ABSTRACT
<p><b>OBJECTIVE</b>To explore cell culture techniques for amplification of oval cells with preservation simultaneously of the stem cell characteristics.</p><p><b>METHODS</b>Oval cell line OC3 was cultured in RPMI 1640 supplemented with 15% fetal bovine serum and 20 µg/L EGF. Cells were harvested every 5 passages and were examined with biomarkers including OV-6, c-kit, gamma-glutamyl transpeptidase, placental form of glutathione-S-transferase (GST-P), pyruvate kinase M₂, pyruvate kinase L and albumin using techniques including RT-PCR, immunocytochemistry, and enzymo-cytochemistry.</p><p><b>RESULTS</b>OC3 cell lines could be amplified abundantly in-vitro associating with expression of infant liver cell markers at various level, including OV-6, c-kit, gamma-glutamyl transpeptidase, GST-P, pyruvate kinase M₂, but no expression of mature hepatocyte markers detected including pyruvate kinase L and albumin.</p><p><b>CONCLUSIONS</b>Amplification of OC3 cells with preservation of the stem cell phenotype and high proliferation index can be achieved up to the 79(th) passages by culturing in RPMI 1640 supplemented with 15% fetal bovine serum and 20 µg/L EGF.</p>
Subject(s)
Animals , Rats , Antigens, Differentiation , Metabolism , Cell Culture Techniques , Cell Differentiation , Cell Line , Culture Media , Glutathione Transferase , Metabolism , Hepatocytes , Cell Biology , Metabolism , Liver , Cell Biology , Phenotype , Proto-Oncogene Proteins c-kit , Metabolism , Pyruvate Kinase , Metabolism , Rats, Sprague-Dawley , Stem Cells , Cell Biology , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To characterize the biologic featrues of hepatic oval cells and their protein expression profiles during induced differentiation in vitro.</p><p><b>METHODS</b>Rat hepatic oval cells were treated with epidermal growth factor (EGF) and hepatocyte growth factor (HGF) in vitro, followed by morphological and molecular marker assessment by electromicroscopy, immunocytochemistry, RT-PCR and protein expression chip technology.</p><p><b>RESULTS</b>Ten weeks after induction, the levels of GST-P mRNA and M2-PK mRNA were significantly reduced, whereas those of ALB and CK18 were elevated. Significant variations of expression was seen in 8 protein species during the course of the induced differentiation.</p><p><b>CONCLUSION</b>Combined EGF and HGF treatment in vitro induces cell differentiation of hepatic oval cells, a process in which 8 protein species may play some regulatory roles.</p>
Subject(s)
Animals , Rats , Albumins , Metabolism , Cell Differentiation , Epidermal Growth Factor , Pharmacology , Glutathione Transferase , Genetics , Hepatocyte Growth Factor , Pharmacology , Hepatocytes , Cell Biology , Metabolism , Immunohistochemistry , Keratin-18 , Metabolism , Protein Array Analysis , Pyruvate Kinase , Genetics , RNA, Messenger , Metabolism , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
<p><b>OBJECTIVE</b>To study the expression of membrane MICA (mMICA), soluble MICA (sMICA) and NKG2D receptor in cases of osteosarcoma and to analyze its clinical significance.</p><p><b>METHODS</b>Expression of mMICA in osteosarcoma tissue of 43 cases was detected with immunohistochemistry. Expression of NKG2D in peripheral blood lymphocytes of 16 cases was analyzed by flow cytometry. Serum level of soluble MICA (sMICA) was measured by ELISA.</p><p><b>RESULTS</b>mMICA was widely expressed in osteosarcoma tissue (37/43). Expression of NKG2D in peripheral blood lymphocytes was significantly decreased. High levels of mMICA and NKG2D expression were associated with better differentiation and earlier tumor stage of osteosarcoma (P < 0.05). A significant increase in serum level of sMICA was demonstrated in patients with metastasis and advanced tumor.</p><p><b>CONCLUSIONS</b>The mMICA expression in tumor tissue, NKG2D expression in peripheral lymphocytes and serum sMICA level correlate with the differentiation and stage of osteosarcoma. These parameters may thus represent potential diagnostic and prognostic markers in patients with osteosarcoma. Manipulation of the MICA-NKG2D pathway may become a target of immunotherapy for osteosarcoma.</p>
Subject(s)
Humans , Bone Neoplasms , Blood , Metabolism , Pathology , Cell Differentiation , Enzyme-Linked Immunosorbent Assay , Flow Cytometry , Histocompatibility Antigens Class I , Blood , Metabolism , Immunohistochemistry , Lymphocytes , Allergy and Immunology , NK Cell Lectin-Like Receptor Subfamily K , Metabolism , Neoplasm Staging , Osteosarcoma , Blood , Metabolism , PathologyABSTRACT
<p><b>OBJECTIVE</b>To investigate the expression of interleukin-8 (IL-8) and its prognostic significance in breast cancer.</p><p><b>METHODS</b>Expression of IL-8 in 113 breast cancers, 19 breast benign tumors and 20 breast normal tissues was examined by tissue microarray using immunohistochemistry, and the association of IL-8 expression with patient's clinico-pathological characteristics and prognosis was further analyzed.</p><p><b>RESULTS</b>The positive rate of IL-8 expression in breast cancer was 27.4%, which was significantly higher than that in benign tumor and normal tissue of breast (P = 0.002). IL-8 expression related to histological type (P = 0.040) and lymph node status (P = 0.021). The expression of IL-8 was observed to correlate negatively with ER and PR status (P = 0.015 and P = 0.034), and correlate positively with C-erbB-2 status (P = 0.002). In addition, Kaplan-Meier curves of disease-free survival analysis showed a significant difference between IL-8 positive groups and negative group (P = 0.031).</p><p><b>CONCLUSIONS</b>IL-8 might be a poor prognostic factor for human breast cancer, and also might be a novel molecular marker to predicate the occurrence and progression of breast cancer.</p>