Correlation between chromatin epigenetic-related lncRNA signature (CELncSig) and prognosis, immune microenvironment, and immunotherapy in non-small cell lung cancer.

Chromatin regulators drive cancer epigenetic changes, and lncRNA can play an important role in epigenetic changes as chromatin regulators. We used univariate Cox, LASSO, and multivariate Cox regression analysis to select epigenetic-associated lncRNA signatures. Twenty-five epigenetic-associated lncRNA signatures (CELncSig) were identified to establish the immune prognostic model. According to Kaplan-Meier analysis, the overall survival of the high-risk group was significantly lower than the low-risk group. Receiver operating characteristic (ROC) curves, C-index, survival curve, nomogram, and principal component analysis (PCA) were performed to validate the risk model. In GO/KEGG analysis, differentially expressed lncRNAs were correlated with the PI3K-Akt pathway, suggesting that they were highly associated with the metastasis of LUAD. Interestingly, in the immune escape analysis, the TIDE score was lower, and the possibility of immune dysfunction is also slighter in the high-risk group, which means they still have the potential to receive immunotherapy. And CELncsig is highly correlated with immune pathways T_cell_co-inhibition and Check-point. Also, the IMvigor210 cohort analysis indicated that our risk-scoring model has significant potential clinical application value in lung cancer immunotherapy. And we also screened out ten potential chemotherapy agents using the 'pRRophetic' package.

Comprehensive transcriptomic and co-expression analysis of ABL1 gene and molecularly targeted drugs in hepatocellular carcinoma based on multi-database mining.

Hepatocellular carcinoma (HCC) is one of the leading causes of cancer death worldwide. Consequently, it is essential to identify biomarkers for treatment response and the prognosis prediction. We investigated whether ABL1 can function as a biomarker or a drug target for HCC. We assessed the ABL1 expression, genetic alterations and patients' survival from LinkedOmics, GEO, TCGA and Human Protein Atlas. We analyzed PPI, GO and KEGG pathways. GSEA was analyzed for functional comparison. The current drugs targeting ABL1 were statistically analyzed using DRUGSURV and DGIdb database. We found ABL1 is overexpressed in HCC and its higher expression reduces survival probability. Genetic changes of ABL1 are not frequent. We screened out 25 differentially expressed genes correlated with ABL1. The top functions of ABL1 are biological regulation, metabolic process, protein-containing, and protein binding. KEGG pathways showed that ABL1 and correlated with ABL1 significantly genes markedly enriched in the ErbB signaling pathway, and pathways in cancer. We counted the existing drugs targeting ABL1, which indicates that inhibiting ABL1 expression may improve the survival probability of HCC. In conclusion, ABL1 plays a crucial role in the development and progression of this cancerization and is a potential drug target.

Correlation Analysis Among the Level of IL-35, Microvessel Density, Lymphatic Vessel Density, and Prognosis in Non-Small Cell Lung Cancer.

The aim of this study was to determine the expression of IL-35 and the lymphatic vessel density (LVD) and microvessel density (MVD) in the pathological tissues from patients with non-small cell lung cancer (NSCLC) and to analyze their correlation with other common clinical prognostic factors, as well as patients' overall survival and progression-free survival. We analyzed the pathological characteristics of 130 patients with NSCLC and determined the IL-35 expression, MVD, and LVD changes in the pathological tissues by immunohistochemistry. The results showed that IL-35 expression was significantly correlated with tumor differentiation, lymph node metastasis, T staging, LVD, and MVD (P < 0.05) but was not associated with age, sex, smoking, and other factors. Univariate analysis of risk models showed that age, lymph node metastasis, T stage, and high IL-35 expression, LVD, and MVD were significantly associated with NSCLC prognosis (P < 0.05), whereas sex, smoking, and high differentiation were not correlated with prognosis. Multivariate analysis of the proportional risk model showed that the IL-35 expression, lymph node metastasis, high LVD, and high MVD were significantly correlated with NSCLC prognosis (P < 0.05). In conclusion, IL-35, MVD, and LVD may be independent prognostic markers. In addition, IL-35 might represent a promising clinical drug target for the treatment of NSCLC.

Temporal Variations of Water Chemistry in the Wet Season in a Typical Urban Karst Groundwater System in Southwest China.

It is important to investigate temporal variations of water chemistry for the purpose of improving water quality in karst groundwater systems. Groundwater samples were collected daily under various land uses of Guiyang. Major ions and stable carbon isotope composition of dissolved inorganic carbon (δ13CDIC) were analyzed to understand the biogeochemical processes. The water chemistry was dominated by Ca2+, Mg2+, HCO3-, and SO42-, which mainly derived from the dissolution of carbonate rocks (limestone and dolomite) and oxidation of sulfide. The groundwater was defined as of the HCO3-Ca Mg and HCO3·SO4-Ca·Mg type, according to its hydrochemical characteristics. Results suggested that hydrochemical concentrations changed quickly, in response to rainfall events. The fast response revealed that karst groundwater was easily impacted by rainfall and anthropogenic inputs according to temporal variation of water chemistry. The distribution of DIC (dissolved inorganic carbon) and δ13CDIC showed that DIC is mainly sourced from soil CO2(g) influx and carbonate dissolution. δ13CDIC and major ions ratios suggested that carbonate minerals were dissolved by H2SO4 at groundwater in wooded area, contributing an important source for DIC due to the slight enrichment of heavy δ13CDIC. More negative δ13CDIC values were observed after rainfall reflected the fact that soil CO2(g) and organic carbon oxidation influxes accounted for a large share during DIC formation. Various δ13CDIC and hydrochemical patterns were observed under various land use and human activity conditions. Meanwhile, relative high nitrate loads were found in groundwater after rainfall, suggesting high anthropogenic inputs following rainwater as having side effects on water quality. This study suggests that water chemistry and isotopic proof provide a better understanding of water quality and carbon dynamics responding to rainfall events in the karst groundwater systems.

Reciprocal regulation of miR-206 and IL-6/STAT3 pathway mediates IL6-induced gefitinib resistance in EGFR-mutant lung cancer cells.

Persistently activated IL-6/STAT3 pathway promotes acquired resistance to targeted therapy with epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) in non-small-cell lung cancer (NSCLC) treatment. miR-206 has been verified to be dysregulated and plays as a negative regulator in lung cancer. However, whether miR-206 may overcome IL6-induced gefitinib resistance in EGFR-mutant lung cancer remains elusive. In this study, we investigated the role of miR-206 in IL6-induced gefitinib-resistant EGFR-mutated lung cancer cell lines. We showed that forced miR-206 expression restored gefitinib sensitivity in IL6-induced gefitinib-resistant EGFR-mutant lung cancer cells by inhibiting IL6/JAK1/STAT3 pathway. Specifically, mechanistic investigations revealed that miR-206 blocked IL-6/STAT3 signalling via directly targeting the 3'-UTR of intracellular IL-6 messenger RNA. Moreover, IL-6 induced miR-206 down-regulation by reducing the cropping process of primary miR-206 (pri-miR-206) into the Drosha/DGCR8 complex. Taken together, our findings reveal a direct role of miR-206 in regulating IL-6/STAT3 pathway and contrarily activated IL-6/STAT3 signalling mediates the miR-206 maturation process in gefitinib-resistant EGFR-mutant lung cancer cells.

Clinical significance of FAP-α on microvessel and lymphatic vessel density in lung squamous cell carcinoma.

AIMS: We aimed to determine whether cancer-associated fibroblasts (CAFs) are associated with microvessel density (MVD) and lymphatic vessel density (LVD) in lung squamous cell carcinoma, as well as their clinical significance in predicting survival. METHODS: 122 patients were enrolled in the study. Samples were obtained on resection at the Department of Thoracic Surgery of the Qingdao Municipal Hospital between January 2011 and December 2014. Immunohistochemistry was used to determine vessel and lymphatic vessel density, and CAF abundance (fibroblast activation protein α (FAP-α) positivity). Statistical analyses were performed on 85 patients to test for correlation of CAF density and other clinicopathological variables with 3-year overall survival (OS) and disease-free survival (DFS). RESULTS: High stromal CAF abundance significantly correlated with increased MVD and LVD in lung squamous cell carcinoma (p<0.05). χ2 test revealed a significant association of CAF density with lymph node metastasis. Cox proportional hazards model showed that both higher CAF density and lymph node metastasis negatively correlate with survival. CAF density or lymph node status can be used as an independent prognostic factor to predict 3-year OS and DFS. CONCLUSIONS: CAF density, identified by FAP-α staining pattern, should be considered as a novel biomarker for disease prognosis in patients with lung squamous cell carcinoma.

[Expression of miR-34a in Bone Marrow of Adult Acute Lymphoblastic Leukemia and Its Significance in Cell Drug Resistance].

OBJECTIVE: To investigate the expressions of miR-34a in bone marrow of adult acute lymphoblastic leukemia (ALL) and its relationship with drug resistance. METHODS: Forty-seven cases of newly diagnosed adult ALL were selected and their bone marrow samples were taken at the time of newly diagnosed and relapsed or complete remission; 26 pairs of specimens were in newly diagnosed-complete remission group, and 21 pairs of specimens were in newly diagnosed-relapse group. The expressions of miR-34a in bone marrow samples, CCRF-CEM cells and resistant CEM-C1 cell strains were detected by real-time quantitative PCR. The expression of miR-34a in CCRF-CEM cells was inhibited and was increased in CEM-C1 cells detected by electroporation transfection method. All the cells were incubated at different concentration of camptothecin. The cell survival was analyzed by CCK-8 method, the cell proliferation inhibition rate (%) and resistance index (RI) were calculated. RESULTS: In newly diagnosed-complete remission group, the miR-34a expression at newly diagnosis was significantly lower than that in complete remission and the control group, the differences were statistically significant (P < 0.05). In newly diagnosed-relapsed group, the miR-34a expressions at newly diagnosis and relapse were lower than those in the control group, the differences were statistically significant (P < 0.05). The expression level in CEM-C1 cells was (2.64 ± 1.37) which significantly lower than that in CCRF-CEM cells (5.14 ± 2.06), the differences were statistically significant (P < 0.05). The expression level of miR-34a in CCRF-CEM cells transfected with miR-34a inhibitor was (3.14 ± 1.15), which significantly lower than that in the miRNA inhibitor-negative control group, the difference was statistically significant (P < 0.05). The cell proliferation inhibition rate of CCRF-CEM cells transfected with miR-34a inhibitor was significantly higher than that in the negative control transfectedcells (P < 0.05), the IC(50) was 28.73 ng/mL and 2167.00 ng/mL respectively, and RI = 75.43. The expression level of miR-34a in CEM-C1 cells transfected with miR-34a mimic was (5.06 ± 1.73), which was significantly higher than that in the miRNA mimics transteted-negative control CEM-C1 cells (P < 0.05). The proliferation inhibition rate in CEM-C1 cells transfected with miR-34a mimic was significantly lower than that in the negative control-transfected cells (P < 0.05). The IC(50) was 112.57 ng/mL and 1.27 ng/mL respectively, and the RI = 88.64. CONCLUSION: The expression of miR-34a in bone marrow samples of adult ALL is low which may be associated with the relapse and drug resistance of ALL.

Co-expression of pregnane X receptor and ATP-binding cassette sub-family B member 1 in peripheral blood: A prospective indicator for drug resistance prediction in non-small cell lung cancer.

The aim of the present study was to investigate the protein expression profiling of pregnane X receptor (PXR) and ATP-binding cassette sub-family B member 1 (ABCB1; also known as MDR1 or P-gp), present in the peripheral blood mononuclear cells (PBMCs) and cancerous tissues of cases of non-small cell lung cancer (NSCLC). Furthermore, the study aimed to assess the feasibility of predicting drug resistance through the medium of PBMCs. Of the subjects included in the study, 37 were histopathologically diagnosed with NSCLC and 17 were control patients without cancer. ThinPrep liquid-based smears with cytosine were applied in the examination of the PBMCs and proved quite effective in preserving the morphology and surface antigens of the lymphocytes. Measurements of expression levels in the PBMCs and cancerous tissues were obtained by immunohistochemical means. The results showed that, with the exception of the selective PXR expression in the normal lung tissues, the two types of proteins existed extensively throughout the PBMCs, normal tissues and tumors. Among the cancer patients, prior to chemotherapy, a significant rise in ABCB1 expression could be observed in the PBMCs, together with a similar rise in ABCB1 and PXR expression in the tumor specimens. Marked upregulation of the two proteins was detected in the PBMCs following 1 cycle of first-line chemotherapy. ABCB1 expression, correlated with PXR, persisted mostly in the PBMCs and tissue samples. When bound to and activated by ligands, PXR translocates from the cytoplasm to the nucleus of the cells. PXR subsequently binds to its DNA response elements as a heterodimer with the retinoid X receptor. A PXR translocation of moderate or low differentiation was identified in 3 cases of adenocarcinoma, which were co-expressing the two genes in the PBMCs prior to chemotherapy. During follow-up visits, tumor recurrence was observed within 3 months in 5 cases, which were characterized by PXR translocation. These findings indicate that the combined expression of PXR and ABCB1 in PBMCs may be used as a prospective indicator in diagnosis prior to histopathological diagnosis, and therefore may function as a novel biomarker for the prediction of drug resistance.

Increased expression of apoptosis-related protein 3 is highly associated with tumorigenesis and progression of cervical squamous cell carcinoma.

The apoptosis-related protein 3 (APR3) gene was first cloned from HL-60 cells treated with all-trans-retinoic acid and was thought to be related to tumor cell apoptosis or differentiation. In this study, we sought to investigate its expression profile in cervical squamous cell carcinoma (SCC) and preneoplastic lesions to determine whether APR3 is involved in the malignant progression of SCC. The purified partial recombinant APR3 proteins were used to immunize rabbits for raising antibodies, and the specificity of the polyclonal anti-APR3 antibody was determined by enzyme-linked immunosorbent assay and Western blot. Sections were assessed for APR3 expression by immunohistochemistry in archived tissues from human normal cervix samples (n = 20), cervical intraepithelial neoplasia (n = 19), and invasive SCC (n = 52). Specific cytoplasmic immunostaining was evaluated for overall intensity and uniformity to derive a combined histoscore. The results of enzyme-linked immunosorbent assay and Western blot indicated that anti-APR3 antibody can serve as a good tool for research. The immunohistochemical analysis demonstrated an increased expression of APR3 in SCC relative to normal cervix epithelium and cervical intraepithelial neoplasia (P < .05). Strikingly, APR3 expression level was significantly higher in nonkeratinizing SCCs compared with keratinizing SCCs (P < .05) and higher in carcinomas with lymph node metastasis compared with cases without lymph node metastasis (P < .05). This study demonstrates that APR3 expression is increased significantly with malignant progression of human cervical SCC, and thus, it may serve as a potential biomarker to predict prognosis of cervical SCC.

[Chromophobe renal cell carcinoma: a clinicopathologic study and immunophenotypes of 42 cases].

OBJECTIVE: To study the clinicopathologic features, immunohistochemical profiles and prognosis of chromophobe renal cell carcinoma (ChRCC). METHODS: Forty-two cases of ChRCC were retrieved from the archival files of the Affiliated Hospital of Qingdao University, 401 Hospital of PLA and Qingdao Municipal Hospital from 2003 to 2011. The clinical and pathologic features of the tumors were reviewed. Hale colloidal iron staining was performed and EnVision immunohistochemistry was used to detect the expression of a series of immunologic markers. Forty cases of clear cell renal cell carcinoma and 10 cases of renal oncocytoma were selected as controls. RESULTS: The patients included 17 males and 25 females. The age of patients ranged from 39 years to 78 years (median age = 57 years). On gross examination, the tumors ranged from 2 cm to 19 cm in greatest dimension (mean size = 7.3 cm). Histologically, the tumors were mainly composed of solid sheets, acini or tubules of malignant cells. The tumor cells contained clear finely reticular ("chromophobe") and eosinophilic cytoplasm with perinuclear clearing. The nuclear outline was irregular and wrinkled. Nucleoli were inconspicuous and mitotic figures were barely seen. Hale colloidal iron stain was positive in all cases. Immunohistochemically, the tumor cells were variably positive for EMA (100%, 42/42), CK7 (95.2%, 40/42), Ksp-cad (92.9%, 39/42), CK18 (88.1%, 37/42), CD117 (61.9, 26/42), CD10 (31.0%, 13/42) and PAX2 (28.6%, 12/42). They were negative for vimentin, CA IX and TFE3. The follow-up period in 31 patients ranged from 2 to 77 months (average duration = 29 months). Three patients died of tumor metastasis 3, 8, 13 months respectively after the operation. Twenty-eight patients were still alive without evidence of tumor recurrence. CONCLUSIONS: ChRCC predominantly occurs in middle-aged and elderly patients. It often carries a favorable prognosis. The presence of plant cell-like morphology, pale cells with uniform reticular microvesicular appearance and perinuclear clearing are characteristic histologic features. The diffuse positivity for Hale colloidal iron stain and EMA/CK7/Ksp-cadherin/CD117-positive immunoprofiles are also useful in differential diagnosis.