Suppression of the SLC7A11/glutathione axis causes ferroptosis and apoptosis and alters the mitogen-activated protein kinase pathway in nasopharyngeal carcinoma.

SLC7A11 is a unit of the glutamate cystine antiporter Xc- system. It functions to import cystine for glutathione biosynthesis and maintains the redox balance in cells. Sorafenib inhibits the transporter activity of SLC7A11. The use of sorafenib has been approved in the treatment of multiple cancers. However, at present, our understanding of the mechanism of SLC7A11 and sorafenib in nasopharyngeal carcinoma (NPC) remains limited. We found that the expression of SLC7A11 was upregulated in NPC. A high SLC7A11 expression was associated with poor prognosis, metastasis, and an advanced T stage, which can be used as an independent prognostic indicator of NPC. In vitro, we observed that NPC cells relied on cystine for survival. Targeting SLC7A11 resulted in glutathione biosynthesis limitation, intracellular reactive oxygen species accumulation, lipid peroxides, ferroptosis, and apoptosis. Meanwhile, it altered mitogen activated protein kinase pathway, including p38 activation but ERK inhibition in NPC. This limited the proliferation of NPC cells. Sorafenib inhibited the proliferation and induced the death of NPC cells in vivo. In conclusion, SLC7A11 plays an important role in the occurrence and progression of NPC and may be a novel target for NPC treatment.

Convergent genomic signatures of high-altitude adaptation among domestic mammals.

Abundant and diverse domestic mammals living on the Tibetan Plateau provide useful materials for investigating adaptive evolution and genetic convergence. Here, we used 327 genomes from horses, sheep, goats, cattle, pigs and dogs living at both high and low altitudes, including 73 genomes generated for this study, to disentangle the genetic mechanisms underlying local adaptation of domestic mammals. Although molecular convergence is comparatively rare at the DNA sequence level, we found convergent signature of positive selection at the gene level, particularly the EPAS1 gene in these Tibetan domestic mammals. We also reported a potential function in response to hypoxia for the gene C10orf67, which underwent positive selection in three of the domestic mammals. Our data provide an insight into adaptive evolution of high-altitude domestic mammals, and should facilitate the search for additional novel genes involved in the hypoxia response pathway.

GRK5 functions as an oncogenic factor in non-small-cell lung cancer.

Lung cancer is the leading cause of cancer-related deaths worldwide, and non-small-cell lung cancer (NSCLC) accounts for about 80% of all cases, which is the major subgroup of lung cancer. G protein-coupled receptor kinase 5 (GRK5) has been demonstrated to play pivotal roles in both development and progression of several pathological conditions including cancer. Here, we found that GRK5 expression was significantly increased in 539 NSCLC cancerous tissues than that in 99 normal non-cancerous tissues by immunohistochemistry analysis; we also showed intensive higher positive staining percentage in female and adenocarcinoma (ADC) NSCLC patients than that in male and squamous cell carcinoma (SCC) patients, respectively. In addition, GRK5 high expression NSCLC patients had a worse overall survival rate than the low expression patients. We provided evidence showing that both the mRNA and protein expression levels of GRK5 were increased in NSCLC cancerous cell lines (GLC-82, SPC-A-1, H520, H838, H358, A549, and H1299) comparing with that in normal human bronchial epithelium cell line (BEAS-2B), and identified many GRK5 mutations in NSCLC cancerous tissues. In addition, we found that depletion of GRK5 inhibited NSCLC cancerous cell proliferation, migration in vitro, and xenograft tumor formation in vivo. Furthermore, GRK5 knockdown promoted cell cycle arrest at G2/M phase and induced cellular apoptosis. In summary, our data reveal an oncogenic role of GRK5 in NSCLC progression, indicating that GRK5 could be used as a new therapeutic target in future.

A Normalization-Free and Nonparametric Method Sharpens Large-Scale Transcriptome Analysis and Reveals Common Gene Alteration Patterns in Cancers.

Heterogeneity in transcriptional data hampers the identification of differentially expressed genes (DEGs) and understanding of cancer, essentially because current methods rely on cross-sample normalization and/or distribution assumption-both sensitive to heterogeneous values. Here, we developed a new method, Cross-Value Association Analysis (CVAA), which overcomes the limitation and is more robust to heterogeneous data than the other methods. Applying CVAA to a more complex pan-cancer dataset containing 5,540 transcriptomes discovered numerous new DEGs and many previously rarely explored pathways/processes; some of them were validated, both in vitro and in vivo, to be crucial in tumorigenesis, e.g., alcohol metabolism (ADH1B), chromosome remodeling (NCAPH) and complement system (Adipsin). Together, we present a sharper tool to navigate large-scale expression data and gain new mechanistic insights into tumorigenesis.

Rare coexistence of mediastinal hepatoid adenocarcinoma, idiopathic azoospermia and horseshoe kidney: a case report and review of the literature.

Hepatoid adenocarcinoma (HAC) is the term proposed for a special type of extrahepatic tumors, which is similar to the hepatocellular carcinoma (HCC) both in the histopathology and immunohistochemistry. HAC has been observed in the stomach, colon, pancreas, gall bladder, lung and female genital tract, but rarely in the mediastinum. Now we describe a case of a 28-year-old Chinese male with primary mediastinal HAC with lung and liver metastasis. In this patient, HAC was associated with horseshoe kidney and idiopathic nonobstructive azoospermia. It seemed derivation abnormalities during organogenesis in the embryo stage played a significant role in the pathogenesis of HAC, horseshoe kidney and idiopathic nonobstructive azoospermia. Even the pathogenesis was still unknown; it may merit consideration of HAC together with horseshoe kidney and idiopathic nonobstructive azoospermia as a syndrome rather than as a spectrum of coincidental diseases. Furthermore, we found the HAC is a neoplasm with unfavorable outcomes despite aggressive and multi-protocol strategies. The serum alpha fetoprotein (AFP) should be regarded as a useful marker for diagnostic purposes and therapeutic response evaluation of HAC.

Interruption of nuclear localization of ATBF1 during the histopathologic progression of head and neck squamous cell carcinoma.

BACKGROUND: The AT-motif binding factor 1 (ATBF1) gene is frequently altered at the genetic level in several types of cancer, but its protein expression and subcellular localization have not been well studied in human cancers, including head and neck squamous cell carcinomas (HNSCCs). METHODS: ATBF1 expression and localization were examined in 5 cell lines and 197 clinical specimens of HNSCC, and correlated with pathologic and clinical characteristics. RESULTS: ATBF1 was predominantly localized in the nucleus of hyperplastic squamous epithelium. Whereas nuclear ATBF1 dramatically decreased in invasive tumors (p = .0012), cytoplasmic ATBF1 levels progressively increased from dysplasia to invasive tumors (p < .0001), and the increase correlated with poor survival. Reduced nuclear ATBF1 level was also detected in HNSCC cell lines. CONCLUSIONS: Nuclear localization of ATBF1 is frequently interrupted in HNSCC, and the interruption is significantly associated with the progression of HNSCC. The cytoplasmic ATBF1 level could be useful for predicting patient survival.

Thyroid teratoma in an 11-month-old infant.

We report a case of congenital benign thyroid teratoma in an 11-month-old male infant who was found to have right thyroid gland mass since birth. The tumor was 25 x 20 x 15 mm with whole thin capsule and could be easily dissected from the surrounding normal thyroid tissue at surgery. Histologically, tumor had mature derivatives of the three primordial germ layers with a variety of benign and well-differentiated elements. It was the most conspicuous feature that the tumor was composed mainly of the neurological tissue resembling brain tissue with glial cells and ependymal epithelium components. There were a few anastomosing variably sized tubules and cysts lined by ependymal epithelial cells with papillary feature and retinal pigment epithelial cells. In summary, benign teratoma of thyroid gland in an 11-month-old infant was morphologically and immunophenotypically identified.

[Effect of all-trans-retinoic acid on C6 glioma cell proliferation and differentiation].

OBJECTIVE: To explore the effect of all-trans-retinoic acid (ATRA) on the growth inhibition and cellular differentiation of C6 glioma cells. METHODS: Human glioma C6 cells were treated with 5 mg/L ATRA,and the inhibition of cell growth was assessed by methyl thiazolyl tetrazolium assay. The differentiation of C6 cells was determined by flow cytometry, microscopy,transmission electron microscope, and immunohistochemical technique. RESULTS: Treatment of ATRA could result in the growth inhibition of C6 cells, and the cell density significantly decreased(P<0.01). The cell cycle distribution was changed, G0/G1 phase was prolonged, and cells at S phase decreased(P<0.01). The C6 glioma cells displayed normal fibroblast-like morphology under the microscope before the induction, and the ATRA-treated C6 cells became slightly long, turned into round in the middle, and had protrusions at both ends. The ATRA-treated C6 cells did not display obvious apoptosis by flow cytometry(P>0.05).Whereas, early apoptosis was observed under the transmission electron microscope, the vacuoles increased, the mitochondria and endoplasmic reticulum were abundant in the cytoplasm, and the cellular structures tended to be normal.The expression of glial fibrillaryacidic protein in C6 cells increased in the treatment group. CONCLUSION: ATRA can inhibit the proliferation, and induce the differentiation of C6 glioma cells.

Glial choristoma of the tongue: report of a case and review of the literature.

Glial choristoma of the tongue is an extremely rare developmental malformation. The authors report a case of a 5-month-old male baby with a congenital glial choristoma located on the posterior part of midline of the left dorsal tongue. Histological examination of the resected specimen revealed a poorly demarcated submucosal mass containing neuroglial tissue, scattered neuron, choroids plexus and ependyma. In addition to neuroglial tissue, a sheet of leptomeningeal tissue was observed more rarely in the case. The clinical and pathological characteristics of previous cases and their probable embryogenesis were also reviewed.

Gene expression profiling of nasopharyngeal carcinoma reveals the abnormally regulated Wnt signaling pathway.

Nasopharyngeal carcinoma (NPC) is a particularly common malignant disease in areas of south China and Southeast Asia. To characterize the gene expression profiling of NPC, we detected the gene expression profiles in 22 NPC and 10 nontumor nasopharyngeal epithelial tissues by complementary DNA microarray. We identified 503 genes that were significantly (P < .001) differentially regulated between NPC and nontumor nasopharyngeal epithelial tissues. The differentially expressed genes are involved in many signaling pathways, such as the Wnt, transforming growth factor-beta, and mitogen-activated protein kinase signaling pathways. The aberrant expression of the Wnt signaling pathway components, such as wingless-type MMTV integration site family, member 5A, Frizzled homolog 7, casein kinase IIbeta, beta-catenin, CREB-binding protein, and Dishevelled-associated activator of morphogenesis 2 was validated on the NPC tissue microarrays. The data suggest that the Wnt signaling pathway may be abnormally regulated in NPC, which provides insight into the molecular mechanisms of NPC.

[Expression of VEGF-C and angiogenesis, and lymphangiogenesis in papillary thyroid carcinoma].

OBJECTIVE: To investigate the relationship between the expression of vascular endothelial growth factor C (VEGF-C) and angiogenesis and lymphangiogenesis in papillary thyroid carcinoma (PTC). METHODS: Seventy-two PTC cases were divided into 3 groups according to the level of invasion: papillary microcarcinoma group (PMC group), intrathyroid carcinoma group (IPC group), and extrathyroid carcinoma group (EPC group). They were again divided into 2 groups according to lymph node metastasis: lymph node metastasis group and lymph node no-metastasis group. The expressions of VEGF-C, CD105 and vascular endothelial growth factor receptor-3 (VEGFR-3) were detected by SP method of immunohistochemical staining. The expression of VEGF-C was analyzed quantitatively by image analysis system, and the PI of VEGF-C (VEGF-C-PI), the number of MVD (microvessel density), and LVD (lymphaticvessel density) were obtained. RESULTS: The VEGF-C-PI of lymph node metastasis group (23.15 +/- 3.75) was higher than that of lymph node non-metastasis group (14.54 +/- 2.93) (P <0.01). MVD was 35.25 +/- 2.06 in the PMC group, 41.75 +/- 5.46 in the IPC group, and 52.58 +/- 4.16 in the EPC group, which showed the elevatory tendency with the increase of invasion (P < 00.5). LVD was 6.00 +/- 0.81 in the PMC group, 13.80 +/- 1.81 in the IPC group, and 19.17 +/- 2.96 in the EPC group, which again showed the elevatory tendency with the increase of invasion (P <0.05). The LVD of lymph node metastasis group (19.56 +/- 2.45) was significantly higher than that of lymph node non-metastasis group (12.48 +/- 2.84) (P < 0.05). VEGF-C was positively correlated with MVD and LVD (r = 0.743, 0.90, P <0.01). CONCLUSION: The expressions of VEGF-C and LVD are related to lymph node metastasis of PTC. MVD and LVD are related to the invasion of PTC. VEGF-C may play an important role in the angiogenesis and lymphangiogenesis.

[Anatomic study of the hypoglossal nerve in hypoglossal-facial nerve anastomosis].

OBJECTIVE: To determine the optimal position of hypoglossal nerve in hypoglossal-facial nerve anastomosis and the eligibility of hypoglossal-facial nerve anastomosis with the cervical loop. METHODS: The cervical course and adjacent structures of the hypoglossal nerve were observed on 21 adult cadavers. The hypoglossal nerve and facial nerve were taken from 3 fresh specimens, and the number of the fasciculus and the cross-sectional area of the nerve were measured. RESULTS: The facial nerve trunk were monofascicular with a cross-sectional area of 5.1-/+0.2 (range 4.6-5.7) mm(2). The number of the fasciculus and the cross-sectional areas of the nerve trunk and the fasciculus were 1.6-/+0.8 (range 1-4) mm(2) , 7.5-/+0.7 mm(2) (range 6.8-8.0) mm(2), and 4.7-/+0.6 (4.1-5.5) mm(2), respectively, at the proximal segment of the hypoglossal nerve, 3.6-/+0.5 (1-5) mm(2) , 5.6-/+0.5 (4.9-6.1) mm(2) , and 1.6-/+0.4 (0.9-2.2) mm(2) at the distal segment, and 2.4-/+0.8 (1-3) mm(2), 1.1-/+0.7 (0.6-2.2) mm(2), and 0.5-/+0.3 (0.3-1.2) mm(2) at the cervical loop. CONCLUSION: The cervical loop is inadequate for facial nerve anastomosis and the proximal segment is large enough to allow partial harvesting of the hypoglossal nerve for neurotisation of the facial nerve.

BRD2 is one of BRD7-interacting proteins and its over-expression could initiate apoptosis.

BRD7 is a potential nuclear transcription regulation factor related to nasopharyngeal carcinoma (NPC). BRD2, a putative BRD7-interacting protein, has been screened from human fetal brain cDNA library by yeast two-hybrid system. This study was to further identify the interaction between BRD7 and BRD2 in mammalian cells, and to investigate the subcellular localization of BRD2, as well as the effect on the functions of cell biology. Both immunoprecipitation and subcellular colocalization were performed together to identify the interaction of BRD7 with full-length BRD2, as well as C-terminal truncated BRD2 or N-terminal truncated BRD2. GFP direct fluorescence and Hochest 33258 staining were used to investigate the cellular localization pattern of BRD2 and the roles in initiating cell apoptosis in COS7 and HNE1. The results showed that BRD7 could interact with BRD2 and the region from amino acid 430 to 798 of BRD2 was critical for the interaction of BRD2 with BRD7. BRD2 mainly localizes in nucleus in two distribution patterns, diffused and dotted, and BRD2 has distinct roles in initiating apoptosis, and the dotted distribution pattern of BRD2 in nucleus may be a morphologic marker of cell apoptosis.

Differential expression of Epstein-Barr virus-encoded RNA and several tumor-related genes in various types of nasopharyngeal epithelial lesions and nasopharyngeal carcinoma using tissue microarray analysis.

Studies have revealed that Epstein-Barr virus (EBV) infection, genetic aberration, and environmental factors are of importance in the development of nasopharyngeal carcinoma (NPC), although the definite mechanism remains to be fully elucidated. The aim of our study is to investigate using tissue microarray analysis whether differential expression of EBV-encoded small RNA-1 (EBER-1) and several tumor-related genes were associated with NPC carcinogenesis. Immunohistochemistry and in situ hybridization were performed on tissue microarrays containing 148 NPCs and 164 noncancerous nasopharyngeal epithelia (NPE) with different morphologic features. We found that overexpressions of EBER-1 hybridization signals, p53, p21ras, and bcl-2 proteins and loss expressions of p16 and p27 proteins were significantly increased in NPC tissues compared with normal NPE and hyperplastic NPE (P

Effect of Yangqixue Qufengshi Recipe on rheumatoid arthritis model mice under different genetic backgrounds.

OBJECTIVE: To study the effect of Yangqixue Qufengshi Recipe (YQXQFS) on rheumatoid arthritis (RA) model mice under different genetic backgrounds. METHODS: Collagen Induced Arthritis (CIA) were established on HLA-DR4 transgenic (TG) mice and non-transgenic (NTG) mice, which partly were raised with YQXQFS, and the onset day of CIA, the level of type II collagen (CII)-reactive antibodies and the pathological scores of CIA were assessed. RESULTS: Under HLA-DR4 TG background (compared with NTG mice), the earlier onset day of CIA (11.22 +/- 3.35 days vs 16.56 +/- 4.75 days, P < 0.05) and higher level of CII-reactive antibodies (0.2274 +/- 0.1390 microg/ml vs 0.1101 +/- 0.0560 microg/ml, P < 0.05) were observed, but the pathological scores of CIA remained unchanged. YQXQFS could not influence the onset day of CIA and the level of CII-reactive antibodies, but had a certain effect on the total pathological scores (6.56 +/- 3.43 scores vs 11.11 +/- 5.64 scores) and bone erosion (0.22 +/- 0.44 scores vs 1.67 +/- 1.50 scores) of CIA on NTG mice (P < 0.05), NTG YQXQFS group compared with NTG experimental group. CONCLUSION: YQXQFS had a certain effect on RA model, but had no significant effect on HLA-DR4 related CIA.

[Function of a novel brain-specific gene LRRC4].

OBJECTIVE: To study the suppressive effect of LRRC4 gene on human glioma U251 cells and further investigate its biological functions. METHODS: H&E, DNA and AgNORs stainings were performed on LRRC4-transfected U251 cells, mock-transfected U251 cells and non-transfected U251 cells, respectively. Quantitative analysis including cell morphometry, DNA content, DNA ploidy, silver stained argyrophilic nucleolar organizer regions (AgNORs) were investigated by image analysis. Flow cytometry was employed to determine the difference of cell cycle distribution and MTT staining was used to elucidate the activity of the LRRC4-transfected U251 cells. RESULTS: The morphological cell parameters such as area, perimeter and diameter, DNA content, chromosomal aneupoloidy, mean area of AgNORs particles and mean nucleus area of the LRRC4-transfected U251 cells were remarkably decreased compared to those of the mock-transfected and non-transfected U251 cells (P < 0.05, P < 0.01). Meanwhile, significant accumulation of cells in G(0)/G(1) phase but decrease of cells in S and G(2)/M phase, was observed in transfected U251 cells compared to those of the mock-transfected and non-transfected U251 cells (P < 0.05, P < 0.01). MTT staining showed that proliferation activity of both the mock- and non-trasfected U251 cells was significantly higher than that of the U251 cells transfected with LRRC4 gene (P < 0.01). CONCLUSION: LRRC4 gene might be involved in tumor suppression by restraining DNA synthesis and the nucleoli organizer regions-associated proteins, keeping the cell cycles in phase G(0)/G(1) and reducing proliferation activity of the glioma cells. Morphometry combined with other techniques such as flow cytometry and MTT staining can well elucidate the biological function of novel genes.

A susceptibility locus at chromosome 3p21 linked to familial nasopharyngeal carcinoma.

Nasopharyngeal carcinoma (NPC) poses one of the serious health problems in southern Chinese, with an incidence rate ranging from 15 to 50/100,000. Chromosome translocation t(1;3) and frequent loss of heterogeneity on short arms of chromosome 3 and 9 have been reported to be associated with NPC, and a genome-wide scan identified an NPC susceptibility locus on chromosome 4p15.1-q12 recently. In our study, we collected samples from 18 families at high risk of NPC from the Hunan province in southern China, genotyped with a panel of polymorphic markers on short arms of chromosomes 3, 9, and 4p15.1-q12. A locus on 3p21 was identified to link to NPC with a maximum logarithm of odds for linkage score of 4.18. Fine mapping located the locus to a 13.6-cM region on 3p21.31-21.2, where a tumor suppressor gene cluster resided. Our findings identified a novel locus for NPC and provided a map location for susceptibility genes candidates. In contrast to a recent study, no significant evidence for NPC linkage to chromosomes 4 and 9 was observed.

[Expression of matrix metalloproteinases and tissue inhibitors of matrix metalloproteinases in the hepatocellular carcinomas].

OBJECTIVE: To explore the expression and clinical prognostic significance of matrix metalloproteinase-2 mRNA (MMP-2 mRNA), tissue inhibitor of matrix metalloproteinase-2 mRNA (TIMP-2 mRNA), matrix metalloproteinase-2 protein (MMP-2), matrix metalloproteinase-9 protein (MMP-9), tissue inhibitor of matrix metalloproteinase-1 protein (TIMP-1), and tissue inhibitor of matrix metalloproteinase-2 protein (TIMP-2) in the hepatocellular carcinomas (HCCs). METHODS: Fifty-six specimens of HCCs from 56 patients, who were followed-up, were investigated by in situ hybridization with specific probes for MMP-2, TIMP-2, and by immunohistochemistry with anti-MMP-2, MMP-9 and anti-TIMP-1, TIMP-2 monoclonal antibody. We analyzed the data with chi-square test, spearmans correlation analysis, monovariate Kaplan-Meier plot and multivariate Cox regression analysis. RESULTS: 1. The positive expression of MMP-2mRNA, TIMP-2mRNA, MMP-2 protein, MMP-9 protein, TIMP-1 protein and TIMP-2 protein in the 56 HCCs cases were 48 (85.7%), 35 (62.5%), 44 (78.6%), 41 (73.2%), 30 (53.6%), and 38 (68%), respectively. 2. We found over-expression of MMP-2 mRNA, MMP-2 protein, and MMP-9 protein, but low expression of TIMP-1 protein in the 56 cases of HCCs (P < 0.01, P < 0.05). 3. There was a positive association between TIMP-2mRNA and TIMP-2 protein expression, and between MMP-2 mRNA and MMP-2 protein in HCCs, respectively (r = 0.316, P < 0.05; r = 0.356, P < 0.05). 4. Over-expression of MMP-2 mRNA was positively correlated to the tumor size and TNM classification (r = 0.441, P < 0.001; r = 0.340, P < 0.05), and MMP-9 protein was related to shortened survival (P < 0.05). 5. In both monovariate Kaplan-Meir plot and multivariate Cox regression analysis, the expression of MMP-2 protein and MMP-9 protein were linked to unfavorable prognosis. These results were further confirmed by multivariate analysis in which MMP-2 protein and MMP-9 protein emerged as independent prognostic factors for poor survival regardless of the age, tumor size, tumor grades, TNM classification and expression of MMP-2mRNA, TIMP-2mRNA, TIMP-1 protein and TIMP-2 protein. The hazard ratios of expression of MMP-2 protein and MMP-9 protein were 3.875 and 4.528, respectively. CONCLUSION: The over-expression of MMP-2mRNA, MMP-2 protein and MMP-9 protein and the imbalance between MMP-2 and TIMP-2 play pivotal roles in the degradation of excellular matrix of HCCs. MMP-2 and MMP-9 immunoreactive protein have been closely related to a shortened survival independent of major prognostic indicators in the primary HCC and increase the risk of the patients after the operation.

[Genetic polymorphism of 8 STR loci on short arm of chromosome 3].

OBJECTIVE: To get the genotype and allele frequency distributions of 8 short tandem repeat (STR) loci on chromosome 3p (D3S1297, D3S1489, D3S1266, D3S1568, D3S1289, D3S1300, D3S1285 and D3S3681) in Chinese Han population in Hunan area. METHODS: Blood samples were collected from the random Han individuals in Hunan and the whole genomic DNA was extracted. STR loci were amplified by multiplex-PCR technique and genotyped by ABI 377 sequencer. RESULTS: Ninety-one alleles were detected, with frequencies ranging from 0.002 to 0.431, and these alleles constituted 312 genotypes. All the 8 loci met Hardy-Weinberg equilibrium. The statistical analysis of 8 STR loci showed the heterozygosity (H) >or= 0.729, the discrimination power (DP) >or= 0.725, the probabilities of paternity exclusion (PPE) >or= 0.596, and the polymorphic information content (PIC >or= 0.682). The result indicated that there was a significant difference between Han ethnic group and the white and the black. CONCLUSION: These results could serve as valuable data to enrich the Chinese genetic database and play an important role in Chinese population genetic and forensic medical application.