1.Study on DNA methylation in HEB cells exposed to PM(2.5) by application of methylation chip technology.
Run Bing LI ; Bing Yu WANG ; Shuang Jian QIN ; Xin Yun XU ; Zhao Hui ZHANG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2022;40(3):177-182
Objective: To screen the differential methylation sites, genes and pathways of air pollution fine particles (PM(2.5)) on human bronchial epithelial (HBE) cells by methylation chip and bioinformation technology, so as to provide scientific basis for further study of the toxicological mechanism of PM(2.5) on HBE cells. Methods: In August 2020, HBE cells were infected with 10 μg/ml and 50 μg/ml PM(2.5) aqueous solution for 24 h, namely PM(2.5) 10 μg/ml exposure group (low dose group) and PM(2.5) 50 μg/ml exposure group (high dose group) ; uninfected HBE cells were used as control group. The DNA fragments were hybridized with the chip, the chip scanned and read the data, analyzed the data, screened the differential methylation sites, carried out GO analysis and KEGG analysis of the differential methylation sites, and analyzed the interaction relationship of the overall differential methylation sites by functional epigenetic modules (FEMs). Results: Compared with the control group, 127 differential methylation sites were screened in the low-dose group, including 89 genes, including 55 sites with increased methylation level and 72 sites with decreased methylation level. The differential methylation sites were mainly concentrated in the Body region and UTR region. Compared with the control group, 238 differential methylation sites were screened in the high-dose group, including 168 genes, of which 127 sites had increased methylation level and 111 sites had decreased methylation level. The differential heterotopic sites were mainly concentrated in the Body region and UTR region. Through FEMs analysis, 8 genes with the most interaction were screened, of which 6 genes had significant changes in methylation level. MALT1 gene related to apoptosis was found in the heterotopic site of methylation difference in low-dose group; PIK3CA and ARID1A genes related to carcinogenesis were found in the heterotopic sites of methylation difference in high-dose group; TNF genes related to tumor inhibition were found in the results of FEMs analysis. Conclusion: After PM(2.5) exposure to HBE cells, the DNA methylation level is significantly changed, and genes related to apoptosis and carcinogenesis are screened out, suggesting that the carcinogenic mutagenic effect of PM(2.5) may be related to DNA methylation.
Air Pollutants/toxicity*
;
Basic Helix-Loop-Helix Transcription Factors/analysis*
;
Carcinogenesis
;
DNA Methylation
;
Humans
;
Particulate Matter/toxicity*
;
Technology
2.Energy power in mountains: difference in metabolism pattern results in different adaption traits in Tibetans.
Zhen-Zhong BAI ; Guo-En JIN ; Tana WU-REN ; Qin GA ; Ri-Li GE
Chinese Journal of Applied Physiology 2012;28(6):488-493
Energy metabolism plays an important role in life survival for species living in high altitude hypoxia condition. Air-breathing organisms require oxygen to create energy. Tibetans are the well-adapted highlanders in Qinghai-Tibetan Plateau. It was thought that different metabolic approaches could lead to different adaptation traits to high altitude hypoxia. Recently identified hypoxia inducible factors pathway regulators, endothelial PAS domain protein1 (EPAS1)/HIF-2a and PPARA, were involved in decreasing hemoglobin concentrations in Tibetans. Because EPAS1 and PPARA also modulated the energy metabolism during hypoxia, we hypothesized that positive selected EPAS1 and PPARA genes were also involved in unique energy metabolisms in Tibetans. In this brief review, we take a look into genetic determinations to energy metabolisms for hypoxia adaptations traits in Tibetans and mal-adaptive conditions such as high altitude diseases.
Acclimatization
;
genetics
;
Altitude
;
Basic Helix-Loop-Helix Transcription Factors
;
metabolism
;
Energy Metabolism
;
Hemoglobins
;
analysis
;
Humans
;
Hypoxia
;
metabolism
;
Oxygen
;
metabolism
;
Phenotype
;
Tibet
3.Perivascular epithelioid cell tumor of the lung: a clinicopathological analysis of eight cases.
J LI ; R P HUANG ; P PANG ; X GUO ; Y H WANG ; L C GUO ; S HUANG
Chinese Journal of Pathology 2023;52(11):1126-1131
Objective: To investigate the clinicopathological features of perivascular epithelioid cell tumor (PEComa) of the lung. Methods: Eight PEComa cases of the lung diagnosed at the First Affiliated Hospital of Soochow University, Suzhou, China from July 2008 to December 2021 were collected and subject to immunohistochemical staining, fluorescence in situ hybridization and next generation sequencing. The relevant literature was reviewed and the clinicopathological features were analyzed. Results: There were 5 males and 3 females, aged from 18 to 70 years (mean 39 years). There were 3 cases of the right upper lung, 3 cases of the left lower lung, 1 case of the left upper lung and 1 case of the right middle lung. Seven cases were solitary and 1 case was multifocal (4 lesions). Seven cases were benign while one was malignant. The tumors were all located in the peripheral part of the lung, with a maximum diameter of 0.2-4.0 cm. Grossly, they were oval and well circumscribed. Microscopically, the tumor cells were oval, short spindle-shaped, arranged in solid nests, acinar or hemangiopericytoma-like patterns, with clear or eosinophilic cytoplasm. The stroma was rich in blood vessels with hyalinization. Coagulated necrosis and high-grade nuclei were seen in the malignant case, and calcification was seen in 2 cases. Immunohistochemically, the tumor cells were positive for Melan A (8/8), HMB45 (7/8), CD34 (6/8), TFE3 (4/7), and SMA (3/8). All cases were negative for CKpan and S-100. TFE3 (Xp11.2) gene fusion was examined using the TFE3 break-apart fluorescence in situ hybridization in 5 cases, in which only the malignant case was positive. The next generation sequencing revealed the SFPQ-TFE3 [t(X;1)(p11.2;p34)] fusion. Follow-up of the patients ranged from 12 to 173 months while one patient was lost to the follow-up. The malignant case had tumor metastasis to the brain 4 years after the operation and then received radiotherapy. Other 6 cases had no recurrence and metastasis, and all the 7 patients survived. Conclusions: Most of the PEComas of the lung are benign. When there are malignant morphological features such as necrosis, high-grade nuclei or SFPQ-TFE3 gene fusion, close follow-up seems necessary.
Male
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Female
;
Humans
;
In Situ Hybridization, Fluorescence
;
Perivascular Epithelioid Cell Neoplasms/pathology*
;
Lung/pathology*
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Basic Helix-Loop-Helix Leucine Zipper Transcription Factors/genetics*
;
Necrosis
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Biomarkers, Tumor/analysis*
4.Relationship between Hes-1 and airway inflammation and remodeling in a rat model of asthma.
Ba-Cuo-Zhen QIANG ; Liang XIE ; Han-Min LIU ; Zhi-Yu FAN ; Li-Na CHEN
Chinese Journal of Contemporary Pediatrics 2014;16(6):648-653
OBJECTIVETo study the changes of Hes-1, the target gene of Notch signaling pathway, and its relationship with airway inflammation and remodeling in a rat model of asthma.
METHODSForty-eight rats were randomly divided into an asthma group and a control group. The rats in the asthma group were sensitized and challenged by ovalbumin (OVA), and normal saline was used in the control group. Two groups were further divided into 3 subgroups according to time points after challenging, i.e. 4 weeks, 8 weeks and 12 weeks (n=8 rats each). Pathological changes of lungs were observed by light microscopy and the thickness of bronchial smooth muscle layer (Wam) was measured. The levels of IL-4 and INF-γ in rat serum and bronchoalveolar lavage fluids (BALF) were measured using ELISA. Expression levels of Hes-1 protein and mRNA were determined by immunohistochemistry and quantitative real-time PCR respectively.
RESULTSTogether with the extension of challenging, the Wam of rats in the asthma group increased, a decrease of INF-γ level and an increase of IL-4 level in serum and BALF were also observed, and the differences were statistically significant compared with those in the corresponding control group (P<0.05). Hes-1 protein and mRNA levels also increased gradually after OVA challenging and were higher than those in the control group (P<0.05). The levels of Hes-1 protein and mRNA were positively correlated with Wam and IL-4 in serum and BALF, but were inversely correlated with INF-γ in serum and BALF (P<0.05).
CONCLUSIONSLevels of Hes-1 protein and mRNA increased, which were closely related with the levels of airway inflammatory factors and remodeling of airway smooth muscle. Hes-1 may play an important role in the pathogenesis of asthma.
Airway Remodeling ; Animals ; Asthma ; etiology ; Basic Helix-Loop-Helix Transcription Factors ; analysis ; genetics ; physiology ; Disease Models, Animal ; Homeodomain Proteins ; analysis ; genetics ; physiology ; Interferon-gamma ; analysis ; Interleukin-4 ; analysis ; Male ; Rats ; Rats, Sprague-Dawley ; Transcription Factor HES-1
5.Effects of SCL antisense ooligonucleotides on K562 and CEM cell lines.
Zheng-Jin ZHENG ; Jian-Da HU ; Shu-Hua HUANG ; Shao-Yuan WANG ; Lian-Huang LU
Journal of Experimental Hematology 2002;10(5):404-408
The stem cell leukemia (SCL) gene is a new oncogene related with leukemogenesis. To explore the effects of antisense oligonucleotides of SCL on leukemic cells, SCL antisense phosphorothioate oligodeoxynucleotides (AS-PS-ODN) were used to treat K562 and CEM leukemic cell lines to observe the effects on proliferation, differentiation, apoptosis and SCL mRNA expression in the cells. The results showed that incubation of K562 or CEM cells with AS-PS-ODN at different concentrations led to inhibition of cell proliferation, and the inhibitory effects varied with the incubation time. The positive rate of benzidine staining in K562 cells increased significantly after the inhibition with AS-PS-ODN, compared with S-PS-ODN treatment. The characteristics of apoptosis were observed in K562 cells treated with AS-PS-ODN, but not in CEM cells. Expression of SCL mRNA in K562 and CEM cells and SIL-SCL mRNA in CEM cells decreased after incubation of AS-PS-ODN. It is concluded that SCL AS-PS-ODN inhibits specifically the proliferation of K562 and CEM cells, also decreases the level of SCL and SIL-SCL mRNA expression. AS-PS-ODN enhances erythroid differentiation and induces premature apoptosis in K562 cells.
Apoptosis
;
drug effects
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Basic Helix-Loop-Helix Transcription Factors
;
Cell Division
;
drug effects
;
DNA-Binding Proteins
;
antagonists & inhibitors
;
physiology
;
Humans
;
K562 Cells
;
Oligonucleotides, Antisense
;
pharmacology
;
Proto-Oncogene Proteins
;
antagonists & inhibitors
;
physiology
;
RNA, Messenger
;
analysis
;
T-Cell Acute Lymphocytic Leukemia Protein 1
;
Transcription Factors
;
antagonists & inhibitors
;
physiology
6.Expression of a novel bHLH-Zip gene in human testis.
Jia-Hao SHA ; Zuo-Min ZHOU ; Jian-Min LI ; Ming LIN ; Hui ZHU ; Hu ZHU ; Ya-Dong ZHOU ; Li-Long WANG ; Yi-Quan WANG ; Kai-Ya ZHOU
Asian Journal of Andrology 2003;5(2):83-88
AIMTo identify specifically expressed genes in the adult and fetal testes.
METHODSA human testis cDNA microarray was established. Then the mRNA of adult and fetal testis was purified and probes were prepared by a reverse transcription reaction with the testis mRNA as template. The microarray was hybridized with probes of adult and fetal testes. The nucleic sequences of differentially expressed genes were determined and homologies were searched in the databases of the GenBank.
RESULTSWhen hybridized with adult or fetal testis probes, the positive clones were 96.8 % and 95.4 %, respectively. Among these genes, one was a new testis-specific gene, which was named TSP1. TSP1 was highly expressed in human adult testis. The cDNA of TSP1 was 1,484 bp in length. The cDNA sequence of this clone was deposited in the Genbank (AF333098). TSP1 was also determined as Interim Gen Symbol (Unigene, No. Hs.98266). Protein analysis showed that TSP1 contained two functional domains: an N-terminal basic helix-loop-helix (bHLH) and a C-terminal leucine zipper (Zip). Homologous analysis showed that the 430 amino acid sequences deduced from the 1293 bp open reading frame (ORF) had a homology with the human gene FLJ2509 (AK098575). TSP1 had also a sequence homology with Spz 1 protein of mouse. Expression profiles showed that TSP1 was specifically and strongly expressed in the testis.
CONCLUSIONTSP1 is a gene highly expressed in adult testis. It may play an important role in spermatogenesis in the humans.
Adult ; Amino Acid Sequence ; genetics ; Base Sequence ; genetics ; Basic Helix-Loop-Helix Leucine Zipper Transcription Factors ; Fetus ; metabolism ; Gene Expression ; Genes ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Oligonucleotide Array Sequence Analysis ; Sequence Homology, Amino Acid ; Testis ; embryology ; metabolism ; Transcription Factors ; chemistry ; genetics ; metabolism
7.Study on clinicopathologic features and immunophenotype of 114 cases of renal cell carcinoma.
Hong ZOU ; Li-juan PANG ; Wen-hao HU ; Feng LI ; Hong-an LI ; Jin-fang JIANG ; Wei-hua LIANG ; Zhen-zhu SUN ; Chun WANG ; Jian-yong LANG
Chinese Journal of Pathology 2008;37(11):726-731
OBJECTIVETo study the clinicopathologic features and immunophenotype of renal cell carcinomas, and to discuss their diagnostic value.
METHODSThe clinicopathologic features of 114 cases of renal cell carcinoma were reviewed and categorized on the basis of 2004 WHO classification. Immunohistochemical study for a panel of antibodies (including CK, CD10, vimentin, CD117, AMACR, CK7 and TFE3) was carried out. The follow-up data, if available, were also analyzed.
RESULTSThe cases were reclassified into 5 subtypes, including 77 cases (67.5%) of clear cell carcinoma (CCRCC), 11 cases (9.6%) of papillary renal cell carcinoma (PRCC), 14 cases (12.3%) of chromophobe renal cell carcinoma (chrRCC), 10 cases (8.8%) of renal carcinoma associated with Xp11.2 translocations/TFE3 gene fusions (Xp11.2RCC) and 2 cases (1.8%) of unclassified renal cell carcinoma (unRCC). Immunohistochemical study showed that the expression rates of CK, CD10 and vimentin in CCRCC were 93.5% (72/77), 93.5% (72/77) and 75.3% (58/77), respectively. On the other hand, all the 11 cases of PRCC studied were positive for AMACR. The expression rate of CD117 in chrRCC was 78.5% (11/14). In the 10 cases of Xp11.2 RCC studied, the expression rates of TFE3, AMACR, CD10 and CK were 100% (10/10), 100% (10/10), 90% (9/10) and 70% (7/10), respectively.
CONCLUSIONSThe various subtypes of renal cell carcinomas are heterogeneous in histologic appearance and demonstrate distinctive immunophenotype. The expressions of CD10, vimentin, CD117, AMACR, CK7 and TFE3 are helpful in the differential diagnosis.
Adenocarcinoma, Clear Cell ; pathology ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Basic Helix-Loop-Helix Leucine Zipper Transcription Factors ; genetics ; immunology ; metabolism ; Biomarkers, Tumor ; analysis ; genetics ; Carcinoma, Papillary ; immunology ; pathology ; Carcinoma, Renal Cell ; immunology ; metabolism ; pathology ; Female ; Gene Fusion ; Humans ; Immunophenotyping ; Kidney Neoplasms ; immunology ; metabolism ; pathology ; Male ; Middle Aged ; Neprilysin ; analysis ; Racemases and Epimerases ; analysis ; genetics ; Translocation, Genetic ; Vimentin ; analysis ; World Health Organization ; Young Adult
8.Risk Factors for Metachronous Gastric Neoplasms in Patients Who Underwent Endoscopic Resection of a Gastric Neoplasm.
Hyuk YOON ; Nayoung KIM ; Cheol Min SHIN ; Hye Seung LEE ; Bo Kyoung KIM ; Gyeong Hoon KANG ; Jung Mogg KIM ; Joo Sung KIM ; Dong Ho LEE ; Hyun Chae JUNG
Gut and Liver 2016;10(2):228-236
BACKGROUND/AIMS: To identify the risk factors for metachronous gastric neoplasms in patients who underwent an endoscopic resection of a gastric neoplasm. METHODS: We prospectively collected clinicopathologic data and measured the methylation levels of HAND1, THBD, APC, and MOS in the gastric mucosa by methylation-specific real-time polymerase chain reaction in patients who underwent endoscopic resection of gastric neoplasms. RESULTS: A total of 257 patients with gastric neoplasms (113 low-grade dysplasias, 25 high-grade dysplasias, and 119 early gastric cancers) were enrolled. Metachronous gastric neoplasm developed in 7.4% of patients during a mean follow-up of 52 months. The 5-year cumulative incidence of metachronous gastric neoplasm was 4.8%. Multivariate analysis showed that moderate/severe corpus intestinal metaplasia and family history of gastric cancer were independent risk factors for metachronous gastric neoplasm development; the hazard ratios were 4.12 (95% confidence interval [CI], 1.23 to 13.87; p=0.022) and 3.52 (95% CI, 1.09 to 11.40; p=0.036), respectively. The methylation level of MOS was significantly elevated in patients with metachronous gastric neoplasms compared age- and sex-matched patients without metachronous gastric neoplasms (p=0.020). CONCLUSIONS: In patients who underwent endoscopic resection of gastric neoplasms, moderate/severe corpus intestinal metaplasia and a family history of gastric cancer were independent risk factors for metachronous gastric neoplasm, and MOS was significantly hypermethylated in patients with metachronous gastric neoplasms.
Aged
;
Basic Helix-Loop-Helix Transcription Factors/genetics
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DNA Methylation
;
Female
;
Gastrectomy/methods
;
Genes, APC/physiology
;
Genes, mos/genetics
;
Humans
;
Incidence
;
Male
;
Middle Aged
;
Multivariate Analysis
;
Neoplasms, Second Primary/epidemiology/*genetics/pathology
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Proportional Hazards Models
;
Risk Factors
;
Stomach Neoplasms/genetics/*pathology/surgery
;
Thrombomodulin/genetics
9.Identification of a Novel Deletion Region in 3q29 Microdeletion Syndrome by Oligonucleotide Array Comparative Genomic Hybridization.
Eul Ju SEO ; Kyung Ran JUN ; Han Wook YOO ; Hanik K YOO ; Jin Ok LEE
The Korean Journal of Laboratory Medicine 2010;30(1):70-75
BACKGROUND: The 3q29 microdeletion syndrome is a genomic disorder characterized by mental retardation, developmental delay, microcephaly, and slight facial dysmorphism. In most cases, the microdeletion spans a 1.6-Mb region between low-copy repeats (LCRs). We identified a novel 4.0- Mb deletion using oligonucleotide array comparative genomic hybridization (array CGH) in monozygotic twin sisters. METHODS: G-banded chromosome analysis was performed in the twins and their parents. Highresolution oligonucleotide array CGH was performed using the human whole genome 244K CGH microarray (Agilent Technologies, USA) followed by validation using FISH, and the obtained results were analyzed using the genome database resources. RESULTS: G-banding revealed that the twins had de novo 46,XX,del(3)(q29) karyotype. Array CGH showed a 4.0-Mb interstitial deletion on 3q29, which contained 39 genes and no breakpoints flanked by LCRs. In addition to the typical characteristics of the 3q29 microdeletion syndrome, the twins had attention deficit-hyperactivity disorder, strabismus, congenital heart defect, and gray hair. Besides the p21-activated protein kinase (PAK2) and discs large homolog 1 (DLG1) genes, which are known to play a critical role in mental retardation, the hairy and enhancer of split 1 (HES1) and antigen p97 (melanoma associated; MFI2) genes might be possible candidate genes associated with strabismus, congenital heart defect, and gray hair. CONCLUSIONS: The novel 4.0-Mb 3q29 microdeletion found in the twins suggested the occurrence of genomic rearrangement mediated by mechanisms other than nonallelic homologous recombination. Molecular genetic and functional studies are required to elucidate the contribution of each gene to a specific phenotype.
Adaptor Proteins, Signal Transducing/genetics
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Adolescent
;
Attention Deficit Disorder with Hyperactivity/genetics
;
Basic Helix-Loop-Helix Transcription Factors/genetics
;
*Chromosome Deletion
;
Chromosome Disorders/*genetics
;
*Chromosomes, Human, Pair 3
;
Comparative Genomic Hybridization/*methods
;
Diseases in Twins/*genetics
;
Female
;
Homeodomain Proteins/genetics
;
Humans
;
In Situ Hybridization, Fluorescence
;
Melanoma-Specific Antigens/genetics
;
Membrane Proteins/genetics
;
Oligonucleotide Array Sequence Analysis
;
Syndrome
;
Twins
;
p21-Activated Kinases/genetics
10.Correlations of hypoxia-inducible factor-1α/hypoxia-inducible factor-2α expression with angiogenesis factors expression and prognosis in non-small cell lung cancer.
Xian-Hua WU ; Cheng QIAN ; Kai YUAN
Chinese Medical Journal 2011;124(1):11-18
BACKGROUNDHypoxia-inducible factor (HIF) may play an important role in the process of tumorigenesis as well as tumor progression. The aim of this study was to compare the expression between HIF-1α and HIF-2α in tumor angiogenesis and the overall impact on patient prognosis in human non-small cell lung cancer (NSCLC).
METHODSIn the current work we compared the immunohistochemical expression of HIF-1α and HIF-2α in surgical specimens of 140 patients with NSCLC in a tissue microarray study. Relationships between HIF-α expression and clinicopathological or angiogenic factors, including prognosis, were analyzed.
RESULTSHigh HIF-1α and HIF-2α expression was noted in 49/140 (35.0%) and in 64/140 (45.7%) of the cases, respectively. There was no direct correlation between HIF-1α and HIF-2α expression. Patients with advanced stage tumors had frequent high expression of HIF-2α (P = 0.007), and we also found a significant correlation between HIF-2α and T or N stage (P = 0.030 and 0.043, respectively). HIF-1α showed a marginal association with T stage (P = 0.084), which showed a higher expression in early stage tumors. A significant correlation (P = 0.045) was noticed between HIF-1α and vascular endothelial growth factor (VEGF) expression while the expression levels of thymidine phosphorylase (TP), cyclooxygenase (COX)-2 and microvessel density (MVD) were significantly higher in high HIF-2α tumors (P = 0.020, 0.004, and 0.046, respectively). In addition, univariate analysis of overall survival demonstrated that HIF-2α expression, but not HIF-1α, was related to poor outcome (P = 0.001) and it retained significant in multivariate analysis (P = 0.036).
CONCLUSIONSTaken together, we conclude that HIF-1α and HIF-2α may differentially regulate the major angiogenic factors in different stages of the tumor process in NSCLC. HIF-2α may play a dominant role in tumor angiogenesis and appears to be of obvious value as a significant prognostic factor in NSCLC.
Adult ; Aged ; Angiogenesis Inducing Agents ; metabolism ; Basic Helix-Loop-Helix Transcription Factors ; metabolism ; Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Cyclooxygenase 2 ; metabolism ; Female ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Immunohistochemistry ; Male ; Microvessels ; Middle Aged ; Multivariate Analysis ; Prognosis ; Thymidine Phosphorylase ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism