A Fibrin Matrix Promotes the Differentiation of EMSCs Isolated from Nasal Respiratory Mucosa to Myelinating Phenotypical Schwann-Like Cells

Zhijian Zhang,Qian Chen,Jinbo Liu,Qinghua He,Yuepeng Zhou,Genbao Shao,Xianglan Sun,Xudong Cao,Aihua Gong,Ping Jiang 한국분자세포생물학회 2015 Molecules and cells Vol.38 No.3

Because Schwann cells perform the triple tasks of myelination, axon guidance and neurotrophin synthesis, they are candidates for cell transplantation that might cure some types of nervous-system degenerative diseases or injuries. However, Schwann cells are difficult to obtain. As another option, ectomesenchymal stem cells (EMSCs) can be easily harvested from the nasal respiratory mucosa. Whether fibrin, an important transplantation vehicle, can improve the differentiation of EMSCs into Schwann-like cells (SLCs) deserves further research. EMSCs were isolated from rat nasal respiratory mucosa and were purified using anti-CD133 magnetic cell sorting. The purified cells strongly expressed HNK-1, nestin, p75NTR, S-100, and vimentin. Using nuclear staining, the MTT assay and Western blotting analysis of the expression of cell-cycle markers, the proliferation rate of EMSCs on a fibrin matrix was found to be significantly higher than that of cells grown on a plastic surface but insignificantly lower than that of cells grown on fibronectin. Additionally, the EMSCs grown on the fibrin matrix expressed myelination-related molecules, including myelin basic protein (MBP), 2’,3’-cyclic nucleotide 3’-phosphodiesterase (CNPase) and galactocerebrosides (GalCer), more strongly than did those grown on fibronectin or a plastic surface. Furthermore, the EMSCs grown on the fibrin matrix synthesized more neurotrophins compared with those grown on fibronectin or a plastic surface. The expression level of integrin in EMSCs grown on fibrin was similar to that of cells grown on fibronectin but was higher than that of cells grown on a plastic surface. These results demonstrated that fibrin notonly promoted EMSC proliferation but also the differentiation of EMSCs into the SLCs. Our findings suggested that fibrin has great promise as a cell transplantation vehicle for the treatment of some types of nervous system diseases or injuries.

Genomic Sequence Analysis and Organization of BmKαTx11 and BmKαTx15 from Buthus martensii Karsch: Molecular Evolution of α-toxin genes

Xu, Xiuling,Cao, Zhijian,Sheng, Jiqun,Wu, Wenlan,Luo, Feng,Sha, Yonggang,Mao, Xin,Liu, Hui,Jiang, Dahe,Li, Wenxin Korean Society for Biochemistry and Molecular Biol 2005 Journal of biochemistry and molecular biology Vol.38 No.4

Based on the reported cDNA sequences of $BmK{\alpha}Txs$, the genes encoding toxin $BmK{\alpha}Tx11$ and $BmK{\alpha}Tx15$ were amplified by PCR from the Chinese scorpion Buthus martensii Karsch genomic DNA employing synthetic oligonucleotides. Sequences analysis of nucleotide showed that an intron about 500 bp length interrupts signal peptide coding regions of $BmK{\alpha}Tx11$ and $BmK{\alpha}Tx15$. Using cDNA sequence of $BmK{\alpha}Tx11$ as probe, southern hybridization of BmK genome total DNA was performed. The result indicates that $BmK{\alpha}Tx11$ is multicopy genes or belongs to multiple gene family with high homology genes. The similarity of $BmK{\alpha}$-toxin gene sequences and southern hybridization revealed the evolution trace of $BmK{\alpha}$-toxins: $BmK{\alpha}$-toxin genes evolve from a common progenitor, and the genes diversity is associated with a process of locus duplication and gene divergence.

Alternative Splicing of Breast Cancer Associated Gene BRCA1 from Breast Cancer Cell Line

Lixia, Miao,Zhijian, Cao,Chao, Shen,Chaojiang, Gu,Congyi, Zheng Korean Society for Biochemistry and Molecular Biol 2007 Journal of biochemistry and molecular biology Vol.40 No.1

Breast cancer is the most common malignancy among women, and mutations in the BRCA1 gene produce increased susceptibility to these malignancies in certain families. In this study, the forward 1-13 exons of breast cancer associated gene BRCA1 were cloned from breast cancer cell line ZR-75-30 by RT-PCR method. Sequence analysis showed that nine BRCA1 splice forms were isolated and characterized, compared with wild-type BRCA1 gene, five splice forms of which were novel. These splice isoforms were produced from the molecular mechanism of 5' and 3' alternative splicing. All these splice forms deleting exon 11b and the locations of alternative splicing were focused on two parts:one was exons 2 and 3, and the other was exons 9 and 10. These splice forms accorded with GT-AG rule. Most these BRCA1 splice variants still kept the original reading frame. Western blot analysis indicated that some BRCA1 splice variants were expressed in ZR-75-30 cell line at the protein level. In addition, we confirmed the presence of these new transcripts of BRCA1 gene in MDA-MB-435S, K562, Hela, HLA, HIC, H9, Jurkat and human fetus samples by RT-PCR analysis. These results suggested that breast cancer associated gene BRCA1 may have unexpectedly a large number of splice variants. We hypothesized that alternative splicing of BRCA1 possibly plays a major role in the tumorigenesis of breast and/or ovarian cancer. Thus, the identification of cancer-specific splice forms will provide a novel source for the discovery of diagnostic or prognostic biomarkers and tumor antigens suitable as targets for therapeutic intervention.

A Fibrin Matrix Promotes the Differentiation of EMSCs Isolated from Nasal Respiratory Mucosa to Myelinating Phenotypical Schwann-Like Cells

Chen, Qian,Zhang, Zhijian,Liu, Jinbo,He, Qinghua,Zhou, Yuepeng,Shao, Genbao,Sun, Xianglan,Cao, Xudong,Gong, Aihua,Jiang, Ping Korean Society for Molecular and Cellular Biology 2015 Molecules and cells Vol.38 No.3

Because Schwann cells perform the triple tasks of myelination, axon guidance and neurotrophin synthesis, they are candidates for cell transplantation that might cure some types of nervous-system degenerative diseases or injuries. However, Schwann cells are difficult to obtain. As another option, ectomesenchymal stem cells (EMSCs) can be easily harvested from the nasal respiratory mucosa. Whether fibrin, an important transplantation vehicle, can improve the differentiation of EMSCs into Schwann-like cells (SLCs) deserves further research. EMSCs were isolated from rat nasal respiratory mucosa and were purified using anti-CD133 magnetic cell sorting. The purified cells strongly expressed HNK-1, nestin, $p75^{NTR}$, S-100, and vimentin. Using nuclear staining, the MTT assay and Western blotting analysis of the expression of cell-cycle markers, the proliferation rate of EMSCs on a fibrin matrix was found to be significantly higher than that of cells grown on a plastic surface but insignificantly lower than that of cells grown on fibronectin. Additionally, the EMSCs grown on the fibrin matrix expressed myelination-related molecules, including myelin basic protein (MBP), 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNPase) and galactocerebrosides (GalCer), more strongly than did those grown on fibronectin or a plastic surface. Furthermore, the EMSCs grown on the fibrin matrix synthesized more neurotrophins compared with those grown on fibronectin or a plastic surface. The expression level of integrin in EMSCs grown on fibrin was similar to that of cells grown on fibronectin but was higher than that of cells grown on a plastic surface. These results demonstrated that fibrin not only promoted EMSC proliferation but also the differentiation of EMSCs into the SLCs. Our findings suggested that fibrin has great promise as a cell transplantation vehicle for the treatment of some types of nervous system diseases or injuries.

Genomic Sequence Analysis and Organization of BmKαTx11 and BmKαTx15from Buthus martensii Karsch: Molecular Evolution of α-toxin genes

XiuLing Xu,Feng Luo,WenXin Li,ZhiJian Cao,JiQun Sheng,WenLan Wu,YongGang Sha,Xin Mao,Hui Liu,DaHe Jiang 한국생화학분자생물학회 2005 BMB Reports Vol.38 No.4

Establishment and evaluation of a murine anb3-integrinexpressing cell line with increased susceptibility to Foot-and-mouth disease virus

Wei Zhang,Kaiqi Lian,Fan Yang,Yang Yang,Zhijian Zhu,Zixiang Zhu,Weijun Cao,Ruoqing Mao,Ye Jin,Jijun He,Jianhong Guo,Xiangtao Liu,Haixue Zheng 대한수의학회 2015 Journal of Veterinary Science Vol.16 No.3

Integrin anb3 plays a major role in various signaling pathways, cell apoptosis, and tumor angiogenesis. To examine the functions and rolesof anb3 integrin, a stable CHO-677 cell line expressing the murine anb3 heterodimer (designated as “CHO-677-manb3” cells) wasestablished using a highly efficient lentiviral-mediated gene transfer technique. Integrin subunits an and b3 were detected at the gene andprotein levels by polymerase chain reaction (PCR) and indirect immunofluorescent assay (IFA), respectively, in the CHO-677-manb3 cellline at the 20th passage, implying that these genes were successfully introduced into the CHO-677 cells and expressed stably. A plaque-formingassay, 50% tissue culture infective dose (TCID50), real-time quantitative reverse transcription-PCR, and IFA were used to detect the replicationlevels of Foot-and-mouth disease virus (FMDV) in the CHO-677-manb3 cell line. After infection with FMDV/O/ZK/93, the cell line showeda significant increase in viral RNA and protein compared with CHO-677 cells. These findings suggest that we successfully established a stableanb3-receptor-expressing cell line with increased susceptibility to FMDV. This cell line will be very useful for further investigation of anb3integrin, and as a cell model for FMDV research.