CircNCOA4 knockdown attenuates OGD-induced neuron injury through miR-338-5p/PDE4B axis.

Circular RNAs (circRNAs) have been revealed to be involved in the pathology of acute ischemic stroke (AIS). Herein, we aimed to study the role and mechanism of circNCOA4 in ischemic stroke. The neuron-like cell line SK-N-SH of the experiment group was cultured in oxygen-glucose deprivation (OGD) condition. Cell viability and apoptosis were evaluated by cell counting kit-8 and flow cytometry. The oxidative damage and endoplasmic reticulum stress (ERS) were analyzed by measuring the production of reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD), and ERS-related markers. The binding between miR-338-5p and circNCOA4 or PDE4B (Phosphodiesterase 4B) was confirmed using dual-luciferase reporter and RIP assays. The commercial kit was used for exosome separation. The levels of circNCOA4 and PDE4B were increased, while miR-338-5p expression was decreased by OGD stimulation. OGD stimulation resulted in the apoptosis of neurons and induced oxidative damage and ERS, these effects were attenuated by circNCOA4 knockdown, while reinforced by circNCOA4 overexpression. Mechanistically, circNCOA4 acted as a sponge for miR-338-5p, and PDE4B was a target of miR-338-5p. MiR-338-5p inhibition reversed the neuroprotective effects of circNCOA4 silencing on neurons. Besides, miR-338-5p overexpression could abolish OGD-induced neuron injury, which was reversed by PDE4B upregulation. In addition, circNCOA4 was packaged into exosomes and showed potential diagnostic value for acute ischemic stroke (AIS) patients. CircNCOA4 has potential diagnostic value for AIS patients and promoted OGD-induced neuron injury via the miR-338-5p/PDE4B axis, providing a new insight into the pathology of AIS.

MDF-Net: A Multi-Scale Dynamic Fusion Network for Breast Tumor Segmentation of Ultrasound Images.

Breast tumor segmentation of ultrasound images provides valuable information of tumors for early detection and diagnosis. Accurate segmentation is challenging due to low image contrast between areas of interest; speckle noises, and large inter-subject variations in tumor shape and size. This paper proposes a novel Multi-scale Dynamic Fusion Network (MDF-Net) for breast ultrasound tumor segmentation. It employs a two-stage end-to-end architecture with a trunk sub-network for multiscale feature selection and a structurally optimized refinement sub-network for mitigating impairments such as noise and inter-subject variation via better feature exploration and fusion. The trunk network is extended from UNet++ with a simplified skip pathway structure to connect the features between adjacent scales. Moreover, deep supervision at all scales, instead of at the finest scale in UNet++, is proposed to extract more discriminative features and mitigate errors from speckle noise via a hybrid loss function. Unlike previous works, the first stage is linked to a loss function of the second stage so that both the preliminary segmentations and refinement subnetworks can be refined together at training. The refinement sub-network utilizes a structurally optimized MDF mechanism to integrate preliminary segmentation information (capturing general tumor shape and size) at coarse scales and explores inter-subject variation information at finer scales. Experimental results from two public datasets show that the proposed method achieves better Dice and other scores over state-of-the-art methods. Qualitative analysis also indicates that our proposed network is more robust to tumor size/shapes, speckle noise and heavy posterior shadows along tumor boundaries. An optional post-processing step is also proposed to facilitate users in mitigating segmentation artifacts. The efficiency of the proposed network is also illustrated on the "Electron Microscopy neural structures segmentation dataset". It outperforms a state-of-the-art algorithm based on UNet-2022 with simpler settings. This indicates the advantages of our MDF-Nets in other challenging image segmentation tasks with small to medium data sizes.

Case Report: Chemotherapy-free treatment with camrelizumab and anlotinib for elderly patients with KRAS and TP53 mutated advanced lung cancer.

Background: Lung cancer is a major public health issue and an enormous burden on society in China. Most lung cancers occur in elderly patients with non-small cell lung cancer (NSCLC), and many factors limit their treatment options. Chemotherapy-free therapy can avoid psychological fear, treatment pain, and adverse reactions caused by chemotherapy. Patients with non-small cell lung cancer with tumour protein p53 (TP53) gene mutations or Kirsten rat sarcoma viral oncogene homologue (KRAS) gene mutations tend to be more sensitive to anlotinib or programmed cell death protein 1 (PD-1) drugs. However, Kirsten rat sarcoma viral oncogene homologue is a proto-oncogene downstream of the epidermal growth factor receptor (EGFR) gene; therefore, if the Kirsten rat sarcoma viral oncogene homologue gene has an activating mutation, EGFR-targeted drug resistance may occur. Further studies are needed to explore whether patients with dual Kirsten rat sarcoma viral oncogene homologue and tumour protein p53 mutations can be treated with targeted immunotherapy without chemotherapy. Case presentation: A 74-year-old man was referred to the Lanzhou University Second Hospital due to chest tightness, shortness of breath, and weight loss for 2 months and was diagnosed with moderately to poorly differentiated adenocarcinoma. Laboratory examinations showed increased alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), cancer antigen (CA)-125, and CA199 levels, and gene sequencing indicated mutations in Kirsten rat sarcoma viral oncogene homologue and tumour protein p53. Immunohistochemical analysis showed positive PD-L1 and PD-1 expression. Peripheral blood immune checkpoint test using flow cytometry indicated that the PD-1 + CD8 levels were positive. After multi-disciplinary treatment, therapy with a combination of anlotinib and camrelizumab was initiated. Camrelizumab 200 mg was administered intravenously once every 3 weeks. Anlotinib 12 mg was administered orally daily before breakfast for 2 weeks with a week of rest in every cycle of 21 days. A reduction in alpha-fetoprotein, carcinoembryonic antigen, CA125, CA199, and CA724 levels was observed up to the first cycle, which decreased within the normal limits up to the second cycle and continued until the eighteenth cycle. The patient's chest tightness, shortness of breath, weight loss, and other symptoms significantly improved following treatment. Computed tomography imaging showed that the neoplastic lesion was dramatically reduced. The patient is currently being followed-up for more than 2 years to evaluate the duration of the response. Conclusion: Chemotherapy-free immunotherapy combined with targeted therapy is an effective treatment for advanced non-small cell lung cancer in elderly patients with Kirsten rat sarcoma viral oncogene homologue and tumour protein p53 mutations. Such therapies should be supported with further clinical studies with larger sample sizes.

Coagulation- and fibrinolysis-related genes for predicting survival and immunotherapy efficacy in colorectal cancer.

Background: Colorectal cancer (CRC) is a common cancer and has a poor prognosis. The coagulation system and fibrinolysis system are closely related to the progression of malignant tumors and is also related to the immunotherapy of malignant tumors. Herein, we tried to predict survival and the immunotherapy effect for patients with CRC using a novel potential prognostic model. Methods: Through online data of TCGA and GEO, we screened significantly differentially expressed genes (DEGs) to construct a prognostic model, followed by obtaining immune-related genes (IRGs) from the ImmPort database and coagulation- and fibrinolysis-related genes (CFRGs) from the GeneCards database. The predictive power of the model is assessed by Kaplan-Meier survival curves as well as the time-dependent ROC curve. Moreover, univariate and multivariate analyses were conducted for OS using Cox regression models, and the nomogram prognostic model was built. In the end, we further studied the possibility that CXCL8 was associated with immunocyte infiltration or immunotherapy effect and identified it by immunohistochemistry and Western blot. Results: Five DEGs (CXCL8, MMP12, GDF15, SPP1, and NR3C2) were identified as being prognostic for CRC and were selected to establish the prognostic model. Expression of these genes was confirmed in CRC samples using RT-qPCR. Notably, those selected genes, both CFRGs and IRGs, can accurately predict the OS of CRC patients. Furthermore, CXCL8 is highly correlated with the tumor microenvironment and immunotherapy response in CRC. Conclusion: Overall, our established IRGPI can very accurately predict the OS of CRC patients. CXCL8 reflects the immune microenvironment and reveals the correlation with immune checkpoints among CRC patients.

Interaction between gut microbiota and tumour chemotherapy

Chemotherapeutic drugs play an important role in the treatment of cancer, but the individual differences of patients' sensitivity to chemotherapeutic drugs and the drug resistance of chemotherapeutic drugs have always been a thorny problem in clinical treatment. In recent years, with the progress in research on human microbiota, gut microbiome plays an increasingly important role in the diagnosis and treatment of diseases. Studies have shown that gut microbiota can regulate the tumour microenvironment and affect the efficacy and toxicity of chemotherapy through a variety of mechanisms. This paper focuses on the specific mechanism that gut microbiota uses to influence chemotherapy and the potential therapeutic effect of supplementing with probiotics, to provide an important basis for individualised treatment strategies to be used when treating malignant tumours (AU)

Interaction between gut microbiota and tumour chemotherapy.

Chemotherapeutic drugs play an important role in the treatment of cancer, but the individual differences of patients' sensitivity to chemotherapeutic drugs and the drug resistance of chemotherapeutic drugs have always been a thorny problem in clinical treatment. In recent years, with the progress in research on human microbiota, gut microbiome plays an increasingly important role in the diagnosis and treatment of diseases. Studies have shown that gut microbiota can regulate the tumour microenvironment and affect the efficacy and toxicity of chemotherapy through a variety of mechanisms. This paper focuses on the specific mechanism that gut microbiota uses to influence chemotherapy and the potential therapeutic effect of supplementing with probiotics, to provide an important basis for individualised treatment strategies to be used when treating malignant tumours.

The Shoulder Charcot Arthropathy and Secondary to Hemangioblastoma in Conus Medullari with Syringomyelia.

Charcot arthropathy of the shoulder joint is usually referred to as shoulder joint involvement of Charcot arthropathy, which is a chronic, degenerative, destructive condition resulting from decreasing or loss of sensorial innervation. To date, several reports have described the shoulder Charcot arthropathy, caused by malformations of the occipital region with syringomyelia; but nobody has reported the shoulder Charcot arthropathy secondary to hemangioblastoma in conus medullari with syringomyelia. Therefore, we report a 32-year male patient who was diagnosed with Charcot arthropathy of the shoulder joint, which was misdiagnosed as a soft tissue tumor and treated surgically. After the operation, the whole-spine MRI examination was performed as a presenting feature of hemangioblastoma in conus medullari with syringomyelia. Key Words: Arthropathy, Neurogenic, Hemangioblastoma, Syringomyelia.

BUB1 predicts poor prognosis and immune status in liver hepatocellular carcinoma.

Accurate assessment of the tumour immune microenvironment promotes individualized immunotherapy regimens and screens dominant populations suitable for immunotherapy. Therefore, potential molecular markers were investigated to make an overall assessment of the immune microenvironment status of liver hepatocellular carcinoma (LIHC). In this study, a total of 121 differentially expressed genes (DEGs) were identified, and DEGs were enriched in the epithelial-mesenchymal transition, hypoxia, myogenesis, and p53 pathways. A total of 20 hub genes were selected and a strong correlation was identified between these hub genes and prognosis. The expression of budding uninhibited by benzimidazoles 1 (BUB1) was found to be upregulated in LIHC and was strongly related to immune cells and immune checkpoint molecule expression. Immunohistochemistry (IHC) indicated that BUB1 expression was higher in LIHC tissues than in normal liver tissues. BUB1 knockdown resulted in reduced proliferation and vertical migration ability of LIHC cells, and reduced the expression of phospho-SMAD family member 2 and phospho-SMAD family member 3 proteins. IHC showed that BUB1 expression was accompanied by immune cell infiltration into LIHC tissues. These results suggest that BUB1 may serve as a potential prognostic biomarker for LIHC and as an indicator of its immune status.

A Cross-Sectional Study on the Cross-Talk of the COVID-19-Related Degree of Loneliness and the Etiological Factors Among the Elderly in Central China.

BACKGROUND: The outbreak of COVID-19 has undoubtedly influenced the normal lifestyle of people worldwide, including the Chinese population. This study attempted to do supplementary research to the current situation of loneliness as well as the related risk factors among the elderly in the province in central Chinese during the COVID-19. METHODS: We conducted a cross-sectional study in one of the central Chinese provinces (Henan Province) from December 2020 to March 2021 using a multistage sampling method, and 568 elderly people without cognitive impairment were interviewed. The UCLA Loneliness Scale, Pittsburgh Sleep Quality Index (PSQI), Physical Activity Rating Scale (PARS-3), and Quality of Life Questionnaire SF-36 were adopted to collect information. We used univariate and multivariate logistic regressions to analyze the factors resulting in severe loneliness among the elderly with seldom or regular participation in physical exercises. RESULTS: During the epidemic in central China, the elderly suffering from loneliness syndrome accounted for 34.2%, of which 15.5% were severely lonely. Risk factors for severe loneliness were quality of life (OR: 7.129), sleep quality (OR: 3.778), seldom exercise (OR: 4.170), poor economic status (OR: 1.769), and negative attitude toward the prospects for the epidemic control (OR: 4.033). By grouping the participants in terms of physical activity, we found that the quality of life (OR:5.778) was a significant risk factor than sleep quality (OR:2.939) in the seldom exercise group, while the only risk factor in the regular exercise group was the quality of life (OR: 5.021). CONCLUSION: There was an increase in the degree of loneliness among the elderly during the epidemic, and physical activity played an active role in relieving the severe loneliness of the elderly. Therefore, for the sake of the elderly, regular participation in physical exercises should be encouraged during the duration of the epidemic.

Efficient Estimation of Permeate Flux of Asymmetric Ceramic Membranes for Vacuum Membrane Distillation.

Ceramic membranes have the advantages of high mechanical strength and thermal stability and are promising candidates for membrane distillation. Ceramic membranes are generally designed to have a multilayer structure with different pore sizes to create a high liquid entry pressure and obtain a high permeability. However, these structural characteristics pose significant difficulties in predicting permeate flux in a ceramic membrane contactor for vacuum membrane distillation (VMD). Here, a modeling approach was developed to simulate the VMD process and verified by comparing the simulated results with the experimental data. Furthermore, correlations are proposed to simplify the calculations of permeate flux for VMD using asymmetric ceramic membranes by assuming those multilayers to be an effectively quasi-symmetric layer and by introducing a correction factor. The simulation results indicated that this simplified correlation was effective and enabled a quick estimation of the effect of membrane parameters on permeate flux.

Current research developments of patient-derived tumour xenograft models (Review).

Patient-derived tumor xenograft (PDTX) models are established by transferring patient tumors into immunodeficient mice. In these murine models, the characteristics of the primary tumor are retained, including the microenvironment of tumor cell growth and histopathology. Due to this, it has become the most reliable in vivo human cancer model. However, the success rates differ by type of tumor, site of transplantation and tumor aggressiveness. Subcutaneous transplantation is a standard method for PDTX, and subrenal capsule transplantation improves the engraftment rate. Recently, PDTX models are frequently used in the fields of precision medicine, predictive biomarkers, evaluation of drug efficacy and preclinical research on tumor immunotherapeutic drugs. The aim of the present article was to review the establishment, clinical applications and limitations of the PDTX model in tumor research.

Identification of Seven-Gene Hypoxia Signature for Predicting Overall Survival of Hepatocellular Carcinoma.

BACKGROUND: Hepatocellular carcinoma (HCC) is ranked fifth among the most common cancer worldwide. Hypoxia can induce tumor growth, but the relationship with HCC prognosis remains unclear. Our study aims to construct a hypoxia-related multigene model to predict the prognosis of HCC. METHODS: RNA-seq expression data and related clinical information were download from TCGA database and ICGC database, respectively. Univariate/multivariate Cox regression analysis was used to construct prognostic models. KM curve analysis, and ROC curve were used to evaluate the prognostic models, which were further verified in the clinical traits and ICGC database. GSEA analyzed pathway enrichment in high-risk groups. Nomogram was constructed to predict the personalized treatment of patients. Finally, real-time fluorescence quantitative PCR (RT-qPCR) was used to detect the expressions of KDELR3 and SCARB1 in normal hepatocytes and 4 HCC cells. The expressions of SCARB1 in hepatocellular carcinoma tissue in 46 patients were detected by immunohistochemistry, and the correlation between its expressions and disease free survival of patient was calculated. RESULTS: Through a series of analyses, seven prognostic markers related to HCC survival were constructed. HCC patients were divided into the high and low risk group, and the results of KM curve showed that there was a significant difference between the two groups. Stratified analysis, found that there were significant differences in risk values of different ages, genders, stages and grades, which could be used as independent predictors. In addition, we assessed the risk value in the clinical traits analysis and found that it could accelerate the progression of cancer, while the results of GSEA enrichment analysis showed that the high-risk group patients were mainly distributed in the cell cycle and other pathways. Then, Nomogram was constructed to predict the overall survival of patients. Finally, RT-qPCR showed that KDELR3 and SCARB1 were highly expressed in HepG2 and L02, respectively. Results of IHC staining showed that SCARB1 was highly expressed in cancer tissues compared to adjacent normal liver tissues and its expression was related to hepatocellular carcinoma differentiation status. The Kaplan-Meier survival showed a poor percent survival in the SCARB1 high group compared to that in the SCARB1 low group. CONCLUSION: This study provides a potential diagnostic indicator for HCC patients, and help clinicians to deepen the comprehension in HCC pathogenesis so as to make personalized medical decisions.

Permeability and Stability of Hydrophobic Tubular Ceramic Membrane Contactor for CO2 Desorption from MEA Solution.

Ceramic membrane contactors hold great promise for CO2 desorption due to their high mass transfer area as well as the favorable characteristics of ceramic materials to resist harsh operating conditions. In this work, a hydrophobic tubular asymmetric alpha-alumina (α-Al2O3) membrane was prepared by grafting a hexadecyltrimethoxysilane ethanol solution. The hydrophobicity and permeability of the membrane were evaluated in terms of water contact angle and nitrogen (N2) flux. The hydrophobic membrane had a water contact angle of ~132° and N2 flux of 0.967 × 10-5 mol/(m2∙s∙Pa). CO2 desorption from the aqueous monoethanolamine (MEA) solution was conducted through the hydrophobic tubular ceramic membrane contactor. The effects of operating conditions, such as CO2 loading, liquid flow rate, liquid temperature and permeate side pressure, on CO2 desorption flux were investigated. Moreover, the stability of the membrane was evaluated after the immersion of the ceramic membrane in an MEA solution at 373 K for 30 days. It was found that the hydrophobic α-Al2O3 membrane had good stability for CO2 desorption from the MEA solution, resulting in a <10% reduction of N2 flux compared to the membrane without MEA immersion.

Quantification of the CM-Dil-labeled human umbilical cord mesenchymal stem cells migrated to the dual injured uterus in SD rat.

BACKGROUND: Human umbilical cord mesenchymal stem cell (hUC-MSC) therapy is considered as a promising approach in the treatment of intrauterine adhesions (IUAs). Considerable researches have already detected hUC-MSCs by diverse methods. This paper aims at exploring the quantitative distribution of CM-Dil-labeled hUC-MSCs in different regions of the uterus tissue of the dual injury-induced IUAs in rats and the underlying mechanism of restoration of fertility after implantation of hUC-MSCs in the IUA model. METHODS: In this study, we investigated the quantification of the CM-Dil-labeled hUC-MSCs migrated to the dual injured uterus in Sprague Dawley rats. Additionally, we investigated the differentiation of CM-Dil-labeled hUC-MSCs. The differentiation potential of epithelial cells, vascular endothelial cells, and estrogen receptor (ER) cells were assessed by an immunofluorescence method using CK7, CD31, and ERα. The therapeutic impact of hUC-MSCs in the IUA model was assessed by hematoxylin and eosin, Masson, immunohistochemistry staining, and reproductive function test. Finally, the expression of TGF-ß1/Smad3 pathway in uterine tissues was determined by qRT-PCR and Western blotting. RESULTS: The CM-Dil-labeled cells in the stroma region were significantly higher than those in the superficial myometrium (SM) (71.67 ± 7.98 vs. 60.92 ± 3.96, p = 0.005), in the seroma (71.67 ± 7.98 vs. 23.67 ± 8.08, p = 0.000) and in the epithelium (71.67 ± 7.98 vs. 4.17 ± 1.19, p = 0.000). From the 2nd week of treatment, hUC-MSCs began to differentiate into epithelial cells, vascular endothelial cells, and ER cells. The therapeutic group treated with hUC-MSCs exhibited a significant decrease in fibrosis (TGF-ß1/Smad3) as well as a significant increase in vascularization (CD31) compared with the untreated rats. CONCLUSION: Our findings suggested that the distribution of the migrated hUC-MSCs in different regions of the uterine tissue was unequal. Most cells were in the stroma and less were in the epithelium of endometrium and gland. Injected hUC-MSCs had a capacity to differentiate into epithelial cells, vascular endothelial cells, and ER cells; increase blood supply; inhibit fibration; and then restore the fertility of the IUA model.

Gli1 regulates stemness characteristics in gastric adenocarcinoma.

BACKGROUND: Glioma-associated oncogene homolog 1 (Gli1), affects the progression and the stemness characteristics of malignant carcinoma. The aim of the present study was to identify the relation between Glioma-associated oncogene homolog 1 (Gli1) and stemness and determine its clinical significance in gastric adenocarcinoma (GA). We investigated Gli1 expression and its correlation with other stemness-associated proteins in 169 GA samples and 5 GA cell lines. METHODS: To elucidate the role of Gli1 in the clinicopathological significance and stemness of GA, tissues samples from 169 GA patients were collected for immunohistochemistry (IHC). Additionally, MKN74, MKN28, NCI-N87, SNU638, AGS cells were collected for western blotting, MKN28 cells were collected for spheroid formation assay. RESULTS: Results showed that Gli1 expression was closely related to tumor grade, primary tumor (pT) stage, distant metastasis, clinical stage, gross type, microvessel density, and shorter overall survival (OS). Cox regression analysis verified that Gli1 was an independent prognostic factor for OS. Furthermore, Gli1 expression correlated with the expression of stemness-related genes, CD44, LSD1, and Sox9. Gli1 inhibitor GANT61 significantly decreased the expression of CD44 and LSD1, and spheroid formation ability of the MKN28 cells. CONCLUSIONS: In conclusion, Gli1 may be a poor prognostic indicator and a potential cancer stemness-related protein in GA.

Exosomes Derived from Adipose Mesenchymal Stem Cells Restore Functional Endometrium in a Rat Model of Intrauterine Adhesions.

Intrauterine adhesion (IUA) caused by endometrial injury is one of the important causes of infertility in women of reproductive age and requires advanced treatment strategies. Increasing evidence suggests that the therapeutic effects of mesenchymal stem cells (MSC) mainly depend on their capacity to secrete paracrine factors and are mediated by MSC-derived exosomes. This study aimed to identify exosomes derived from adipose-derived mesenchymal stem cells (ADSC-exo) and explore the therapeutic potential in IUA rat models. ADSC-exo exhibited classic cup-shaped morphology with a positive expression of Alix and CD63 and were mainly concentrated at 109.5 nm. In IUA model, treatment with ADSC-exo maintained normal uterine structure, promoted endometrial regeneration and collagen remodeling, and enhanced the expression of integrin-ß3, LIF, and VEGF. An improved receptivity of the regenerated endometrium was confirmed. Our findings demonstrated that ADSC-exo promoted endometrial regeneration and fertility restoration. It suggested that topical administration of ADSC-exo in uterus could be a promising strategy for patients suffering severe intrauterine adhesions and infertility.

LETM1 is a potential cancer stem-like cell marker and predicts poor prognosis in colorectal adenocarcinoma.

The leucine zipper-EF-hand containing transmembrane protein 1 (LETM1) is highly expressed in many human malignancies and is correlated with poor prognosis. However, LETM1 has rarely been explored as a cancer stem-like cell marker for the prognostic evaluation of colorectal adenocarcinoma (CRA). Herein, we assessed the expression of LETM1 and its relationship with cancer stemness genes, cell cycle markers, PI3K/Akt/NFκB signaling pathway genes, and HIF1α in 102 paraffin-embedded CRA tissue samples using immunohistochemistry (IHC). Additionally, we further confirmed the correlation between LETM1 and cancer stemness genes in CRA cell lines using immunofluorescence (IF) imaging and Western blotting. LETM1 expression was remarkably upregulated in human fetal sagittal sections and CRA tissues. The expression of LETM1 in CRA tissue was correlated with clinical stage, lymph node metastasis, distant metastasis, and microvessel density. LETM1 expression was significantly associated with lower overall survival and disease-free survival. Moreover, the expression of LETM1 positively correlated with SOX9, LSD1, CD44, CD133, LGR5, SOX2, and HIF1α. IF revealed that LETM1 co-localized with CD44, SOX9, and LGR5 in HCT116. Moreover, LETM1 expression was also strongly linked to the expression of cell cycle regulators (cyclinD1, CDK4, p27) and PI3K/Akt/NFκB pathway genes (pPI3K-p85, pAkt-Ser473, pAkt-Thr308, pNFκB-p65). LETM1 may therefore be a cancer stem-like cell marker and an indicator of poor prognosis in patients with CRA.

Tenascin-C is involved in promotion of cancer stemness via the Akt/HIF1ɑ axis in esophageal squamous cell carcinoma.

Although tenascin-C (TNC), an extracellular matrix protein, has been shown to be widely expressed in stromal fibroblasts in various cancers, the role of its expression in esophageal squamous cell carcinoma (ESCC) cells remains unclear. Using immunohistochemistry, we investigated the expression of cancer stem-like cell (CSC) markers, epithelial-to-mesenchymal transition (EMT)-related genes, and the Akt/hypoxia-inducible factor-1α (HIF1α) signal pathway in ESCC tissue specimens from 154 patients. We further addressed the effects of TNC on the Akt/HIF1α axis and its putative association with cancer stemness in several ESCC cell lines by immunofluorescence imaging and western blot analysis. Our data suggest that TNC expression was positively correlated with the expression of the CSC marker SOX2 (p = .002), and TNC-expressing cancer cells expressed SOX2 in ESCC tissues. Moreover, TNC expression was strongly associated with EMT-related gene Snail (p = .022) and positively correlated with pAkt-Ser473 (p = .004) and HIF1α (p = .003). Furthermore, TNC-silencing down-regulated the expression of CSC marker SOX2 (p < .001) and EMT-related marker Snail (p < .001). The Akt inhibitor Perifosine inhibited the protein expression of pAkt-Ser473, Akt, HIF1α, and TNC in TE10 (an ESCC cell line) cells. Short-term exposure of TE10 cells to cobalt chloride caused an increase in protein expression of HIF1α, TNC, and SOX2 in a time-dependent manner. Taken together, these results suggest that TNC may enhance the cancer stem-like properties and promote EMT-like changes via the Akt/HIF1α axis.

The role of Tenascin-C and Twist1 in gastric cancer: cancer progression and prognosis.

The aim of the present study was to identify the relation between Tenascin-C (TNC) and Twist1 and determine their clinical significance in gastric cancer (GC). We analyzed the expression of TNC and Twist1 in 159 GC samples and in 91 non-tumor samples using immunohistochemistry. In this study, TNC expression in stromal fibroblasts of GC was remarkably higher than non-tumor gastric lesions. The expression of TNC in GC stromal fibroblasts was significantly associated with pT stage, lymph node metastasis, distant metastasis. Twist1 expression in stromal fibroblasts of GC was remarkably higher than non-tumor gastric lesions. Twist1 expression in the stromal fibroblasts of GC was associated with tumor size, lymph node metastasis, and clinical stage. Furthermore, TNC expression levels in GC stromal fibroblasts were positively associated with Twist1. The simultaneous expression of TNC and Twist1 was significantly higher in stromal fibroblasts of GC than in noncancerous tissues. The simultaneous expression of TNC and Twist1 in GC stromal fibroblasts was positively associated with tumor location, pT stage, lymph node metastasis and clinical stage. Moreover, patients with co-expression of TNC and Twist1 had a poorer prognosis than either TNC or Twist1 positive in GC. Our study revealed that the simultaneous expression of TNC and Twist1 indicated the poorer prognosis of GC. Co-targeting TNC and Twist1 confer significant clinical advantage, which offers a novel therapeutic target in GC.

GLI1 promotes cancer stemness through intracellular signaling pathway PI3K/Akt/NFκB in colorectal adenocarcinoma.

The role of Hedgehog (HH)/ glioma-associated oncogene homolog 1 (GLI1) pathway has been implicated in a variety of cancer entities, and the targeted pathway inhibition mediated by GLI1 is of therapeutic relevance. However, its oncogenicity and cross-talks with other cancer pathways including PI3K/Akt/NFκB, which modulates the HH/GLI1 signal strength, have rarely been explored in colorectal adenocarcinoma. We assessed the expression of GLI1 and its relationship with other cancer stemness genes, cell cycle markers, epithelial-mesenchymal transition (EMT), PI3K/Akt/NFκB signaling pathway genes, and HIF1α in 100 paraffin-embedded colorectal adenocarcinoma tissue samples using immunohistochemistry. We further addressed the effect of GLI1 on EMT, cell cycle, and its putative interaction with the PI3K/Akt/NFκB cascade in colorectal adenocarcinoma cell lines. The expression of GLI1 in colorectal adenocarcinoma tissues was found to correlate with the clinical stages, and distant metastasis. Moreover, GLI1 was found to be an independent predictor of poor overall survival and disease-free survival in colorectal adenocarcinoma. GLI1-expressing cancer cells also expressed their representative cancer stem-like cell (CSC) markers (SOX9 and CD133), as well as HIF1α. GLI1 expression was also strongly linked to EMT-related and PI3K/Akt/NFκB signaling genes. Downregulation of GLI1 by inhibitor treatment in colorectal adenocarcinoma cell lines resulted in reduced expression of CSC markers, cell clonogenicity, S-phase subpopulations, as well as the migration and invasion ability. Importantly, Akt inhibitor Perifosine significantly inhibited the expression of pAkt and GLI1 in colorectal adenocarcinoma cells. Combination of GLI1 inhibitor GANT61 and NFκB p65 inhibitor QZN exhibited much higher inhibition compared to using any of them individually on colorectal adenocarcinoma cells. We suggested that GLI1 may be a novel stem cell marker, and cancer stemness was activated via PI3K/Akt/NFκB pathway. In addition, co-targeting GLI1 and PI3K/Akt/NFκB signaling simultaneously might provide an alternative therapeutic strategy for colorectal adenocarcinoma patients.