ISG15 is a potential therapeutic target for osteosarcoma: a comprehensive analysis based on bioinformatics and in vitro experiments.

BACKGROUND: The expression of aberrant interferon-stimulated gene 15 (ISG15) is connected with various human diseases, including cancer. ISG15 is involved in tumor formation and metastasis. However, its role in osteosarcoma is uncertain. METHODS: ISG15 expression in pan-cancer from RNA Sequencing data were obtained from The Cancer Genome Atlas (TCGA) and Genotype Tissue Expression (GTEx) databases. The relationship between ISG15 expression and prognosis was assessed through TCGA clinical survival data. Immunohistochemistry (IHC) images of ISG15 were retrieved using the Human Protein Atlas to analyze the differences in selected normal and tumor tissues. Gene enrichment analysis and signaling pathway analysis were used to assess the potential role of ISG15 in sarcoma, and the correlation between ISG15 expressions and immune cell infiltration levels was estimated by immune infiltration analysis. The expression levels of ISG15 were assessed by qRT-PCR and IHC. Colony formation, wound healing assay and transwell assay were used to detect the effects of ISG15 on the biological behaviors of osteosarcoma cells. The correlation between ISG15 levels and CD8+/CD68+ cells was further examined by double-labeled immunofluorescence. The chemotactic effect of ISG15 on CD8+/CD68+ cells was demonstrated by chemotactic experiments and flow cytometry. RESULTS: ISG15 was highly expressed in most cancers, while high ISG15 expression was significantly correlated with poor overall survival. Gene enrichment analysis in sarcoma suggested that antigen processing and presentation might be involved in the oncogenic mechanism of ISG15. Further immune infiltration analysis showed that high ISG15 expression might reflect the infiltration level of certain immune cells. Additionally, our verification showed that ISG15 was significantly related to the occurrence and metastasis of osteosarcoma, and knockdown of ISG15 significantly altered cell biological behavior, resulting in decreased proliferation, migration and invasion capabilities of osteosarcoma cells. The high expression of ISG15 in osteosarcoma tissue was associated with a high level of CD68+ immune cell infiltration while a low level of CD8+ T cell infiltration. CD68+ immune cells were recruited in vitro by overexpression of ISG15, which on the contrary could weaken the chemotaxis of CD8+ T cells. CONCLUSION: High ISG15 expression is an inherent feature of osteosarcoma and triggers tumorigenesis and metastasis by regulating tumor immunogenicity. ISG15 is expected to be the target of osteosarcoma treatment.

LncRNA TEX41 regulates autophagy by increasing Runx2 expression in lung adenocarcinoma bone metastasis.

OBJECTIVE: To investigate the mechanism underlying the role of TEX41 in lung adenocarcinoma (LUAD) bone metastasis (BM). METHODS: We analyzed the biological functions and molecular mechanisms of TEX41 using bioinformatics. TEX41 and Runx2 expressions were measured in clinical tissue samples and cell lines by quantitative PCR. The effects of TEX41 on LUAD cell proliferation, migration, invasion and metastasis as well as its mechanism of action were investigated. Fluorescence in-situ hybridization (FISH) was performed to determine TEX41 and Runx2 colocalization. Subcutaneous tumor growth and BM were evaluated in nude mice by X-ray and hematoxylin and eosin (HE) staining. RESULTS: TEX41 was dramatically increased in LUAD BM tissue, indicating a poorer prognosis in patients with LUAD and BM. TEX41 knockdown suppressed the migration and metastasis of LUAD cells, whereas TEX41 overexpression promoted these processes. Data from X-ray and HE staining showed that TEX41 supported the BM in LUAD. TEX41 overexpression induced autophagy in LUAD cells, as demonstrated by changes in autophagy markers. Results of FISH showed that TEX41 and Runx2 colocalized in the nucleus, and Runx2 expression was regulated by TEX41. The effects of TEX41 on LUAD cell migration, invasion, metastasis and autophagy were counteracted by Runx2 inhibition. Moreover, the role of TEX41 in the metastasis was partially dependent on autophagy, and phosphoinositide 3-kinase (PI3K)-AKT might be the major signaling pathway involved in TEX41-regulated autophagy. CONCLUSION: TEX41 promotes autophagy in LUAD cells by upregulating Runx2 to mediate LUAD migration, invasion and BM.

The formation of higher alcohols in rice wine fermentation using different rice cultivars.

Higher alcohols are closely related to the flavor and safety of rice wine. The formation of n-propanol, isobutanol, isoamyl alcohol, and phenylethanol during rice wine fermentations was for the first time investigated in this study among 10 rice cultivars from two main production regions. Rice wine made from Yashui rice, the long-grain non-glutinous rice from Guizhou, produced the highest yields of higher alcohols (487.45 mg/L), and rice wine made from five glutinous rice cultivars produced the lowest yields of higher alcohols (327.45-344.16 mg/L). An extremely strong correlation was found between the starch in rice and higher alcohols in rice wine. Further analysis first showed that the former fermentation period was key for the nutrient consumption and higher alcohol formation, with more than 55% of glucose being consumed and more than 75% of higher alcohols being synthesized in 48 h. Correlation analysis confirmed the strong correlation between nutrient consumption and higher alcohol formation including valine-isobutanol (coefficient higher than 0.8 in seven rice cultivars and higher than 0.6 in three rice cultivars), glucose-isoamyl alcohol (coefficient higher than 0.8 in five rice cultivars and higher than 0.6 in the other five rice cultivars), and glucose-phenylethanol (coefficient higher than 0.8). The correlation of threonine-n-propanol, leucine-isoamyl alcohol, phenylalanine-phenylethanol, glucose-n-propanol, and glucose-isobutanol varied among the rice wines made from 10 rice cultivars. RT-qPCR analysis on five target genes verified the variation caused by different rice cultivars. this study for the first time reported the special formation pattern of higher alcohols during rice wine fermentation, emphasizing the early contribution of glucose metabolism on the formation of isobutanol. This study highlighted the significance of rice selection for making rice wine with good quality and provided theoretical references for the control of higher alcohols, especially in the former period of rice wine fermentation.

MicroRNA-106a regulates autophagy-related cell death and EMT by targeting TP53INP1 in lung cancer with bone metastasis.

Bone metastasis is one of the most serious complications in lung cancer patients. MicroRNAs (miRNAs) play important roles in tumour development, progression and metastasis. A previous study showed that miR-106a is highly expressed in the tissues of lung adenocarcinoma with bone metastasis, but its mechanism remains unclear. In this study, we showed that miR-106a expression is dramatically increased in lung cancer patients with bone metastasis (BM) by immunohistochemical analysis. MiR-106a promoted A549 and SPC-A1 cell proliferation, migration and invasion in vitro. The results of bioluminescence imaging (BLI), micro-CT and X-ray demonstrated that miR-106a promoted bone metastasis of lung adenocarcinoma in vivo. Mechanistic investigations revealed that miR-106a upregulation promoted metastasis by targeting tumour protein 53-induced nuclear protein 1 (TP53INP1)-mediated metastatic progression, including cell migration, autophagy-dependent death and epithelial-mesenchymal transition (EMT). Notably, autophagy partially attenuated the effects of miR-106a on promoting bone metastasis in lung adenocarcinoma. These findings demonstrated that restoring the expression of TP53INP1 by silencing miR-106a may be a novel therapeutic strategy for bone metastatic in lung adenocarcinoma.

Profiling of Fungal Diversity and Fermentative Yeasts in Traditional Chinese Xiaoqu.

To increase the safety and quality of baijiu and rice wine in China, controlling the use of traditional Xiaoqu by studying the beneficial yeasts present has recently been considered. The fungal diversity of six Chinese Xiaoqu including five traditional and one commercial samples was investigated to screen fermentative yeasts with low yields of higher alcohols. A high throughput sequencing approach detected fifteen fungal species with relative abundance higher than 1%, and displayed dissimilarities of fungal diversity among Xiaoqu samples. The 15 fungal species were composed of 11 filamentous fungi with Rhizopus arrhizus as the most common specie and four yeast species, containing Hyphopichia burtonii, Saccharomyces cerevisiae, Saccharomycopsis fibuligera, and Saccharomycopsis malanga. Classic culture-dependent approaches, including 5.8S-ITS-RFLP analysis and D1/D2 sequencing of the 26S rRNA gene, identified nine yeast species in the five traditional Chinese Xiaoqu. In addition to the four yeast species also detected by high throughput sequencing approach, the other five yeast species isolated were Clavispora lusitaniae, Cryptococcus neoformans, Komagataella pastoris, Trichosporon asahii, and Wickerhamomyces anomalus. Further micro-fermentations of rice wine were performed using 19 single yeast isolates, and after the fermentation of rice wine, higher alcohols and ethanol were analyzed by gas chromatography. Two yeast strains, Saccharomyces cerevisiae FBKL2.8022 and Wickerhamomyces anomalus FBKL2.8023, were found to have low yields of higher alcohols and could produce 11.70%vol and 7.10%vol ethanol separately. This study for the first time, to the best of our knowledge, explored the fungal resources in traditional Xiaoqu from different regions of Guizhou, China. The screened S. cerevisiae and W. anomalus strains could be used to establish specific starters to promote the standardization of the production of baijiu and rice wine.

Effects of different levels of TGF-ß expression and tumor cell necrosis rates in osteosarcoma on the chemotherapy resistance of osteosarcoma.

PURPOSE: The clinical significance of transforming growth factor ß (TGF-ß) and tumor cell necrosis rate (TCNR) in the expression of osteosarcoma and its effects of chemotherapy resistance on osteosarcoma were explored. PATIENTS AND METHODS: 94 cases of neoadjuvant chemotherapy osteosarcoma patients at the Third Affiliated Hospital of Kunming Medical University between January 2014 and January 2019 were collected. Samples tested for TGF-ß were collected before chemotherapy, the tumor cell necrosis rate of pathological samples before and after chemotherapy was determined. Others analyzed covariates included 12 prognostic factors that may be associated with chemotherapy resistance in previous studies: age, BMI, initial diagnosis time (The time from symptom onset to first medical attention), KPS score, initial tumor size, lymphocytes/leukocytes rate (LWR), neutrophils/lymphocytes rate (NLR), albumin, aspartate transaminase (AST), low density lipoprotein (LDL), blood urea nitrogen (BUN), alkaline phosphatase (ALP), the endpoints included progression-free survival (PFS) and overall survival (OS), response evaluation criteria in solid tumours by RECIST guideline (version 1.1). RESULT: 1. A total of 94 cases were examined for expression of TGF-ß in pathological specimens, 45 cases were TGF-ß high expression (47.9%) and 49 cases were TGF-ß low expression (52.1%); 2. The BMI, LDL, ALP, NLR in TGF-ß high expression group was significantly increased compared to TGF-ß low expression group; the Initial diagnosis time, KPS in TGF-ß high expression group was significantly decreased compared to TGF-ß low expression group, all P < 0.05; 3. Effect of chemotherapy was positively with positive cell rate (P < 0.01 r = 0.337) and TGF-ß total score (P < 0.0001 r = 0.635), while effect of chemotherapy was no correlation with degree of dyeing score (P > 0.05); there was significant difference in change from baseline after chemotherapy between TGF-ß high expression group and TGF-ß low expression group (P = 0.045); 4. Median OS 61.4 months in the TGF-ß high expression group, median OS 68.1 months in the TGF-ß low expression group, one-year survival rate, there was statistically significant difference in two groups (P = 0.045); median PFS 44.8 months in the TGF-ß high expression group, median PFS 56.2 months in the TGF-ß low expression group, There was no statistically significant difference in two groups (P > 0.05); 5. A total of 92 cases were examined for TCNR after chemotherapy, 62 were TCNR ≤ 90% (67.4%), 30 were TCNR > 90% (32.6%); 6. the Initial diagnosis time, KPS, in TCNR > 90% group was significantly increased compared to TCNR ≤ 90% group; the initial tumor size, BUN, ALP in TCNR > 90% group was significantly decreased compared to TCNR ≤ 90% group, all P < 0.05; 7. TCNR was negatively correlated with the change from baseline after chemotherapy (P < 0.001 r = -0.411); there was no statistically significant difference between TCNR > 90% group and TCNR ≤ 90% group in change from baseline after chemotherapy (P > 0.05); 8. Median OS 67.8 months in the TCNR > 90% group, median OS 61.7 months in the TCNR ≤ 90% group, there was statistically significant difference between two groups (P = 0.040); median PFS 57.4 months in the TCNR > 90% group, median PFS 40.5 months in the TCNR ≤ 90% group, there was statistically significant difference between two groups (P = 0.036); 9. TGF-ß total score was negatively correlated with TCNR (P < 0.001 r = -0.571). CONCLUSION: The results of this study suggested that the higher expression of TGF-ß, the lower expression of TCNR, which more likely to induce chemotherapy resistance among patients with osteosarcoma and lead to poor prognosis.

The characteristics of lung cancer in Xuanwei County: A review of differentially expressed genes and noncoding RNAs on cell proliferation and migration.

The occurrence of lung cancers is the highest in Xuanwei County, Yunnan province, China, especially among nonsmoking women. Domestic combustion of smoky coal induces serious indoor air pollution and is considered to be the main cause of human lung cancers. The occurrence of lung cancer in Xuanwei County has unique characteristics, such as the high morbidity in nonsmoking women or people with no family history. In the present review, we summarize advances in identification of differentially expressed genes, regulatory lncRNAs and miRNAs in cell proliferation and migration of lung cancers in Xuanwei County. Moreover, several regulatory differentially expressed genes (DEGs) or noncoding RNAs have diagnostic and prognostic significance for lung cancers in Xuanwei County and have the potential to serve as biomarkers.