Identification of new biomarkers in immune microenvironment of testicular germ cell tumour.

To seek novel prognostic biomarkers for testicular germ cell tumour (TGCT) and investigate the tumour immune microenvironment, we identified critical differentially expressed genes (DEGs) by overlapping GSE1818 dataset from Gene Expression Omnibus (GEO). Protein-protein interaction (PPI) network was used to investigate key modules and hub genes. Functional enrichment analysis was performed to investigate the underlying molecular functions of the DEGs in TGCT development and progression. The following survival analysis based on The Cancer Genome Atlas (TCGA) TGCT dataset indicated that AKAP4, SPA17 and TNP1 are correlated with TGCT prognosis. Immunohistochemistry and quantitative real-time polymerase chain reaction verified the down-regulation of the 3 hub genes in TGCT. Gene set enrichment analysis was conducted to further explore the role of the 3 hub genes in TGCT respectively. In addition, TGCT samples had high infiltration of CD8+ T cells, M0 and M1 macrophage cells, and resting myeloid dendritic cells in immune microenvironment. We also constructed the microRNA-gene regulatory networks to identify the key upstream microRNAs in TGCT. In conclusion, our findings indicated that AKAP4, SPA17 and TNP1 are promising biomarkers of TGCT. AKAP4 and TNP1 might regulate immune cells infiltration in immune microenvironment.

Evaluating the biological functions of the prognostic genes identified by the Pathology Atlas in bladder cancer.

The prognosis­associated genes of urinary bladder cancer have been systematically investigated in the Pathology Atlas project based on The Cancer Genome Atlas data. However, the biological functions of most genes in bladder cancer remain unknown. The present study investigated the biological function of 12 of the most significant survival­associated genes (ABRACL, MITD1, ZNF524, EMP1, HSPB6, CXorf38, TRIM38, ZNF182, ZNF195, SPRN, PTPN6 and LIPT1) in urothelial cancer reported by the Pathology Atlas project, with respect to cell proliferation and migration. In vitro, proliferation and migration analyses of T24 cells were performed following the transfection of the 12 prognostic genes. The results were validated with a small interfering (si)RNA library. Immunohistochemistry (IHC) analysis of clinical samples was performed to determine the association between gene expression and tumor metastasis. Furthermore, RNA sequencing was used to investigate the downstream signals. Among the 12 prognostic genes, MIT­domain containing protein 1 (MITD1) transfection was demonstrated to inhibit T24 cell migration to a certain degree. Experiments performed with a 7­gene siRNA library demonstrated that MITD1 knockdown markedly upregulated cell migratory abilities. Mechanistically, the influence of MITD1 on cell signal transduction was assessed via RNA sequencing. Cell migration­associated genes, including KISS1, SPANXB1, SPINT1, PIWIL2, SNAI1, APLN and CTHRC1 were dysregulated. IHC analysis demonstrated that MITD1 protein expression was notably lower in metastatic lymph nodes compared with the primary tumors. Taken together, the results of the present study suggest that the prognostic gene, MITD1 may serve as a migration inhibitor, and be developed as a potential therapeutic target for improving the prognosis of bladder cancer.

N-acetyltransferase 2 polymorphism is associated with bladder cancer risk: An updated meta-analysis based on 54 case-control studies.

OBJECTIVE: N-acetyltransferase 2 (NAT2) polymorphism could participate in the metabolism of carcinogens through regulating the activity of a series of critical enzymes. However, the effects of NAT2 polymorphism on bladder cancer (BCa) risk were still inconclusive. In order to illustrate whether NAT2 polymorphism may influence the susceptibility to BCa, we conducted this updated meta-analysis. MATERIALS AND METHODS: Databases including PubMed, Medline, Embase, Web of Science, Cochrane Library, and China National Knowledge Infrastructure(CNKI) were systematically retrieved and we applied MetaGenyo to perform final meta-analysis. Odds ratios (ORs) as well as 95% confidence intervals (CIs) were calculated and Bonferroni method was applied to correct the P-value for multiple comparisons. The registration of this study protocol is at PROSPERO and ID is CRD42019133957. RESULTS: Ultimately, 54 case-control studies were identified for final meta-analysis (13343 BCa cases and 18,586 controls). Overall analysis indicated that the slow genotype in NAT2 polymorphism was obviously associated with BCa risk (PBonferroni < 0.001). Subgroup analyses demonstrated that significant risk with the slow genotype was observed in Caucasians, Asians, smokers, non-exposed individuals, high grade bladder cancer (HGBC) patients and muscle-invasive bladder cancer (MIBC) patients. In addition, the intermediate NAT2 genotype was revealed to increase the BCa risk of Asians and transitional cell carcinoma (TCC) patients. However, no correlation was identified in Africans with the NAT2 polymorphism. CONCLUSIONS: The slow NAT2 genotype was identified to be the risk genotype for BCa. The intermediate genotype could serve as the candidate risk genotype. The gene-smoking interaction with NAT2 polymorphism might accelerate the tumor progression.

Concurrent computed tomography-guided radioactive iodine-125 seeds percutaneous interstitial implantation and chemotherapy for treatment of cervical lymph node metastases.

AIM: The study aimed to evaluate the effect of concurrent computed tomography (CT)-guided percutaneous interstitial implantation of iodine-125 (125I) seeds and chemotherapy on cervical lymph nodes metastasis. METHODS: The prospective randomized study included 82 cases with cervical lymph nodes metastasis who were admitted to our hospital from January 2010 to June 2012. All the subjects were randomly divided into the concurrent 125I implantation and chemotherapy group (n = 48) and chemotherapy-only group (n = 34) according to the treatment strategy. The concurrent 125I implantation and chemotherapy group was treated with CT-guided 125I seeds implantation and routine chemotherapy. The routine chemotherapy included paclitaxel and cisplatin. Patients were followed up for 6 months. RESULTS: In the concurrent 125I implantation and chemotherapy group, overall response rate (complete response [CR] + partial response [PR]) was 82.61% and 85.51% at 2 and 6 months posttreatment, respectively. The longest diameter of CR and PR lymph nodes was markedly decreased after treatment (P < 0.05). In the chemotherapy-only group, overall response rate was 22.45% and 10.20% at 2 and 6 months posttreatment, respectively. The number of patients with moderate to severe pain was much less in concurrent 125I implantation and chemotherapy group than that of chemotherapy-only group (4.17% vs. 17.64%; P < 0.05) at 6-month posttreatment. No treatment-related death or severe complication was reported in the two groups. CONCLUSION: Concurrent CT-guided 125I seeds implantation and chemotherapy is superior to routine chemotherapy in efficacy, safety, and pain relief in patients with cervical lymph nodes metastasis.

The PI3K/AKT Pathway and Renal Cell Carcinoma.

The phosphatidylinositol 3 kinase (PI3K)/AKT pathway is genetically targeted in more pathway components and in more tumor types than any other growth factor signaling pathway, and thus is frequently activated as a cancer driver. More importantly, the PI3K/AKT pathway is composed of multiple bifurcating and converging kinase cascades, providing many potential targets for cancer therapy. Renal cell carcinoma (RCC) is a high-risk and high-mortality cancer that is notoriously resistant to traditional chemotherapies or radiotherapies. The PI3K/AKT pathway is modestly mutated but highly activated in RCC, representing a promising drug target. Indeed, PI3K pathway inhibitors of the rapalog family are approved for use in RCC. Recent large-scale integrated analyses of a large number of patients have provided a molecular basis for RCC, reiterating the critical role of the PI3K/AKT pathway in this cancer. In this review, we summarize the genetic alterations of the PI3K/AKT pathway in RCC as indicated in the latest large-scale genome sequencing data, as well as treatments for RCC that target the aberrant activated PI3K/AKT pathway.

The status, limitation and improvement of adoptive cellular immunotherapy in advanced urologic malignancies.

In recent years, immunotherapy has been gradually established as the fourth frequently adopted antitumor therapy, following surgery, chemotherapy and radiotherapy, for advanced urologic malignancies with an improved understanding of theoretical basis, such as molecular biology and immunology. Thereinto, adoptive cellular immunotherapy (ACI) has become one of the hotspots, which comprises a variety of treatment approaches, such as TIL, CIK cell, γδ T cell, CAR-engineered T cell and Allogeneic stem cell transplantation (alloSCT). Although preclinical efficacy has been demonstrated remarkably, clinical trials could not consistently show the benefit due to multi-factors in complex immunosuppressive microenvironment in vivo compared to that of in vitro. Here we review some timely aspects of ACI for advanced urologic malignancies, and describe the current status and limitation of immunotherapy from the cellular level. It's our expectation to provide prompting consideration of novel combinatorial ACI strategies and a resurgence of interest in ACI for advanced urologic malignancies.

Effect of anesthesia on cognitive status and MMP-2 expression in rats.

OBJECTIVE: To investigate the effect of anesthesia on the cognitive status damage and MMP-2 expression in rats. METHODS: A total of 120 healthy rats were selected and randomly divided into the control group, CF3-CH(OCH2F)-CF3 (Sevoflurane) group and CF3-CH2-O-CHF-CF3 group (Sevoflurane) (n=40). After training for 3 d by the Morris water maze, the control group were injected with fentanyl for analgesia, the CF3-CH(OCH2F)-CF3 group and the CF3-CH2-O-CHF-CF3 group were anesthesia with CF3-CH (OCH2F)-CF3 and CF3-CH2-O-CHF-CF3 on the basis of fentanyl, then rats in three groups underwent open surgery and suture conventional incision. Morris water maze was used to measure the rats' cognitive ability in three groups on the 1st d, 3rd d, 5th d and 7th d, and the brain tissue MMP-2 expression was detected. RESULTS: After 1 d/7 d of the surgery, Morris water maze performance and MMP-2 expression were not significantly different among three groups (P>0.05); After 3 d/5 d of the surgery, compared with the control group, the Morris water maze test result was significantly worsened, MMP-2 expression levels were significantly increased (P<0.05); After 3 d/5 d of the surgery, compared with the CF3-CH2-O-CHF-CF3 group, Morris water maze test result of CF3-CH(OCH2F)-CF3 group was significantly worsened, MMP-2 expression levels were significantly increased (P<0.05). CONCLUSIONS: Anesthesia can cause some injury on cognitive status, different anesthetic drugs may cause different injury, and the cognitive status injury is related to the MMP-2 expression.

Bioinformatics analysis of aggressive behavior of breast cancer via an integrated gene regulatory network.

BACKGROUND: Breast cancer is one of the most frequently diagnosed cancers in women. Though death from this disease is mainly caused by the metastases of the aggressive cancer cells, few studies have expounded the aggressive behavior of breast cancer. MATERIALS AND METHODS: We downloaded the gene expression profiles of GSE40057, including four aggressive and six less-aggressive breast cancer cell lines, from Gene Expression Omnibus and identified the differentially expressed genes (DEGs) between the aggressive and less-aggressive samples. An integrated gene regulatory network was built including DEGs, microRNAs (miRNAs), and transcription factors. Then, motifs and modules of the network were identified. Modules were further analyzed at a functional level using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway to study the aggressive behavior of breast cancer. RESULTS: A total of 764 DEGs were found and two modules were filtered from the integrated gene regulatory network. Totally two motifs and modules for DEGs were identified. Significant GO terms associated with cell proliferation and hormone stimulus of the modules were found and the target genes identified were  CAV1, CD44, and TGFßR2. The KEGG pathway analysis discovered that CAV1 and FN1 were significantly enriched in focal adhesion, extracellular matrix (ECM)-receptor interaction, and pathways in cancer. CONCLUSION: Aggressive behavior of breast cancer was proved to be related to cell proliferation and hormone stimulus. Genes such as CAV1, CD44, TGFßR2, and FN1 might be potential targets to diagnose the aggressive behavior of breast cancer cells.

Dose-volume histogram parameters for predicting radiation pneumonitis using receiver operating characteristic curve.

OBJECTIVE: To assess the predictability of dose-volume histogram (DVH) parameters for radiation pneumonitis (RP) using receiver operating characteristic (ROC) curve. METHODS: One hundred and thirty-five cases of locally advanced non-small cell lung cancer patients treated with three-dimensional radiotherapy and chemotherapy were analyzed retrospectively. The end point of follow-up was ≥2 grade RP defined according to the National Cancer Institute Common Terminology Criteria for Adverse Events, version 3.0. The ROC curve was used to explore the predictive sensitivity (SEN), specificity (SPE), positive predictive value (PPV), and negative predictive value (NPV) for potential DVH parameters associated with RP. RESULTS: Relative volumes of total lungs receiving ≥5 Gy (V(5)), ≥10 Gy (V(10)), ≥13 Gy (V(13)), ≥20 Gy (V(20)), and mean lung dose (MLD), were all correlated to the development of RP (p < 0.05), among which V 5 and V 20 were the most important factors (p = 0.045 and 0.037; OR = 3.166 and 3.030). However, collinearity was found between V(5) and V(20) (Spearman's rho 0.771, p < 0.01). The area under the ROC curve was 0.643 and 0.648 for using V(5) and V(20) as predictors. If predictive cut-off values were established as follows: V(5) = 0.8 and V(20) = 0.3, the parameters could provide predictive SEN, SPE, PPV and NPV were 0.387 and 0.581, 0.882 and 0.701, 0.444 and 0.321, and 0.855 and 0.873, respectively. CONCLUSIONS: V(5) and V(20) could act as predictors for RP; however, single DVH metrics did not appear to have high predictive power for RP.

The role of superoxide anion in the regulation of epidermal growth factor or the expression and proliferation of its receptor in prostate cancer cell line PC3.

The purpose of this study was to investigate the role of superoxide anion(O2-) in the regulation of epidermal growth factor (EGF) or epidermal growth factor receptor (EGFR) expression and proliferation in the prostate cancer cell line PC3. Cell proliferation was tested by a 3-(4,5-dimethylthiazol-2-yl)-diphenyltetrazolium bromide (MTT) assay in the presence of O2-, EGF or their combination. Immunohistochemistry was carried out to assay the expression of EGF or EGFR. EGF or EGFR mRNA expression in the cells treated with O2- was examined by in situ hybridisation. The proliferation was significantly inhibited by O2- in a concentration-dependent manner ranging from 9 to 36 micromol/l nicotinamide adenine dinucleotide (NADH) combined with 2-8 micromol/l N-methylphenazonium methyl sulfate (PMS). The enhancement of proliferation induced by 5 ng/ml EGF was significantly overcome by O2-. Although O2- was not able to alter EGFR mRNA expression, O2- at the concentration of 18 micromol/l NADH and 4 micromol/l PMS reduced EGFR protein expression. O2- at the concentration of 18 micromol/l NADH and 4 micromol/l PMS can downregulate EGF and EGF mRNA expression.