Interferon-related gene array in predicting the efficacy of interferon therapy in chronic hepatitis B / 生物医学工程学杂志

This study aims to clarify host factors of IFN treatment in the treatment of chronic hepatitis B (CHB) patients by screening the differentially expressed genes of IFN pathway CHB patients with different response to interferon (IFN) therapy. Three cases were randomly selected in IFN-responding CHB patients (Rs), non-responding CHB patients (NRs) and healthy participants, respectively. The human type I IFN response RT 2 profiler PCR array was used to detect the expression levels of IFN-related genes in peripheral blood monocytes (PBMCs) from healthy participants and CHB patients before and after Peg-IFN-α 2a treatment. The results showed that more differentially expressed genes appeared in Rs group than NRs group after IFN treatment. Comparing with healthy participants, IFNG, IL7R, IRF1, and IRF8 were downregulated in both Rs and NRs group before IFN treatment; CXCL10, IFIT1, and IFITM1 were upregulated in the Rs; IL13RA1 and IFI35 were upregulated in the NRs, while IFRD2, IL11RA, IL4R, IRF3, IRF4, PYHIN1, and ADAR were downregulated. The expression of IL15, IFI35 and IFI44 was downregulated by 4.09 ( t = 10.58, P < 0.001), 5.59 ( t = 3.37, P = 0.028) and 10.83 ( t = 2.8, P = 0.049) fold in the Rs group compared with the NRs group, respectively. In conclusion, IFN-response-related gene array is able to evaluate IFN treatment response by detecting IFN-related genes levels in PBMC. High expression of CXCL10, IFIT1 and IFITM1 before treatment may suggest satisfied IFN efficacy, while high expression of IL13RA1, IL15, IFI35 and IFI44 molecules and low expression of IFRD2, IL11RA, IL4R, IRF3, IRF4, PYHIN1 and ADAR molecules may be associated with poor IFN efficacy.

Mutation analysis of the ABCA12 gene in two families with autosomal recessive congenital ichthyosis / 中华皮肤科杂志

Objective To detect mutations of the ABCA12 gene in 2 Chinese families with autosomal recessive congenital ichthyosis (ARCI).Methods According to the typical clinical manifestations,two probands were diagnosed with ARCI.DNA was extracted from the peripheral blood samples collected from the patients and their parents.High-throughput sequencing was conducted by using multi-gene array for genetic skin disorders to determine mutation sites in the probands,and then DNA isolated from the probands and their parents were bidirectionally verified by Sanger sequencing.Results Two compound heterozygous mutations (c.2759A＞G and c.7004A＞G) in the ABCA12 gene were found in the proband 1,and another two compound heterozygous mutations (c.6163_6164insT and c.7406G＞A) were identified in the proband 2.The parents of the two probands were heterozygous carriers of one of the two mutations in the ABCA12 gene.Function prediction for the 4 mutations showed that all of the 3 missense mutations (c.2759A＞G,c.7004A＞G and c.7406G＞A) may exert pathogenic effect,and fragnin encoded by the frameshift mutation c.6163_6164insT may also affect protein function,c.2759A＞G and c.6163_6164insT were newly identified mutation sites.Conclusion The compound heterozygous mutations in the ABCA 12 gene are the causative mutations responsible for ARCI in the two probands of the two pedigrees.

Effects of epigallocatechin-3-gallate on the differential expression of apoptosis-associated genes in human gastric cancer MGC-803 cells / 中国肿瘤临床

Objective: This study investigates the biological effects and explores the molecular mechanisms of epigallocate-chin-3-gallate (EGCG) on the apoptosis of the human gastric cancer MGC-803 cells. Methods: After treatment with EGCG, cell apopto-sis was verified by flow cytometry with Annexin V and propidium iodide staining, DNA agarose gel electrophoresis, and transmission electron microscopy. The expression profiles of the apoptosis-related genes in the MGC-803 cells with or without treatment by EGCG for 12 h (100 μmol/L), was identified using SuperArray Human Apoptosis Gene Array. The upregulated Fas-L gene and down-regulated Bag-1 gene were confirmed by reverse transcription polymerase chain reaction (RT-PCR) and Western blot. Results: When the MGC-803 cells were treated with EGCG at 25, 50, 100, and 200 μmol/L for 24 h, evident sub-diploid peaks were observed. Under treat-ment with 100 μmol/L for 4, 8, 12, and 24 h, the number of early apoptotic cells was greatly increased. When the cells were treated with 100 μmol/L for 24 h, the DNA extracted from the cells displayed a characteristic ladder pattern with agarose gel electrophoresis. Typi-cal morphological changes were observed by electron microscopy, including cell shrinkage, karyo-pyknosis, and the formation of apop-totic bodies. The differential expressions of eight apoptosis-associated genes were determined by gene array detection. The results of Fas-L and Bag-1 selected for RT-PCR and Western blot were consistent with those of gene array. Conclusion: EGCG induces apoptosis in MGC-803 cells, which might be mediated by a number of specific genes and various signal transduction pathways.

Mouse testicular gene expression pattern differences between spermatogonial stem cell proliferative and differential stages / 中华泌尿外科杂志

Objective To detect the mouse testicular gene expression pattern differences be-tween spermatogonial stem cell (SSC) proliferative and differential stages and study the molecular reg-ulation mechanism in SSC proliferation and differentiation. Methods With the interval of 24 days, male Kunming mice were injected intraperitoneally with two doses of busulfan (10 mg/kg) to establish spermatogenesis regeneration models. 36 k Mouse Genome Array was used to detect the differential gene expression profiles between the stages of SSC proliferation and differentiation. Bioinforrnsties analysis was conducted in GO (gene ontology) and KEGG (Kyoto Encyclopedia of Genes and Ge-nomes) pathway to describe the potential roles that may play in spermatogonial stern cells behavior regulation. Results Nine hundred and eleven differential expression genes were identified by gene arrays in mice testes, consisting of 608 up-regulated and 303 down-regulated in SSC proliferation stage and SSC differentiation stage. The differential expression genes were classified by their biological process, molecular function and cellular component, respectively. Alterations with statistical signifi-cance (P<0.05)appeared in 84 KEG(;signal pathways, including Notch and Wnt signaling pathways which had been proved to be important for stem cell maintenance. Fifty-six differential expression genes were selected as genes related to stem cells, among which 40 genes were up-regulated, including some stem cell biomarkers(such as Cd9, StraS, hgbl-, Oct4 and Thyl)and some growth factors(such as Fgf2, Pdgfa and Csfl). Conclustion The regulation of SSC proliferation and differentiation involves inmany differentially expressed genes in various signal pathways. This study provides a molecular basis for the elucidation of the molecular mechanism behind self-renewal and differentiation of spermatogonial stem cells.

Cell Cycle and Gene Expression of Adriamycin-Induced Apoptosis in Human Cervical Carcinoma HeLa cells / 대한산부인과학회잡지

OBJECTIVE: This work was demonstrated the induction of apoptosis in response to adriamycin, we checked the cell cycle of adriamycin-induced apoptosis and to investigate whether differential expression is associated with adriamycin-induced genes in human cervical carcinoma HeLa cells. METHODS: Apoptosis was measured by flow cytometry for cell cycle analysis in Hela cells. Differential expression is associated with adriamycin-induced genes in HeLa cells, it was performed to purifiy the RNA, cDNA probe and hybridization. The various different overexpressed genes were determined by gene array analysis (GDA). Analysis were referenced Incyte Genomics Co. (http://www.ncbi.nlm.nih.gov/). RESULTS: We found that adriamycin was induced apoptosis in a dose- and time-dependent manner, as demonstrated by sub-G0/G1 peaks in DNA content histogram of cell cycle. The cells of G2/M phase by treatment of 0.1 microgram/mL adriamycin had been arrested. G2/M peaks in DNA content was decreased in a dose and time-dependent manner. It had been observed 6 group, 16 genes. The group I contained thioredoxin and cytochrome c oxidase subunit IV gene, group II were p53 and excision repair protein (ERCC-1) gene. Group III was metabolic regulated gene, glucosidase, AMP deaminase isoform L (AMPD2), glutamine synthetase, cholesterol 25-hydroxylase, and steryl-sulfatase precursor. Group IV was cell skeleton constructed gene, heparan sulfate proteoglycan (HSPG2), and microfibrillar-associated protein (MFAP2), group V was oncogene group, v-yes-1 Yamaguchi sarcoma viral oncogene homolog-1 (YES1) and tyrosine kinase ELK1. The other group 6 contained NOD1 protein gene interleukine-1 receptor accessory protein (IL1RAP), pregnancy-specific glycoprotein-11 (PSG11), and pregnancy-specific protein-1a (PSG-1a). CONCLUSION: The present findings indicating that adriamycin was revealed apoptosis in Hela cell. Differential gene expression is related in various metabolism by adriamycin.

Cell Cycle and Gene Expression of Adriamycin-Induced Apoptosis in Human Cervical Carcinoma HeLa cells / 대한산부인과학회잡지

OBJECTIVE: This work was demonstrated the induction of apoptosis in response to adriamycin, we checked the cell cycle of adriamycin-induced apoptosis and to investigate whether differential expression is associated with adriamycin-induced genes in human cervical carcinoma HeLa cells. METHODS: Apoptosis was measured by flow cytometry for cell cycle analysis in Hela cells. Differential expression is associated with adriamycin-induced genes in HeLa cells, it was performed to purifiy the RNA, cDNA probe and hybridization. The various different overexpressed genes were determined by gene array analysis (GDA). Analysis were referenced Incyte Genomics Co. (http://www.ncbi.nlm.nih.gov/). RESULTS: We found that adriamycin was induced apoptosis in a dose- and time-dependent manner, as demonstrated by sub-G0/G1 peaks in DNA content histogram of cell cycle. The cells of G2/M phase by treatment of 0.1 microgram/mL adriamycin had been arrested. G2/M peaks in DNA content was decreased in a dose and time-dependent manner. It had been observed 6 group, 16 genes. The group I contained thioredoxin and cytochrome c oxidase subunit IV gene, group II were p53 and excision repair protein (ERCC-1) gene. Group III was metabolic regulated gene, glucosidase, AMP deaminase isoform L (AMPD2), glutamine synthetase, cholesterol 25-hydroxylase, and steryl-sulfatase precursor. Group IV was cell skeleton constructed gene, heparan sulfate proteoglycan (HSPG2), and microfibrillar-associated protein (MFAP2), group V was oncogene group, v-yes-1 Yamaguchi sarcoma viral oncogene homolog-1 (YES1) and tyrosine kinase ELK1. The other group 6 contained NOD1 protein gene interleukine-1 receptor accessory protein (IL1RAP), pregnancy-specific glycoprotein-11 (PSG11), and pregnancy-specific protein-1a (PSG-1a). CONCLUSION: The present findings indicating that adriamycin was revealed apoptosis in Hela cell. Differential gene expression is related in various metabolism by adriamycin.

The differential expression of apoptosis-associated genes in human gastric cancer MGC803 cells induced by diallyl trisulfide / 中国药理学通报

Aim To investigate the differential expression of apoptosis-associated genes in human gastric cancer MGC803 cells induced by diallyl trisulfide(DATS).Methods Growth inhibition against MGC803 cells was assayed by MTT assay;The apoptosis induced by DATS was assessed by Flow cytometry and fluorescent microscope.The apoptosis-associated gene expression of MGC803 cell treated with DATS was determided by Human Apoptosis Gene Array.Apaf-1 and SODD genes were confirmed by RT-PCR.Results DATS had significant growth inhibitory activity against MGC803 cells,inhibition ratio increased from 11% to 78% at 4,8,12,16 and 24 mg?L-1 for 72 h(P

The altered gene profile of human osteosarcoma cell after knock-down of double functional gene apurinic/apyrimidinic endonuclease / 第三军医大学学报

Objective To explore the molecular mechanism of inhibited malignant phenotype by knock-down of apurinic/apyrimidinic endonuclease(APE1) in human osteosarcoma cell HOS,as well as the possible interacting molecules among APE1.Methods Knock-down of APE1 by synthesized APE1 siRNA in HOS cell was identified first by Western blotting and AP endonuclease assay,then the gene profile was determined with GEArray~(TM) series Human CancerPathwayFinder gene array.Results The specific knock-down of APE1 was found in HOS cell transfected with APE1 siRNA,and its inhibited rate was 91.5%.Six biological pathways of the gene array were involved in HOS cell after knock-down of APE1,but the cell cycle pathway was less influenced.Forty-two out of 96 genes were altered(43.8%),in which only CDK4,FGFR2,KAI1 and NCAM were increased,38 others were decreased.Conclusion The knock-down of APE1 was detected to be involved in all the pathways including cell cycle and DNA repair,apoptosis,signal transduction,adhesion,angiogenesis,invasion and metastasis.These findings are significant to further elucidate APE1 role in tumor genesis and to provide experimental evidence for gene therapy targeting APE1 gene in the future.