OBJECTIVETo observe the influence of Wnt-1 recombinant adenovirus on differentiation tendency of human epidermal stem cells.

METHODSWnt-1 recombinant adenovirus was transduced into hESCs (E group), while normal hESCs were used as control (C) group. The diameter, proliferation,and labeling molecular expression of hESC were determined. The content of MMP-2 and MMP-7 in supernate were also assayed.

RESULTSThere was no obvious difference in diameter of hESC between two groups. The density of hESC in E group was (1.45 +/- 0.09) x 10(5)/mL, which was obviously higher than that in C group [(1.18 +/- 0.10) x 10(5)/mL, P < 0.05]. There were no obvious differences in expression of markers between two groups,including keratin 5 (KS), K6, K7, KS, K14, CD44, carcinoembryonic-like antigen (CEAA), ER, PR (P > 0.05) ,while the expression of K 10 was different among groups [(60 +/- 3)% in E group, 0 in C group], also K18 [(34.3 +/- 2.1)% in E group vs. (13.8 +/- 1.7)% in C group, P < 0.05], and K19 [(17.1 +/- 1.8)% in E group vs. (24.4 +/- 1.5)% in C group, P < 0.05].The contents of MMP-2 and MMP-7 in E group were higher than those in C group (P < 0.01).

CONCLUSIONWnt-1 recombinant adenovirus can induce the differentiation of hESCs to glandular epithelium-like cells.

Adenoviridae ; genetics ; Cell Differentiation ; Cell Line ; Epithelial Cells ; cytology ; virology ; Humans ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 7 ; metabolism ; Stem Cells ; cytology ; Wnt1 Protein ; genetics

To explore the effects of the physiological range of hydrostatic pressure on human bladder smooth muscle cells (HBSMCs) cultured in vitro, we used a hydrostatic compression device designed in our laboratory into the experiments, which were grouped by varied hydrostatic pressure gradients. Cellular morphology was observed with HE staining; cytoskeleton F-actin, cell cycle, both proliferating cell nuclear antigen (PCNA) and matrix metalloproteinase 7 (MMP-7) were detected respectively with immunofluorescence, flow cytometry and RT-PCR. We found that the proliferation, cytoskeleton and cycle distribution of HBSMCs were not obviously different among the groups of different hydrostatic pressure; however, the mRNA expression of MMP-7 exhibited a trend of first increasing and then declining as the pressure gradually rises. Thus the physiological range of hydrostatic pressure may not have significant influence on proliferation, morphology, skeleton, and cell cycle of HBSMCs, but it may have great effect on the expression of MMP-7.
Cells, Cultured ; Humans ; Hydrostatic Pressure ; Matrix Metalloproteinase 7 ; genetics ; metabolism ; Myocytes, Smooth Muscle ; cytology ; physiology ; Proliferating Cell Nuclear Antigen ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Urinary Bladder ; cytology

OBJECTIVETo investigate that the role of Axin in regulating the invasion and migration ability of lymphoma cells and explore the molecular mechanisms.

METHODSThe expressions of Axin, β-catenin, MMP7, and MMP9 were detected in different lymphoma cell lines by RT-PCR and Western blotting. A lymphoma cell line with low Axin expressions was transiently transfected with pCMV5-HA-Axin and pcDNA5-His-β-catenin plasmid, and the expressions of β-catenin, MMP7, and MMP9 mRNA and protein were observed. A lymphoma cell model stably overexpressing Axin was transfected with AXIN-shRNA and β-catenin-shRNA, and the changes in β-catenin, MMP7, and MMP9 cexpressions were observed. The changes in the invasion and migration abilities of this cell model were assessed following Axin knockdown.

RESULTSIn the lymphoma cell lines tested, the Axin expression showed a negative correlation with β-catenin, MMP7, and MMP9 expressions. In Raji cells with a low Axin expression, overexpression of Axin resulted in decreased expressions of β-catenin, MMP7, and MMP9 at the protein levels but not the mRNA levels, and overexpression of β-catenin obviously increased MMP7 and MMP9 mRNA and protein expressions. In the cells with stable Axin overexpression, Axin knockdown caused increased expressions of β-catenin, MMP7, and MMP9 at the protein levels but not the mRNA levels, while β-catenin knockdown caused lowered expressions of MMP7 and MMP9 and suppressed cell invasion and migration.

CONCLUSIONIn lymphoma cells, Axin overexpression can decrease the expression of β-catenin, which in turn decreases the expressions of MMP7 and MMP9 to inhibit the cell invasion and migration.

Axin Protein ; genetics ; metabolism ; Cell Line, Tumor ; Down-Regulation ; Gene Knockdown Techniques ; Humans ; Lymphoma ; genetics ; metabolism ; pathology ; Matrix Metalloproteinase 7 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Neoplasm Invasiveness ; Neoplasm Metastasis ; RNA, Messenger ; RNA, Small Interfering ; Transfection ; beta Catenin ; metabolism

OBJECTIVE: To explore the tissue-specific gene expressions of the peripheral blood and the menstrual blood, and to search some specific factors to establish an effective method for identifying the peripheral blood and the menstrual blood. METHODS: The specific products of the peripheral blood and the menstrual blood were detected by RT-PCR and separated by electrophoretic technology. RESULTS: Beta-spectrin (SPTB) as one specific marker of peripheral blood and 18S rRNA as a kind of the housekeeping gene were expressed in both the peripheral blood and the menstrual blood. However, matrix metalloproteinase 7 (MMP7) as one specific marker of menstrual blood and human beta defensin 1 (HBD1) as one specific marker of vaginal discharge were only found in the menstrual blood. CONCLUSION There are differences of specific gene expressions between the peripheral blood and the menstrual blood. They could be accurately distinguished from each other by using the combination of fluorescence technology and RT-PCR to detect the specific identification of mRNA.
Biomarkers ; Blood/metabolism* ; Female ; Gene Expression ; Gene Expression Profiling ; Humans ; Matrix Metalloproteinase 7/genetics* ; Menstruation/genetics* ; RNA, Messenger/genetics* ; Reverse Transcriptase Polymerase Chain Reaction ; beta-Defensins

Overexpression of HER2 correlates with more aggressive tumors and increased resistance to cancer chemotherapy. However, a functional comparison between the HER2high/HER3 and the HER2low/HER3 dimers on tumor metastasis has not been conducted. Herein we examined the regulation mechanism of heregulin-beta1 (HRG)-induced MMP-1 and -9 expression in breast cancer cell lines. Our results showed that the basal levels of MMP-1 and -9 mRNA and protein expression were increased by HRG treatment. In addition, HRG-induced MMP-1 and -9 expression was significantly decreased by MEK1/2 inhibitor, U0126 but not by phosphatidylinositol 3-kinase (PI-3K) inhibitor, LY294002. To confirm the role of MEK/ERK pathway on HRG-induced MMP-1 and -9 expression, MCF7 cells were transfected with constitutively active adenoviral-MEK (CA-MEK). The level of MMP-1 and -9 expressions was increased by CA-MEK. MMP-1 and -9 mRNA and protein expressions in response to HRG were higher in HER2 overexpressed cells than in vector alone. The phosphorylation of HER2, HER3, ERK, Akt, and JNK were also significantly increased in HER2 overexpressed MCF7 cells compared with vector alone. HRG-induced MMP-1 and -9 expressions were significantly decreased by lapatinib, which inhibits HER1 and HER2 activity, in both vector alone and HER2 overexpressed MCF7 cells. Finally, HRG-induced MMP-1 and MMP-9 expression was decreased by HER3 siRNA overexpression. Taken together, we suggested that HRG-induced MMP-1 and MMP-9 expression is mediated through HER3 dependent pathway and highly expressed HER2 may be associated with more aggressive metastasis than the low expressed HER2 in breast cancer cells.
Breast Neoplasms/enzymology/*genetics/*metabolism ; Butadienes/pharmacology ; Cell Line, Tumor ; Dose-Response Relationship, Drug ; Enzyme Inhibitors/pharmacology ; Female ; Gene Expression ; Gene Expression Regulation, Neoplastic/drug effects ; Humans ; MAP Kinase Signaling System ; MCF-7 Cells ; Matrix Metalloproteinase 1/*genetics/metabolism ; Matrix Metalloproteinase 9/*genetics/metabolism ; Neuregulin-1/*pharmacology ; Nitriles/pharmacology ; Phosphatidylinositol 3-Kinases/metabolism ; Protein Kinase Inhibitors/pharmacology ; Protein Multimerization ; Proto-Oncogene Proteins c-akt/metabolism ; Quinazolines/pharmacology ; Receptor, erbB-2/genetics/*metabolism ; Receptor, erbB-3/*metabolism

Adult ; Aged ; Animals ; Apoptosis ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Female ; Gene Expression ; Humans ; Immunohistochemistry ; In Situ Hybridization ; Liver ; metabolism ; pathology ; Liver Cirrhosis ; metabolism ; pathology ; Liver Neoplasms ; metabolism ; pathology ; Male ; Matrix Metalloproteinase 7 ; genetics ; metabolism ; Mice ; Middle Aged ; Prognosis ; Proto-Oncogene Proteins c-mdm2 ; genetics ; metabolism ; RNA, Messenger ; metabolism

Triptolide, a compound extracted from the traditional Chinese medicine preparation of Tripterygium wilfordii Hook F., has been reported to have anti-inflammatory and anti-cancer activities. However, its effect on ovarian cancer invasion is unknown. We observed that MMP7 and MMP19 expression increased in ovarian cancer tissue. Triptolide treatment inhibited the migration and invasion of ovarian cancer cells SKOV3 and A2780 at the concentration of 15 nM. We also observed that triptolide suppressed MMP7 and MMP19 promoter activity in a dose-dependent manner, down-regulating the expressions of these promoters on mRNA and protein level. Moreover, triptolide enhanced E-cadherin expression in ovarian cancer cells. In vivo, triptolide inhibited tumor formation and metastasis in nude mice, and suppressed MMP7 and MMP19 expression; it also enhanced E-cadherin expression in tumor in a dose-dependent manner. Over expression of MMP7 and MMP19, or suppression of E-cadherin expression partially abolished the inhibitory effect of triptolide on invasion of ovarian cancer cells. To summarize, triptolide significantly inhibited the migration and invasion of ovarian cancer cells by suppression of MMP7 and MMP19 and up-regulation of E-cadherin expression. This study shows that triptolide is a good candidate for the treatment of ovarian cancer and reduction of metastasis.
Animals ; Antineoplastic Agents, Alkylating/*pharmacology ; Cadherins/*genetics/metabolism ; Cell Line, Tumor ; Cell Movement/drug effects ; Cell Proliferation/drug effects ; Cystadenocarcinoma, Serous/*drug therapy/pathology ; Diterpenes/*pharmacology ; Epoxy Compounds/pharmacology ; Female ; Gene Expression Regulation, Enzymologic/drug effects ; Humans ; Matrix Metalloproteinase 7/genetics/*metabolism ; Matrix Metalloproteinases, Secreted/genetics/*metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Invasiveness ; Ovarian Neoplasms/*drug therapy ; Paclitaxel/pharmacology ; Phenanthrenes/*pharmacology ; Promoter Regions, Genetic ; Up-Regulation/drug effects ; Xenograft Model Antitumor Assays