Despite significant advances in monoclonal antibodies (Mabs) technology over the last two decades, technology to develop natural occurring and fully human therapeutic Mabs from patients without known antigens is still...Despite significant advances in monoclonal antibodies (Mabs) technology over the last two decades, technology to develop natural occurring and fully human therapeutic Mabs from patients without known antigens is still in its infancy. Human myeloma cell lines have been very difficult to derive. Attempts in numerous laboratories to use those cells for the production of human monoclonal antibodies have failed despite early reports. A human myeloma cell line that is suitable for the generation of human monoclonal antibodies (Proceedings of the National Academy of Sciences USA 98:1799-1804) was developed by Dr. Karpas. Since 1975 researchers across the world have been trying to replicate the process with human cells.展开更多
AIM: To understand which and how different miRNAs are implicated in the process of hepatic stellate cell (HSC) activation. METHODS: We used microarrays to examine the differential expression of miRNAs during in vitro ...AIM: To understand which and how different miRNAs are implicated in the process of hepatic stellate cell (HSC) activation. METHODS: We used microarrays to examine the differential expression of miRNAs during in vitro activation of primary HSCs (pHSCs). The transcriptome changes upon stable transfection of rno-miR-146a into an HSC cell line were studied using cDNA microarrays. Selected differentially regulated miRNAs were investigated by quantitative real-time polymerase chain reaction during in vivo HSC activation. The effect of miRNA mimics and inhibitor on the in vitro activation of pHSCs was also evaluated.RESULTS: We found that 16 miRNAs were upregulated and 26 were downregulated significantly in 10-d in vitro activated pHSCs in comparison to quiescent pHSCs. Overexpression of rno-miR-146a was characterized by marked upregulation of tissue inhibitor of metalloproteinase-3, which is implicated in the regulation of tumor necrosis factor-α activity. Differences in the regulation of selected miRNAs were observed comparing in vitro and in vivo HSC activation. Treatment with miR-26a and 29a mimics, and miR-214 inhibitor during in vitro activation of pHSCs induced significant downregulation of collagen type Ⅰ transcription. CONCLUSION: Our results emphasize the different regulation of miRNAs in in vitro and in vivo activated pHSCs. We also showed that miR-26a, 29a and 214 are involved in the regulation of collagen type I mRNA.展开更多
文摘Despite significant advances in monoclonal antibodies (Mabs) technology over the last two decades, technology to develop natural occurring and fully human therapeutic Mabs from patients without known antigens is still in its infancy. Human myeloma cell lines have been very difficult to derive. Attempts in numerous laboratories to use those cells for the production of human monoclonal antibodies have failed despite early reports. A human myeloma cell line that is suitable for the generation of human monoclonal antibodies (Proceedings of the National Academy of Sciences USA 98:1799-1804) was developed by Dr. Karpas. Since 1975 researchers across the world have been trying to replicate the process with human cells.
基金Supported by Institute of Bioengineering and Nanotechnology (Biomedical Research Council, Agency for Science, Technology and Research, Singapore)
文摘AIM: To understand which and how different miRNAs are implicated in the process of hepatic stellate cell (HSC) activation. METHODS: We used microarrays to examine the differential expression of miRNAs during in vitro activation of primary HSCs (pHSCs). The transcriptome changes upon stable transfection of rno-miR-146a into an HSC cell line were studied using cDNA microarrays. Selected differentially regulated miRNAs were investigated by quantitative real-time polymerase chain reaction during in vivo HSC activation. The effect of miRNA mimics and inhibitor on the in vitro activation of pHSCs was also evaluated.RESULTS: We found that 16 miRNAs were upregulated and 26 were downregulated significantly in 10-d in vitro activated pHSCs in comparison to quiescent pHSCs. Overexpression of rno-miR-146a was characterized by marked upregulation of tissue inhibitor of metalloproteinase-3, which is implicated in the regulation of tumor necrosis factor-α activity. Differences in the regulation of selected miRNAs were observed comparing in vitro and in vivo HSC activation. Treatment with miR-26a and 29a mimics, and miR-214 inhibitor during in vitro activation of pHSCs induced significant downregulation of collagen type Ⅰ transcription. CONCLUSION: Our results emphasize the different regulation of miRNAs in in vitro and in vivo activated pHSCs. We also showed that miR-26a, 29a and 214 are involved in the regulation of collagen type I mRNA.
