Identification and immunological characteristics of anoikis-associated molecular clusters in lung adenocarcinoma.

Anoikis plays a crucial role in the progression, prognosis, and immune response of lung adenocarcinoma (LUAD). However, its specific impact on LUAD remains unclear. In this study, we investigated the intricate interplay of nesting apoptotic factors in LUAD. By analyzing nine key nesting apoptotic factors, we categorized LUAD patients into two distinct clusters. Further examination of immune cell profiles revealed that Cluster A exhibited greater infiltration of innate immune cells than did Cluster B. Additionally, we identified two genes closely associated with prognosis and developed a predictive model to differentiate patients based on molecular clusters. Our findings suggest that the loss of specific anoikis-related genes could significantly influence the prognosis, tumor microenvironment, and clinical features of LUAD patients. Furthermore, we validated the expression and functional roles of two pivotal prognostic genes, solute carrier family 2 member 1 (SLC2A1) and sphingosine kinase 1 (SPHK1), in regulating tumor cell viability, migration, apoptosis, and anoikis. These results offer valuable insights for future mechanistic investigations. In conclusion, this study provides new avenues for advancing our understanding of LUAD, improving prognostic assessments, and developing more effective immunotherapy strategies.

SPINK5 inhibits esophageal squamous cell carcinoma metastasis via immune activity.

BACKGROUND: Esophageal squamous cell carcinoma (ESCC) is a predominant subtype of esophageal cancer with relatively high mortality worldwide. Serine peptidase inhibitor Kazal-type 5 (SPINK5) is reported to be downregulated in ESCC. However, its explicit role in ESCC remains further investigation. METHODS: The tumor tissues and adjacent non-cancerous tissues were obtained from 196 patients with ESCC for the determination of SPINK5 mRNA levels. Additionally, the relationship between SPINK5 mRNA levels and clinicopathological features of ESCC patients was explored. The effects of SPINK5 on the invasion and migration of ESCC cells were assessed using Transwell assays. Furthermore, SPINK5 mRNA and LEKTI protein were measured in ESCC cell lines after treatment with poly (I:C), lipopolysaccharide (LPS) or unmethylated CpG DNA. Moreover, the correlation between expression of SPINK5 and nuclear factor-kappa B (NF-κB) signaling pathway-related genes was analyzed in the TCGA-ESCC cohort, and the effects of SPINK5 on NF-κB transcription was analyzed using a luciferase reporter gene assay. Finally, the correlations between SPINK5 and infiltration of immune cells, immune scores, stromal scores and ESTIMATE (i.e., Estimation of STromal and Immune cells in MAlignant Tumor tissues using Expression data) scores were explored. RESULTS: SPINK5 mRNA levels were downregulated in tumor tissues, which was significantly correlated with higher lymph node metastases. Overexpressed SPINK5 inhibited cell invasion and migration in ESCC cell lines. Mechanistically, LPS-induced activation of Toll-like receptor 4 (TLR4) decreased SPINK5 mRNA and LEKTI in KYSE150 and KYSE70 cells. Spearman correlation analysis revealed that SPINK5 mRNA was significantly negatively correlated with a total of seven NF-κB signaling pathway-related genes in TCGA-ESCC patients. Moreover, downregulation of SPINK5 increased and upregulation of SPINK5 decreased the activity of the NF-κB promoter in HEK293T cells. Finally, immune cells infiltration analysis revealed that SPINK5 was significantly correlated with the infiltration of various immune cells, stromal scores, immune scores and ESTIMATE scores. CONCLUSIONS: SPINK5 plays critical roles in the TLR4/NF-κB pathway and immune cells infiltration, which might contribute to the ESCC metastasis. The findings of the present study may provide a promising biomarker for the diagnosis and treatment of esophageal squamous cell carcinoma.

Ectopic Expression of TIGIT in Lung Adenocarcinoma and Its Clinical Significance.

This study aimed to investigate the T cell immunoreceptor with ITIM and Ig domains (TIGIT) expression in lung adenocarcinoma (LUAD). TIGIT expression was measured by western blot, reverse transcription quantitative polymerase chain reaction. Seventy-two paired surgical specimens were collected from patients with stage I-IV LUAD. The expression of TIGIT in surgical specimens was determined using immunohistochemistry. TIGIT was overexpressed in LUAD tissues. Moreover, overexpressed TIGIT was significantly associated with advanced clinical staging, lymph node metastasis, distant metastasis, and TP53 mutations in LUAD. Moreover, high expression of TIGIT was negatively correlated with purity of CD4+ T cells. High rations of TIGIT+CD4+ T cells predicted poor overall survival of LUAD patients. Additionally, high ratios of TIGIT+CD4+ T cells is closely related to CD4+ T cell depletion. Taken together, TIGIT was overexpressed in LUAD patients. High levels of TIGIT induced the alteration of CD4+ T cell based immunomodulation and predicted poor prognosis of LUAD patients. Therefore, TIGIT can be potential biomarker for LUAD.

The E3 ubiquitin ligase RNF135 modulates chemotherapy resistance to oxaliplatin for colorectal cancer by modulating autophagy.

BACKGROUND: RING finger protein 135 plays an important role in tumorigenesis and is associated with drug resistance. METHODS: Bioinformatics analysis showed that RNF135 was significantly differentially expressed in colorectal cancer. RT-qPCR and western blot were used to detect the expression of RNF135. Immunohistochemical analysis were used to measure the expression of RNF135 and Ki-67. RESULTS: The expression of RNF135 was up-regulated in human tissue samples and colorectal cancer and was positively correlated with Ki-67. Compared with oxaliplatin sensitive patients, RNF135 expression levels were higher in the tissue of resistant patients. The regulatory effect of RNF135 on colorectal cancer cells was further investigated in vitro. Therefore, inhibition of autophagy by down-regulating RNF135 can partially increase its susceptibility to oxaliplatin.

Whole ß-glucan particle attenuates AOM/DSS-induced colorectal tumorigenesis in mice via inhibition of intestinal inflammation.

Yeast ß-glucan is a polysaccharide purified from the Saccharomyces cerevisiae cell wall, and its multiple biological activities are essential for immune regulation. However, the effect of ß-glucan on the intestinal immune response during colitis-associated colorectal cancer (CAC) is unclear. Here, we explore the possible role of ß-glucan in the development of CAC. Wild type (WT) mice with CAC induced by azoxmethane (AOM) and dextran sodium sulfate (DSS) had fewer tumors than untreated mice after oral ß-glucan because of increased antitumor dendritic cells (DCs) in the tumor microenvironment, resulting in more CD8+ T cells and the production of related cytokines. ß-glucan also increased resistance to DSS-induced chronic colitis by reshaping the inflammatory microenvironment. These data suggest that ß-glucan improves experimental intestinal inflammation and delays the development of CAC. Therefore, ß-glucan is feasible for treating chronic colitis and CAC in clinical practice.

The use of the mean computed-tomography value to predict the invasiveness of ground-glass nodules: A meta-analysis.

The invasiveness of ground-glass nodules (GGNs) is difficult to characterize through morphological examination. Multiple studies have independently detected a close relationship between mean computed tomography value and invasiveness of GGNs, however, their relative diagnostic accuracy is uncertain. Here, we performed a meta-analysis to validate whether the mean computed tomography value can predict the invasiveness of GGNs. Briefly, we searched the Web of Science, Embase, PubMed, Cochrane, Google Scholar, CNKI, VIP, Wanfang and SinoMed databases. The sensitivity, specificity, 95% confidence interval (CI), symmetric receiver operating characteristic curve (SROC curve) and the area under curve (AUC) were obtained using STATA 16.0 to evaluate the predictive value of the mean computed tomography value for GGNs. The presence of heterogeneity was assessed using fixed effects sensitivity analysis and I2 statistics. We used the Deek's funnel plot to evaluate the possibility of publication bias. Thirteen studies encompassing 1564 GGNs were included in our meta-analysis. Six of these studies revealed that using the mean computed tomography value for the diagnosis of pre-invasive and invasive lesions had a sensitivity and specificity of 0.75 (95% CI: 0.61-0.85) and 0.81 (95% CI: 0.74-0.86), respectively. The optimal critical value was -557 Hu. Later, eight studies were examined for the use of the mean CT value for patients with minimally invasive adenocarcinoma (MIA) and invasive adenocarcinoma (IAC); the results showed that the sensitivity was 0.78 (95% CI: 0.66-0.86) and the specificity was 0.81 (95% CI: 0.68-0.89), and the optimal critical value was -484 Hu. Therefore, the mean computed tomography value assessed via CT scan could be a significant predictor of the invasiveness of GGNs as well as a good surgical treatment guide in patients diagnosed with lung cancer. PROSPERO REGISTRATION NUMBER: CRD42020177125.

Seasonal variations, temperature dependence, and sources of size-resolved PM components in Nanjing, east China.

Size-segregated ambient particulate matter (PM) samples were collected seasonally in suburban Nanjing of east China from 2016 to 2017 and chemically speciated. In both fine (< 2.1 µm, PM2.1) and coarse (> 2.1 µm, PM>2.1) PM, organic carbon (OC) accounted for the highest fractions (26.9% ± 10.9% and 23.1% ± 9.35%) of all measured species, and NO3- lead in average concentrations of water-soluble inorganic ions (WSIIs). The size distributions of measured components were parameterized using geometric mean diameter (GMD). GMD values of NO3-, Cl-, OC, and PM for the whole size range varied from < 2.1 µm in winter to > 2.1 µm in warm seasons, which was due to the fact that the size distributions of semi-volatile components (e.g., NH4NO3, NH4Cl, and OC) had a dependency on the ambient temperature. Unlike OC, elemental carbon (EC), and elements, NH4+, NO3-, and SO42- exhibited an increase trend in GMD values with relative humidity, indicating that the hygroscopic growth might also play a role in driving seasonal changes of PM size distributions. Positive matrix factorization was performed using compositional data of fine and coarse particles, respectively. The secondary formation of inorganic salts contributing to the majority (> 70%) of fine PM and 20.2% ± 19.9% of speciated coarse PM. The remaining coarse PM content was attributed to a variety of dust sources. Considering that coarse and fine PM had comparable mass concentrations, more attention should be paid to local dust emissions in future air quality plans.

Differential expression of Tim3 protein in colorectal cancer associated with MSI and Braf mutation.

Tim3 is a negative immune checkpoint molecule and plays a crucial part in tumor-induced immune suppression. Tim3 is a cell surface molecule expressed on T cells marking dysfunctional CD8+ cells in various kinds of cancers. Tim3 expression was mainly reported in tumor-infiltrating lymphocytes (TILs). There are few studies focusing on the expression of Tim3 in tumor cells. Immunohistochemistry was performed to determine Tim3 expression level. The relationships between Tim3 expression in colorectal cancer cells and in tumor-infiltrating lymphocytes and cilicopathological parameters were statistically analyzed. Tim3 was differentially detected in TILs and in colorectal cancer cells. Positive expression of Tim3 in colorectal cancer cells was associated with tumor location (P=0.001), depth of tumor invasion (P<0.001), lymph node metastasis (P=0.001), TNM stage (P=0.001), MSI (P=0.008), and Braf V600E mutation (P=0.001). On the other hand, positive expression of Tim3 in TILs was only related to depth of tumor invasion (P<0.001). Positive expression of Tim3 in both colorectal cancer cells and TILs was associated with depth of tumor invasion (P<0.001), lymph node metastasis (P=0.002), TNM stage (P=0.002), MSI (P=0.039), and Braf V600E mutation (P=0.009). Kaplan-Meier survival analysis showed that Tim3 expression in colorectal cancer and in TILs was significantly associated with patient overall survival (OS) rate (P=0.039, and 0.001). Tim3 may be a potential prognostic marker and a therapy target for colorectal cancer.

Efficacy of afatinib in a patient with rare EGFR (G724S/R776H) mutations and amplification in lung adenocarcinoma: A case report.

BACKGROUND: The most common EGFR mutations are in-frame deletions in exon 19 and point mutations in exon 21. Cases with classical EGFR mutations show a good response to EGFR tyrosine kinase inhibitors (TKIs), the standard first-line treatment. With the development of next generation sequencing, some uncommon genomic mutations have been detected. However, the effect of TKIs on such uncommon EGFR mutations remains unclear. CASE SUMMARY: Here, we report a case of rare EGFR co-mutation in non-small cell lung cancer and the efficacy of afatinib on this EGFR co-mutation. A 64-year-old woman was diagnosed with thoracolumbar and bilateral local rib bone metastases, bilateral pulmonary nodules, and pericardial and left pleural effusion. The pathological diagnosis was lung adenocarcinoma. To seek potential therapeutic regimens, rare co-mutation comprising rare EGFR G724S/R776H mutations and amplification were identified. The patient experienced a significant clinical response with a progression-free survival of 17 mo. CONCLUSION: A case of non-small cell lung cancer with rare EGFR G724S/R776H mutations and EGFR amplification responds well to TKI treatment.

NLK interacts with 14­3­3ζ to restore the expression of E­cadherin.

The Nemo­like kinase (NLK), a conserved serine/threonine kinase, plays a critical role in the regulation of a variety of transcription factors, with important roles in determining cell fate. Although recent studies have demonstrated decreased expression patterns of NLK in various types of human cancer, the functional mechanism of NLK in cancer development has not been elucidated. Here, in the present study overexpression of NLK was found to inhibit the growth and migration of the non­small cell lung cancer A549 cell line. NLK was subsequently found to interact with 14­3­3ζ (also known as YWHAZ), which is responsible for E­cadherin silencing during epithelial­mesenchymal transition (EMT). Furthermore, NLK overexpression was able to restore the expression of E­cadherin inhibited by 14­3­3ζ. Notably, NLK interacts with 14­3­3ζ and prevents its dimerization, which is essential for 14­3­3ζ stability and function. By fusing two copies of the 14­3­3ζ gene, via a Gly­rich linker, a non­dissociable dimer of 14­3­3ζ was formed. It was found that NLK was unable to restore the expression of E­cadherin inhibited by the overexpression of the fused dimer of 14­3­3ζ. In addition, the increased ability of migration induced by the overexpression of fused 14­3­3ζ dimer could not be altered by NLK overexpression. The results from the present study indicate that NLK is a negative regulator of 14­3­3ζ and plays a tumor suppressive role in the inhibition of cancer cell migration.

PM2.5 elements at an urban site in Yangtze River Delta, China: High time-resolved measurement and the application in source apportionment.

Elemental concentrations of ambient aerosols are commonly sampled over 12-24 h, and the low time resolution puts a great limit on current understanding about the temporal variations and source apportionment based on receptor models. In this work, hourly-resolved concentrations of eighteen elements in PM2.5 at an urban site in Nanjing, a megacity in Yangtze River Delta of east China, were obtained by using a Xact 625 ambient metals monitor from 12/12/2016 to 12/31/2017. The influence of traffic activities was clearly reflected by the spikes of crustal elements (e.g., Fe, Ca, and Si) in the morning rush hour, and the firework burning and sandstorm events during the sampling periods were tracked by sharp enrichment of Ba, K and Fe, Ca, Si, Ti in PM2.5, respectively. To evaluate the advantage of hourly-resolved elements data in identifying impacts from specific emission sources, positive matrix factorization (PMF) analysis was performed with the 1-h data set (PMF1-h) and 23-h averaged data (PMF23-h), respectively. The 4- and 6-factor PMF23-h solutions had similar factor profiles and consistent factor contributions as the corresponding PMF1-h solutions. However, due to the limit in inter-sample variability, PMF analysis with 23-h average data misclassified some major (e.g., K, Fe, Zn, Ca, and Si) and trace (e.g., Pb) elements in factor profiles, resulting in different absolute factor contributions between PMF23-h and PMF1-h solutions. These results suggested the use of high time-resolved data to obtain valid and robust source apportionment results.

Characteristics of heterotrophic nitrification and aerobic denitrification bacterium Acinetobacter sp. T1 and its application for pig farm wastewater treatment.

A novel heterotrophic bacterium was isolated from activated sludge of a pig farm wastewater treatment plant and identified as Acinetobacter sp. T1. It exhibited efficient heterotrophic nitrification and aerobic denitrification capability to utilize ammonium, nitrate or nitrite as the sole nitrogen source, and their removal rates were 12.08, 5.53 and 1.69 mg/L/h, respectively. Furthermore, the optimal conditions for the heterotrophic nitrification process were sodium citrate as the carbon source, C/N mass ratio of 10, pH of 8.5 and dissolved oxygen (DO) concentration of 5.1 mg/L. Only trace amounts of nitrate and nitrite were observed during the process. When the aerobic tank of the A2O process of a pig farm wastewater treatment plant was inoculated with traditional activated sludge, the average removals of COD, NH4+- N and TN in the effluent were 30%, 15% and 16%, respectively, which was much lower than that of inoculated with strain T1, the increase was statistically significant, indicating a great potential of strain T1 for full-scale applications.