As placenta barrier controls material exchanges between mother and fetus during pregnancy, studies on the placental transport mechanism of drugs is of great importance to analyze the efficacy, safety and toxicity of drugs. BeWo cells are derived from human choriocarcinomas, which can form monolayer trophoblast cells to offers an efficient placenta barrier in vitro for estimating nutrient and drug intake, efflux and transport. This essay focuses on the establishment and characteristics of BeWo cell model, and the advance on studies on nutrient and drug intake, efflux and transport mechanism by applying the model.
Biological Transport ; Choriocarcinoma ; metabolism ; Female ; Humans ; Placenta ; metabolism ; Pregnancy ; Tumor Cells, Cultured ; metabolism ; Uterine Neoplasms ; metabolism

OBJECTIVE: To observe the expression profile of heat shock proteins (HSPs) including HSP70, inducible HSP90 (HSP86) and aB-crystallin in cells and tissues of lung adenocarcinoma. METHODS: Western blotting and reverse transcriptional-polymerase chain reaction (RT-PCR) were performed to detect the expression of HSP70, HSP86 and aB-crystallin both in the protein and mRNA level respectively. RESULTS: Compared with normal lung tissue and human bronchial epithelium (HBE) cells, RT-PCR and Western blotting showed that the expression of HSP70, HSP86 and alphaB crystallin increased significantly in both the mRNA and protein level in the cancer tissue and A549 human lung adenocarcinoma cells. Among the 3 sub-families of HSPs, the expression of HSP70 mRNA and protein increased most in both the lung tissue of cancer and A549 human adenocarcinoma cell lines. CONCLUSION The expression of HSPs is higher in the lung adenocarcinoma and A549 cells than that in the normal lung tissues and HBE cells. Among the HSP family, HSP70 is the most up-regulated member in the tissue and cells of lung adenocarcinoma.
Adenocarcinoma ; metabolism ; Adenocarcinoma of Lung ; Cells, Cultured ; Epithelial Cells ; cytology ; metabolism ; Heat-Shock Proteins ; metabolism ; Humans ; Lung ; cytology ; metabolism ; Lung Neoplasms ; metabolism ; Tumor Cells, Cultured

OBJECTIVESTo investigate the androgen receptor (AR) isoforms expression in human benign and malignant prostatic tissues and LNCaP cells.

METHODSUsing high resolution isoelectric focusing (IEF), the different expression of AR isoforms were demosntrated in human benign and malignant prostatic tissues and LNCaP cells.

RESULTSData were obtained from 41 AR-positive BPH, three prostatic cancer specimens, and LNCaP cells. From these materials, three types of AR isoforms were detected with pI values at 6.5, 6.0 and 5.3. In the case of BPH tissues, 15 (36.5%) specimens expressed all the three types of isoforms at pI 6.5, 6.0 and 5.3, and 10 (24.4%) samples contained isoforms at pI 6.5 and 5.3, five (12.2%) samples indicated isoforms at pI 6.5 and 6.0, four (9.8%) showed the isoforms at pI 6.0 and 5.3. Of all the 41 specimens, two (4.9%) and two (4.9%) as well as three (7.3%) denoted the isoforme at pI 6.5, 6.0 and 5.3 respectively. As for three prostatic cancer specimens, one sample showed all the three types of AR isoforms at pI 6.5, 6.0, 5.3, but another specimen expressed at pI 6.5 and 6.0, and only one failed to indicate any types of isoforms. LNCaP cells expressed all three types of AR isoforms at pI 6.5, 6.0 and 5.3. Binding of 3H-dihydrotestosterone to these three types of isoforms was inhibited by the addition of 100-fold excess of DHT and testosterone. No effect of progesterone, oestradiol and diethylstilboestrol on tritiated hormone binding was observed.

CONCLUSIONSThe expression of AR isoforms is different among various patients and different between BPH and LNCaP cells, though no clear explanation could be induced for this. These results suggest the possibility of explaining effective hormonal therapy to prostatic disease in the future.

Humans ; Male ; Prostate ; metabolism ; Protein Isoforms ; metabolism ; Receptors, Androgen ; metabolism ; Tumor Cells, Cultured

This study was aimed to investigate the expression of beta-catenin in leukemic cell lines and its relationship with pathogenesis of leukemia, semi-quantitative RT-PCR and Western blot were performed to detect the expression of beta-catenin in a panel of 15 human hematopoietic cell lines (U937, KG1a, Jurkat, K562, Namalwa, HEL, HUT78, Raji, Daudi, CEM, LCL-H, HL-60, NB4, J6-1, Ramos). Immunocytochemistry was performed in some of these cell lines to detect the location of beta-catenin. The results showed that the beta-catenin gene was widely expressed in most leukemic cell lines in various degree, the high expression of beta-catenin was found is U937, KG1a, Jurkat, K562 and Namalwa cells, middle expression of beta-catenin was observed in HEL, HUT78, Raji, Daudi and CEM cells, lower expression of beta-catenin was observed in LCL-H, HL-60, NB4, J6-1 and Ramos cells. The expression level of beta-catenin protein was identical to the expression level of beta-catenin mRNA. The expression of beta-catenin could be found in nuclei of all cells mentioned above, but their levels were different between them. Abundant beta-catenin also could be observed in nuclei of some leukemic cells by immunocytochemistry. It is concluded that overexpression of beta-catenin in leukemia cells, as a key mediator of Wnt signaling transduction pathway, indicates that the Wnt signaling transduction pathway may be aberrantly activated in leukemia.
Humans ; Leukemia ; metabolism ; pathology ; RNA, Messenger ; metabolism ; Signal Transduction ; Tumor Cells, Cultured ; beta Catenin ; metabolism

Androgen receptor(AR) plays an important role in modulating the effects of androgen on target cells. It is well known that AR is mainly existed in testis. This paper reviewed the distribution of AR and its mRNA in prostate, epididymis, skin of penis, and some other tissues in non-genital system and tumors, such as the skin of scalp, hippocampus, fat, gastric cancer, cancer of larynx, and so on. Besides, this paper also reviewed the detection methods to AR, and further investigated the function of androgen.
Androgens ; metabolism ; Hippocampus ; metabolism ; Humans ; Male ; Penis ; metabolism ; Prostate ; metabolism ; Receptors, Androgen ; metabolism ; Testis ; metabolism ; Tissue Distribution ; Tumor Cells, Cultured

The high expression of tissue factor (TF) is related to the coagulation disorder in acute leukemia. TF in blood circulation is mainly expressed in cells, microparticles (MP) and alternatively spliced human tissue factor (asHTF). To elucidate the role of TF in the coagulation disorder of acute myeloid leukemia (AML), RT-PCR was performed on 6 common AML cell lines NB4, HL-60, Kasumi-1, U937, K562 and THP-1. The results showed that only NB4 and U937 cells expressed baseline full-length TF and asHTF which were proved by sequencing. The flow cytometric detection, TF activity and TF antigen tests in NB4 and U937 cells revealed that the asHTF was expressed in trace amount and almost had no activity, while the TF antigen and activity in microparticles were significantly higher than that in asHTF. It is concluded that asHTF may play an unimportant role in the coagulation disorder of AML. Microparticle associated tissue factor (MP-TF) is the predominant source of TF activity released from AML cells.
Alternative Splicing ; HL-60 Cells ; Humans ; Leukemia, Promyelocytic, Acute ; genetics ; metabolism ; Thromboplastin ; genetics ; metabolism ; Tumor Cells, Cultured ; U937 Cells

AIMTo study the influence of acetylcholine (ACh) on nicotinic receptor(N receptor) beta-subunit of the gastric epithelia and the gastric adenocarcinoma cells, and the difference of both cells.

METHODSImmunohistochemistry method was used to examine the number, number density and surface density of N receptor beta-subunit in both cells cultured in vitro.

RESULTSThe number and number density of N receptor beta-subunit in the gastric adenocarcinoma cells were much more than that in the gastric epithelia (P < 0.05). But surface density of N receptor beta-subunit in the gastric adenocarcinoma cells were lower than that in the gastric epithelia (P < 0.05). ACh at 10(6) mol/L could increase the number, number density and surface density of N receptor beta-subunit in the gastric epithelia (P < 0.01). The increase effect could not be blocked by atropine. ACh had no effect on N receptor beta-subunit in the gastric adenocarcinoma cells.

CONCLUSIONACh at low concentration initiates N receptor desensitization in the gastric epithelia. ACh has no effect on sensitivity of N receptor beta-subunit in the gastric adenocarcinoma cells.

Acetylcholine ; pharmacology ; Adenocarcinoma ; metabolism ; Cells, Cultured ; Epithelial Cells ; drug effects ; Gastric Mucosa ; cytology ; Humans ; Receptors, Nicotinic ; metabolism ; Stomach Neoplasms ; metabolism ; Tumor Cells, Cultured

BACKGROUNDProstate stromal cells are known to regulate epithelial growth as well as support and maintain epithelial function. However, how stromal cells regulate epithelial cells and what differences among various histological/pathological prostate stromal cells in prostate cancer progression still remain unclear. This study aimed to investigate the different phenotypes of human various histological/pathological prostate stromal cells, and their role in tumor promotion.

METHODSThe different phenotypes of the human normal prostatic peripheral zonal primary stromal cells (NPPF), transitional zonal primary stromal cells (NPTF), and prostate cancer associated primary stromal cells (CAF) were examined with growth curves and Annexin V-fluorescein isothiocyanate (FITC) assay. The different effects on prostate cancer cell line C4-2B by NPPF, NPTF, and CAF were examined with MTT assay and Annexin V-FITC assay. The gene expression of different histological/pathological prostate stromal cells was profiled by microarray and hierarchical cluster analysis.

RESULTSThe growth rate of NPPF, NPTF and CAF gradually increased, followed by decreasing apoptosis. In vitro stromal-C4-2B cell line co-culture models, the proliferation and apoptosis of C4-2B cell line were differently affected by human various histological/pathological prostate stromal cells. CAF showed the most powerful effect to C4-2B cell line, as opposed to a weakest effect of NPTF. Microarray and hierarchical cluster analysis showed that the differentially expressed genes of CAF and NPPF were less than NPPF and NPTF, or CAF and NPTF. This was consistent with clinical observations that prostate cancer mostly derived from the peripheral zone and does not usually occur in the transitional zone.

CONCLUSIONNPPF, NPTF and CAF possess extremely different biological characteristics and gene expression, which may play an important role in genesis and development of prostate cancer.

Adult ; Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Cells, Cultured ; Cluster Analysis ; Flow Cytometry ; Humans ; Immunohistochemistry ; Male ; Prostate ; cytology ; Prostatic Neoplasms ; pathology ; Stromal Cells ; cytology ; metabolism ; Tumor Cells, Cultured

As CD40 transduces activation signals involved in inflammatory and immune disorders, we explored the expression and response to CD40 engagement in human glioma cell lines in this study. The CD40 expression in BT-325 and U251 cells was flow cytometrically detected. The cells were incubated with srhCD40L for 72 h to assess its effects on cell growth in vitro. TNF-α expression was quantified by real-time PCR, and protein expression was analyzed by ELISA. The I-κb mRNA was detected by RT-PCR. I-κB expression decreased after stimulation with 1 μg/mL srhCD40L, but it was upregulated after the cells were pretreated with CD40 antibody. srhCD40L significantly inhibited the proliferation of the CD40+ human glioma cells. The stimulation of CD40+ glioma cells with soluble CD40L (CD154) up-regulated the expression of TNF-α at both mRNA and protein levels. We are led to conclude that CD40L/CD40 could inhibit human glioma cells through I-κb signaling pathway. Interferon-γ can augment CD40 expression and the inhibitory effect of CD40 ligand on cell growth in vitro. These results suggest that srhCD40L may benefit the therapy strategy of glioma.
CD40 Antigens ; metabolism ; CD40 Ligand ; metabolism ; Cell Proliferation ; Cells, Cultured ; Glioma ; genetics ; metabolism ; pathology ; Humans ; NF-kappa B ; metabolism ; Signal Transduction ; physiology ; Tumor Cells, Cultured

This study was purposed to investigate the connexin 43 (Cx43) expression level in acute leukemia bone marrow stromal cells (ABMSCs) and normal bone marrow stromal cells (NBMSCs), and to explore the difference in communicating functions between these cells. The Cx43 expression levels of ABMSCs and NBMSCs were detected by using immunohistochemistry and computer gray scale assay, and the difference of gap junction intercellular communication (GJIC) was examined through dry transfer technique. The results showed that expression level of Cx43 in ABMSCs was lower than that in NBMSCs and its function of GJIC in ABMSCs was also weaker than that in NBMSCs. It is concluded that cell-cell communication function is lowered in ABMSCs.
Acute Disease ; Bone Marrow Cells ; cytology ; metabolism ; pathology ; Cell Communication ; physiology ; Connexin 43 ; metabolism ; Gap Junctions ; metabolism ; Humans ; Leukemia ; metabolism ; Stromal Cells ; cytology ; metabolism ; Tumor Cells, Cultured