status of CpG islands in the WWOX gene promoter region in cases of epithelial ovarian cancer and explore the correlation between the methylation status of the WWOX gene CpG islands and clinicopathological indices in patients with epithelial ovarian cancer. The methylation status of the WWOX gene CpG island was evaluated by methylation-specific polymerase chain reaction (MSP) in 48 patients with epithelial ovarian cancer, 18 patients with borderline epithelial ovarian tumors, 26 patients with epithelial benign tumors and 33 patients with normal ovarian tissues. Results showed that the rates of CpG island methylation in the WWOX gene promoter region in epithelial ovarian cancer tissues, borderline ovarian tumor tissues and benign ovarian tumor tissues were 43.75, 26.32 and 3.84%, respectively. The WWOX gene CpG islands were not methylated in normal ovarian tissues. The rate of CpG island methylation in epithelial ovarian cancer tissues was higher than that of other ovarian tissues and these differences were found to be statistically significant (P<0.01). The rate of CpG island methylation in the WWOX gene promoter region in late-stage (stage III and IV) epithelial ovarian cancer tissues was higher than that of early-stage (stage I and II) epithelial ovarian cancer tissues, and these differences were found to be statistically significant (P<0.05). In conclusion, epithelial ovarian cancer tissues showed CpG island hypermethylation in the WWOX gene promoter region, which may be an important mechanism leading to WWOX gene inactivation. Atypical methylation of WWOX gene is associated with the formation and progression of epithelial ovarian cancer, rendering it a potentially important indicator in the early diagnosis and prognosis of epithelial ovarian cancer.