ABSTRACT
Objective:To explore the role of BMP7 in inducing the differentiation of BMSCs into neurons in vitro.Methods:BMSCs were isolated and cultured by whole bone marrow adherence method. Adipogenic induction and osteogenic differentiation were used to test the multi-directional differentiation ability of BMSCs. BMSCs were randomly divided into control group and BMP7 groups (25 ng/ml, 50 ng/ml, 75 ng/ml and 100 ng/ml). The effect of BMP7 on the proliferation of rat BMSCs was measured by MTT assay. BMP7 induced morphological changes in rat BMSCS under an inverted phase contrast microscope. The relative expression levels of NF200, SYN1, MAP2 and GFAP mRNA in induced cells were measured by qRT-PCR. Immunofluorescence was used to measure the expression of NSE protein.Results:Rat BMSCs showed lipid droplets in the cytoplasm after adipogenic induction, and oil red O staining was positive; rat BMSCs showed opaque mineral nodules after osteogenic induction, and alizarin red stained was positive. At 2 d, 3 d, 4 d, 5 d, and 6 d after induction, the cell absorbance values of each group were statistically different ( P < 0.05). On the 6th day of induction, the absorbance values of the control group, 25 ng/ml BMP7 group, 50 ng/ml BMP7 group, 75 ng/ml BMP7 group and 100 ng/ml BMP7 group were 0.370±0.003, 0.399±0.003, 0.404±0.003, 0.410±0.003, 0.397±0.001, respectively. Cells absorbance value of 75 ng/ml BMP7 group was significantly higher than the other groups ( P< 0.05). The 75 ng/ml BMP7 group had the most significant changes in cell morphology, similar to neurons in cell morphology. The relative expression of NF200, SYN1, MAP2, and GFAP mRNA of 75 ng/ml BMP7 group (5.47±0.59, 1.48±0.38, 2.86±1.65, 4.41±0.13) was significantly higher than that of the control group ( P< 0.05). The positive rate of NSE immunofluorescence staining in the 75 ng/ml BMP7 group was higher than that in the control group (32.94%±1.62% vs 0). Conclusion:BMP7 has the ability to induce the differentiation of rat BMSCs into neurons in vitro.
ABSTRACT
The formation of osteolytic bone lesions is a key process for osteolytic cancer to metastasize to the bone and is under the control of a set of transcription factors. Recently, the inhibitor of differentiation 1 (Id1) has been linked with angiogenesis, tumorigenesis, metastasis and bone formation. However, the function of Id1 during the process of bone destruction caused by cancer in vivo has not yet been elucidated. We, therefore, examined whether and how Id1 affects the ability of cancer to form osteolytic lesion in vivo. The study used a lentiviral vector overexpressing short hairpin RNA (shRNA) targeting Id1 gene. PC3 cells, a prostate cancer cell line, were transduced with Id1 shRNA or negative control (NC) shRNA before implantation in BALB/c mice. Cells were implanted in a tibial injection model. Tumor formation in bone was monitored by X-ray. The relationship between parathyroid hormone-related protein (PTHrP), an osteolytic factor, and Id1 was analyzed by using immunohistochemistry in tissue sections from osteolytic lesion of the BALB/c mice. Our results showed that Id1 shRNA delivery to PC3 cells by lentivirus caused efficient and stable Id1 gene silencing. In the intratibial model, PC3 cells produced primarily osteolytic lesions in the bone. Eleven of 14 mice in Id1 shRNA group but only 4 of 14 mice in the NC shRNA group developed osteolytic lesions with cortical destruction at 4th week. Mice treated with Id1 shRNA had larger tumor volume in the bone and larger cortical destruction. The expression of PTHrP protein in PC3 cells was not affected by Id1 knockdown in vivo. These results indicate that Id1 may down-regulate the ability of PC3 cells to form osteolytic lesions in vivo and the signal pathway needs to be further investigated.
Subject(s)
Animals , Humans , Male , Mice , Bone Neoplasms , Genetics , Metabolism , Cell Line, Tumor , Gene Silencing , Inhibitor of Differentiation Protein 1 , Genetics , Metabolism , Mice, Inbred BALB C , Osteolysis , Genetics , Metabolism , Pathology , Prostatic Neoplasms , Genetics , Metabolism , PathologyABSTRACT
The formation of osteolytic bone lesions is a key process for osteolytic cancer to metastasize to the bone and is under the control of a set of transcription factors. Recently, the inhibitor of differentiation 1 (Id1) has been linked with angiogenesis, tumorigenesis, metastasis and bone formation. However, the function of Id1 during the process of bone destruction caused by cancer in vivo has not yet been elucidated. We, therefore, examined whether and how Id1 affects the ability of cancer to form osteolytic lesion in vivo. The study used a lentiviral vector overexpressing short hairpin RNA (shRNA) targeting Id1 gene. PC3 cells, a prostate cancer cell line, were transduced with Id1 shRNA or negative control (NC) shRNA before implantation in BALB/c mice. Cells were implanted in a tibial injection model. Tumor formation in bone was monitored by X-ray. The relationship between parathyroid hormone-related protein (PTHrP), an osteolytic factor, and Id1 was analyzed by using immunohistochemistry in tissue sections from osteolytic lesion of the BALB/c mice. Our results showed that Id1 shRNA delivery to PC3 cells by lentivirus caused efficient and stable Id1 gene silencing. In the intratibial model, PC3 cells produced primarily osteolytic lesions in the bone. Eleven of 14 mice in Id1 shRNA group but only 4 of 14 mice in the NC shRNA group developed osteolytic lesions with cortical destruction at 4th week. Mice treated with Id1 shRNA had larger tumor volume in the bone and larger cortical destruction. The expression of PTHrP protein in PC3 cells was not affected by Id1 knockdown in vivo. These results indicate that Id1 may down-regulate the ability of PC3 cells to form osteolytic lesions in vivo and the signal pathway needs to be further investigated.
ABSTRACT
Objective To explore whether or not the cartilage damage in the pathogenesis of pathologic synovial plicae is due to the matrix metalloproteinase. Methods The immunohistochemical method was used to detect the expression and distribution of matrix metalloproteinase-1(MMP-1) and tissue inhibitor of metalloproteinase-1(TIMP-1) in arthroscopically clarified pathologic synovial plicae and normal synovial plicae of the knee joint.Results The positive expression rate of MMP-1 and TIMP-1 had significant differences between the pathologic and normal synovial plicae(?~2=16.014,P=0.000;?~2=4.059,P=0.044).The expression of MMP-1 was positive in the synovial lining cells,monocytes,lymphocytes,endothelial cells,and chondrocytes,but negative in the normal synovial plicae.The TIMP-1 expression was only detected in the synovial lining cells and a small quantity of fibroblast cells.The immunohistochemical analysis revealed a greater number of positive cells and intensity of staining of MMP-1 than TIMP-1.Conclusions The development of pathologic synovial plicae may yield MMP-1 and TIMP-1 with unbalanced distributions,which may be the biological basis of the pathogenesis of cartilage destruction.