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Biomedical subjects

Yan-qing Ding

Publications and source records attributed to Yan-qing Ding.

At least 19 recordsLinked to original sources

[Effect of silencing epidermal growth factor receptor expression by RNA interference on the growth of nasopharyngeal carcinoma cell 5-8F].

OBJECTIVE: To observe the growth and cell cycle changes of nasopharyngeal carcinoma cell 5-8F in response to silencing of the expression of epidermal growth factor receptor (EGFR) with RNA interference (RNAi), and explore the possible relationships between EGFR and the occurrence, differentiation and progression of nasopharyngeal carcinoma. METHOD: Three EGFR-specific small interfering RNAs (siRNAs) were obtained by in vitro transcription and synthesis and were transiently transfected into 5-8F cells. The EGFR expression levels in the transfected cells were detected by reverse transcription (RT)-PCR and Western blotting, respectively. After EGFR expression silencing, the growth and cell cycle changes of the cells were observed. RESULTS: EGFR mRNA contents and protein levels decreased by approximately 67.5% and 77%, respectively, after RNAi of 5-8F cells, and the cell proliferation decreased and cell cycle arrest at G1 phase occurred in association with EGFR silencing. CONCLUSION: EGFR silencing by RNAi can reduce the proliferation of nasopharyngeal carcinoma cells and induce cell cycle arrest at G1 phase, which sheds light on the possible use of RNAi for further investigation of the pathogenesis and gene therapy of nasopharyngeal carcinoma.

Cell Line, Tumor↗

[Expression and diagnostic application of C4.4A protein in squamous cell carcinoma and adenocarcinoma].

OBJECTIVE: To investigate the diagnostic utility of C4.4A gene expression in discriminating a squamous cell carcinoma (SCC) from an adenocarcinoma by immunohistochemistry. METHODS: Immunohistochemical staining was performed to detect the expression of C4.4A protein in 157 cases of SCC and 177 cases of adenocarcinoma of various organs. RESULTS: Overall, 141 of 157 cases of SCC strongly expressed C4.4A protein. In contrast, only 8 of 177 adenocarcinomas showed partial or scattered cell expression of C4.4A protein. The statistic difference between the two groups was highly significant (chi(2) = 244.93, P = 0.000), and also when the tumors were stratified according to the degree of differentiation (P = 0.000). CONCLUSION: C4.4A protein expression may serve as a valuable tumor marker in discriminating a squamous cell carcinoma from an adenocarcinoma, and therefore, may greatly facilitate the differential diagnosis of an epithelial malignancy.

Adenocarcinoma↗

[Effect of cell surface sialic acid and their linkages on adhesion of mammary carcinoma cells].

OBJECTIVE: To investigate the effect of cell surface sialic acid and its linkage on the cell-cell and cell-matrix adhesion of mammary carcinoma cells MD-MB-435. METHODS: MD-MB-435 cells were sense-transfected with ST6Gal I cDNA or antisense-transfected with part of the ST6Gal I sequence inserted in pcDNA 3.1 vector, with mock transfection with pcDNA3.1 vector as the control. The cell surface alpha2, 6-linked sialylation was determined by fluorescence-activated cell sorting (FACS) using lectin SNA (Sambucus nigra agglutinin specific to alpha2, 6-linked sialic acid on N-linked glycoprotein). A significantly increased alpha2, 6-sialylation subclone in sense-transfectants and a decreased alpha2, 6-sialylation subclone in antisense-transfectants were selected for further examination of cell-cell and cell-matrix (collagen IV) adhesion. The transfectants were also treated with sialidase to compare the capacity of cell adhesion affected by cell surface sialylation. RESULTS: Sense-transfection subclone showed a reduced cell-cell aggregation but enhanced cell-matrix adhesion. In contrast, the antisense-transfection subclone exhibited increased cell-cell aggregation and decreased cell-matrix adhesion. After treatment with sialidase, the cell-matrix adhesion of all the transfectants and the parental MDA-MB-435 cells were significantly reduced to the level of 31%-57% of untreated cells. CONCLUSION: Cell surface sialic acid and alpha2, 6-linked sialylation play an important role in cell-cell and cell-matrix adhesion of mammary carcinoma cell MDA-MB-435.

Antigens, CD↗

[Association of abnormal cyclooxygenase-2 gene expression with colorectal carcinoma metastasis].

OBJECTIVE: To investigate cyclooxygenase-2 (COX-2) expression of in colorectal carcinoma cell lines and tissues and its clinical implications. METHODS: SP immunohistochemistry was used to detect COX-2 protein in SW480 and SW620 cell lines and 50 primary colorectal carcinoma and 50 lymphoma metastasis carcinoma specimens. Real-time PCR was used to detect COX-2 mRNA expression in SW480 and SW620 cell lines. RESULTS: SW480 and SW620 cell lines both highly expressed COX-2. The positive expression levels of COX-2 increased significantly in lymph node metastatic carcinoma in comparison with primary colorectal carcinoma (P<0.05). The expression COX-2 mRNA in SW620 cell line was higher than that of SW480 cell line, showing a mean expression increment of 2.268 folds in SW620 cell line relative to SW480. CONCLUSION: Elevated COX-2 expression may be associated with lymph node metastases of colorectal carcinoma.

Cell Line, Tumor↗

[Primary diffuse large B-cell lymphoma of the heart: a clinicopathological study].

OBJECTIVE: To define the clinicopathological features of primary cardiac large B-cell lymphoma. METHOD: A case of primary cardiac large B-cell lymphoma was studied with conventional histopathological and immunohistochemical staining in combination with literature review. RESULTS: The lesion appeared to originate in the right atrium and involved the venae cavae and the left atrium. Microscopic examination showed diffuse proliferation of large atypical lymphocytes with abundant cytoplasm, vestiealer nuelei, thick nuclear membrane and conspicuous nucleoli. Giant tumor cells scattered in the lesion. The neoplastic cells were positive for CD20 and CD79a. CONCLUSION: Primary cardiac lymphoma is extremely rare, and its pathogenesis remains unclear. With non-specific clinical manifestations, the majority of primary cardiac lymphomas are of B-cell lineage and a bad prognosis.

Aged↗

[ZNF217 gene was detected in ovarian serous cystadenocarcinoma by fluorescence in situ hybridization].

OBJECTIVE: To investigate amplification of zinc finger protein 217 (ZNF217) gene in ovarian serous cystadenocarcinoma and its clinical significance. METHODS: Twenty three specimens of ovarian carcinoma, 10 specimens of ovarian benign tumors and 7 specimens of normal ovaries and two ovarian cancer cell lines, SKOV3 and HO-8910 were examined by fluorescence in situ hybridization (FISH). RESULTS: The amplification of ZNF217 was gained in 12 case of ovarian cancers, there was only 1 case in ovarian benign tumor and not amplication in normal ovary. CONCLUSION: The amplification of ZNF217 is associated with ovarian cancer. Oncogenes ZNF217 maybe play a role in cell differentiation and indicate poorer survival in patients with ovarian cancer.

Adult↗

Selection of choriocarcinoma-associated genes using bioinformatics.

OBJECTIVE: To analyze the data of gene expression profiles of choriocarcinoma and screen for choriocarcinoma- related genes. METHODS: Human cDNA expression microarray containing 4 096 genes was used to study the gene expression profiles in specimens of complete hydatidiform moles (n=3) and choriocarcinomas (n=3), with normal placental villi serving as the control group. The candidate genes with similar expression profiles were identified by hierarchical cluster analysis, and their expressions in normal and neoplastic tissues analyzed by electronic Northern analysis and other bioinformatics methods. Three selected genes were analyzed by beta-actin semiquantitative reverse transcriptase-PCR to confirm the data. RESULTS: A total of 52 coexpressed candidate genes were identified from the gene expression data derived from the choriocarcinoma specimens, of which 19 genes were selected as choriocarcinoma-related genes by further cluster analysis, such as dynamin (L07807), katanin p60 (AF056022), zinc finger protein ZNF184 (U6656), calmodulin (U12022), carboxypeptidase M (BC022276), calcineurin-binding protein cabin 1 (NM_012295) and transducin-like enhancer protein TLE1(M99435). CONCLUSION: The identification of choriocarcinoma-related genes by cluster analysis provides new clues for seeking the key genes associated with the progression and metastasis of choriocarcinoma.

Choriocarcinoma↗

[Analysis of protein expression maps of two metastatic colorectal carcinoma cell lines generated by two-dimensional gel electrophoresis].

OBJECTIVE: To analyze the differentially expressed proteins in metastatic colorectal carcinoma and provide clues for the molecular mechanisms of colorectal cancer metastasis. METHODS: A pair of colorectal carcinoma cell lines SW620 and SW480 with different metastatic potentials from the same parent cell lines were studied. The protein expression maps of the two cell lines were obtained and optimized using two-dimensional gel electrophoresis (2-DE), and the differentially expressed protein spots associated with metastasis were analyzed by Melanie III software. RESULTS: The 2-DE expression maps of the two cell lines were well matched. The resolution of 2-DE was increased after using the nonlinear pH3-10 or overlapping narrow immobilized pH gradient strips, and 11 differentially expressed protein spots were identified after analysis using the software. CONCLUSION: Nonlinear pH3-10 and overlapping narrow immobilized pH gradient strips have higher separation efficiency for protein in 2-DE. The differentially expressed proteins separated will be useful for identifying metastasis-associated proteins of colorectal carcinoma and for establishing 2-DE database of the metastatic colorectal carcinoma cell lines.

Cell Line, Tumor↗

[Apoptotic characteristics of spleen lymphocyte in mice irradiated by lethal dose and its relationship to the expression of Bax and Bcl-XL proteins].

OBJECTIVE: To observe the apoptotic characteristics of mouse spleen lymphocyte after lethal dose gamma-irradiation and its relationship to the expression of Bax and Bcl-XL proteins. METHODS: Two hundred and twenty-five second-grade C57 mice were randomly divided into six groups of 0, 6, 9, 12, 15 and 20 Gy. They were sacrificed by dislocation and samples were taken on 1-28 days after whole body single gamma-irradiation. Lymphocyte apoptosis and necrosis were analyzed by TdT-mediated dUTP nick end labeling (TUNEL) and flow cytometry (FCM) techniques. The expression of Bax and Bcl-XL proteins were estimated by immunohistochemical method. RESULTS: (1) The number of peripheral white blood cells of mice increased temporarily at 6 hours after radiation, thereafter, began to decrease rapidly, which reached the minimum on day 7 and recovered normal level basically one month after 6 Gy gamma-irradiation. (2) Apoptotic rate of spleen lymphocytes increased significantly, peaking at 6 hours after radiation, which was found to have a dose-response relationship during 6-24 hours after < or =12 Gy irradiation, but decreased after > or =15 Gy irradiation. (3) It was confirmed by FCM that the apoptotic rate of spleen lymphocytes increased along with the elevation of radiation dose. However, the apoptotic rate began to decrease and the necrotic rate rose distinctively after > or =15 Gy irradiation. The analysis of DNA gel electrophoresis supported above-mentioned results. (4) The expression of Bax protein in spleen lymphocyte enhanced at 6 hours after 6 Gy gamma-irradiation and peaked by 12 Gy-irradiation, showing a dose-dependent pattern, but which was not be found after > or =15 Gy gamma-irradiation. On the other hand, the expression of Bcl-XL protein reduced persistently with the increase of radiation dose, and also presented a better dose-dependent effect after < or =12 Gy irradiation. CONCLUSION: After 6-12 Gy gamma-irradiation, the apoptosis is the major death way of spleen lymphocyte, while both necrosis and apoptosis are important death pathways after > or =15 Gy irradiation. Pro-apoptotic Bax and anti-apoptotic Bcl-XL play an important role in the apoptotic regulation of spleen lymphocytes induced by lethal dose radiation.

Animals↗

[Screening of differentially expressed genes in colorectal cancer using human whole genomic oligonucleotide microarrays].

OBJECTIVE: To screen the differentially expressed genes in human colorectal cancer (CRC) tissue. METHODS: Affymetrix oligonucleotide microarrays HG-U133 representing 32,264 human genes including 19,308 known genes and 12,956 expressed sequence tags (ESTs) were used to detect the gene expressions of CRC tissue paired with normal mucosa tissue. The microarray findings were confirmed by real-time quantitative reverse transcriptase-polymerase chain reaction (FQ-PCR). The gene expression profiles were analyzed by intersection and complement, rank sum test and t test. RESULTS: Totally 3,125 genes and ESTs expressed differentially were detected in normal and cancer tissues, consisting of 974 up-regulated and 2,151 down-regulated genes with 247 ESTs present in CRC tissue and absent in normal mucosa and 162 ESTs absent in CRC tissue but present in normal mucosa. A percent of 80.1% of the differentially expressed genes were not reported in the literatures. CONCLUSION: The strategy of data mining provides a foundation for filtering molecular markers and interpreting molecular carcinogenesis of CRC.

Colorectal Neoplasms↗

[Differential proteomic analysis of human colorectal carcinoma cell lines SW620 and SW480 with different metastatic potentials].

OBJECTIVE: To investigate differential protein expression profiles of human colorectal carcinoma cell lines with different metastatic potentials and screen metastasis-associated proteins for analyzing the relationship between metastasis-associated proteins and the tumorigenesis, progression and metastasis of colorectal carcinomas. METHODS: With two-dimensional electrophoresis (2-DE) and matrix assisted laser desorption/ionization-time of flight-mass spectrometry, we analyzed the differentially expressed proteins in two human colorectal carcinoma cell lines SW620 and SW480 with high and low metastatic potentials, and screened for proteins associated with colorectal carcinoma metastasis. RESULTS: Image analysis software Malenie III demonstrated good match and reproducibility of the 2-DE maps obtained from 3 independent experiments, with 1316+/-62 spots detected for SW620 cells and 1332+/-74 spots for SW480 cells with the average matching rate of 82% and 80%, respectively. The spots distributed in the greatest density at the isoelectric points of 4-7 and relative molecular mass weight of 20,000-70,000. Twenty-five distinctly different protein spots (14 spots for SW620 and 11 for SW480) in-gel digested by TPCK trypsin and 23 peptide mass fingerprint maps were obtained by mass spectrometry. Three highly matched proteins and 14 preliminarily matched proteins or fragments were obtained by analysis with Mascot software in the NCBInr database. Some of these differentially expressed proteins were related to gene transcription, cell cycle, signal transduction, cell apoptosis, etc, with possible involvement of cell differentiation, proliferation, invasion, adhesion, and metastasis of colorectal carcinoma. CONCLUSION: The 2-DE protein expression profile of SW620 cells with high metastatic potential displays obvious difference from that of SW480 cells with low metastatic potential, and a variety of proteins can be involved in the metastasis of colorectal carcinoma.

Cell Line, Tumor↗

[Tiam1 expression correlates with the biological behavior of human colorectal carcinoma cells].

OBJECTIVE: To investigate whether the Tiam1 gene expression enhances the invasive and metastatic capabilities of colorectal carcinoma cells. METHODS: Endogenous expression of Tiam1 in five colorectal carcinoma cell lines was investigated by RT-PCR. Tiam1/C1199HA cDNA was transfected into HT29, a colorectal carcinoma cell line without endogenous Tiam1 expression. RNA and protein expression of Tiam1 gene in the transfectants were detected by RT-PCR, immunohistochemistry and Western blot respectively. The biological behaviors of the transfectants were investigated by MTT and in-vitro invasion assays. RESULTS: Tiam1 gene was highly expressed in LoVo and SW620 cells. Low level expression was seen in HCT116 and SW480 and no expression was found in HT29. Transfection of Tiam1 significantly increased the proliferation of HT29 cells along with markedly enhanced in-vitro invasion and metastasis. CONCLUSIONS: Tiam1 gene plays an important role in the invasion and metastasis of colorectal carcinoma. It may be a useful marker for metastasis of colorectal carcinoma.

Biomarkers, Tumor↗

[Hepatosplenic gammadeltaT-cell lymphoma: a clinicopathological study].

OBJECTIVE: To explore the clinicopathologic features and immunophenotype of hepatosplenic gammadeltaT-cell lymphoma. METHODS: A case of hepatosplenic gammadeltaT-cell lymphoma was studied with conventional histopathological and immunohistochemical staining in combination of literature review. RESULTS: Diffuse hepatic and splenic enlargement was found in this case. Microscopically, the liver and spleen showed marked sinusoidal infiltration. The cells of hepatosplenic T-cell lymphoma were homogeneous, medium in size with pale cytoplasm, and the nuclear chromatin was loosely condensed with small inconspicuous nucleoli. The neoplastic cells were positive for CD45RO, CD3 and TIA-1, but negative for CD57, CD20 and CD79a. CONCLUSION: Hepatosplenic gammadeltaT-cell lymphoma is a rare form of peripheral T-cell lymphoma and has usually poor prognosis, which should be differentiated from other lymphomas or leukemia.

Adult↗

Establishment of transgenic mouse model carring the promoter of plunc sequence.

OBJECTIVE: To construct the vector using specific nasopharynx tissue promoter plunc to establish transgeneic mouse model. METHODS: Plunc-EGFP plasmid was digested with XhoI, the purified linearized DNA fragments were recovered by gel extraction and diluted to the final concentration of 4 microg/ml, before introduced into fertilized one-cell mouse eggs by pronuclear microinjection. RESULTS: Thirteen founder mice were obtained, 12 of which were positive for the integrated EGFP gene as detected by PCR, and 3 were positive shown by Southern blotting. CONCLUSION: Specific nasopharynx tissue promoter plunc can effectively induce exogenous gene integration into the mouse genome.

Animals↗

[Changes of biological characteristics of colon carcinoma cell line - LoVo induced by Kai1/CD82 transfection].

OBJECTIVE: To study the influence of Kai1/CD82 transfection on the growth, adherence, separation and invasion potential of LoVo colon carcinoma cell line. METHODS: Kai1/CD82 cDNA was transfected into LoVo cells, and a stable expressing clone was established. In vitro methodology was used to obtain the growth curve and also to detect the adherence, separation and invasion potential of the transfected LoVo cells, in comparison with those of control cells without transfection. RESULTS: Compared with the control, no change was observed in the growth pattern of transfected LoVo cells. The numbers of adherent cells in the two groups were 0.08, 0.63, 0.83, 0.91 (x 10(5)) for the transfected cells and 0.04, 0.48, 0.71, 0.82 (x 10(5)) for the control cells respectively after 10, 20, 30, 40 minutes culture with shaking. The difference at 20, 30 and 40 minutes was statistically significant (P < 0.05). The separation rates of each group were 13%, 20%, 53% for the transfected cells and 11%, 28%, 60% for the control cells, respectively after 5, 10, 15 minutes culture with shaking. The difference at 10 and 15 minutes was statistically significant (P < 0.05). The aggregation rate of the transfected cells was higher than that of the control cells after culture with mild shaking for 5 hours (64.8% vs. 58.6%, P < 0.05). After co-incubation with endothelium cells ECV304, the number of invading cells decreased more in the transfected cells than that in the control cells (6.33/field and 17.67/field, P < 0.05). CONCLUSION: Transfection expression of Kail/CD82 into LoVo cell line results in an increase of cell adherence and aggregation, but a diminished capability of separation and invasion, suggesting that the expression of Kai1/CD82 gene may inhibit the metastatic potential of colon carcinoma.

Antigens, CD↗

[Differences in gene expression profile between high and low metastatic human colorectal cancer cell lines].

OBJECTIVE: To analyze the gene expression profiles in colorectal cancer cell lines including high metastatic LoVo and SW620 cell lines and low metastatic SW480 and LS174T cells lines using cDNA microarray for tumor metastasis-associated gene and to screen specific colorectal cancer metastasis-associated genes. METHODS: The cDNAs reversely transcribed from the mRNA derived respectively from the 4 cell lines and normal colorectal tissues were labeled with cy3 and cy5 prior to hybridization with the microarray containing 399 genes. The acquired image was analyzed by software. RESULTS: There were significant differences in gene expression profiles between high- and low- metastatic cell lines, and 44 up-regulated genes and 60 down-regulated genes were found to have statistical significance. CONCLUSIONS: The complex alterations in gene expression underlie the development of metastasis of colorectal cancer. The genes that are expressed differentially in high- and low- metastatic cell lines are probably related to the high metastatic potentials.

Cell Line, Tumor↗

[Differential screening of novel human metastasis-associated genes of colorectal carcinoma].

OBJECTIVE: To clone novel metastasis-associated genes of colorectal carcinoma and provide clues for the molecular mechanisms of metastasis of colorectal carcinoma. METHODS: By suppression subtractive hybridization (SSH), a pair of colorectal carcinoma cell lines with different metastatic potentials derived from the same parental cell line was used to construct two subtractive cDNA library specific for metastasis-accelerating genes or metastasis-suppressor genes. Partial positive clones in the two libraries were selected randomly and differentially screened, and the differentially expressed gene fragments obtained were sequenced and analyzed with BLAST software. The mRNA expressions of several new genes were confirmed by Northern blot analysis. RESULTS: Two subtractive cDNA libraries had 235 and 232 white clones respectively and more than 90% of the white clones had the inserts. About 200 positive clones selected randomly were differentially screened, and 29 differentially expressed genes were obtained. Fifteen unknown genes were found by sequence and BLAST analysis and had been collected by the GenBank dbEST database with the entry number of CD485499 to CD485513, among which 6 novel genes were located in the fifth chromosome. CONCLUSION: SSH is a reliable strategy for screening novel genes differentially expressed in metastasis of colorectal carcinoma. The fifth chromosome may harbor many new genes related with metastasis of colorectal carcinoma, and identified new genes can be cloned for their full length for further study of their functions.

Base Sequence↗