Msp对人非小细胞肺癌细胞增殖、迁移和侵袭能力影响的 实验研究

目的 探讨巨噬细胞刺激蛋白(macrophage stimulating protein, Msp)对人非小细胞肺癌细胞株PC14增殖、迁移和侵袭能力影响的研究。方法 构建Msp编码基因st1 真核表达载体,将st1 导入Msp（-）和RON（-）人非小细胞肺癌细胞株PC14,并用G418筛选出稳定表达的细胞株。通过RT-PCR方法检测已转染st1 载体PC14细胞中st1 的表达,用Western blot检测Msp在PC14、PC14-st1 -pEGFP-N1、PC14-pEGFP-N1中的表达水平,同时检测RON在PC14和RON(+)细胞SKBR-3中的表达情况。通过计算RAW 264.7（小鼠单核巨噬细胞）和SKBR-3细胞在各转染组细胞培养上清液的刺激下穿过Transwell 微孔膜的细胞数来评价Msp的生物学活性。利用MTT法检测Msp对PC14细胞增殖力的影响,采用Transwell小室和基质胶侵袭实验观察Msp对PC14细胞迁移和侵袭能力的影响。结果 转染st1 基因后,形成PC14-st1 -pEGFP-N1,稳定表达st1 mRNA及蛋白,其本身的增殖能力被明显抑制,迁移能力和侵袭能力均较PC14和PC14-pEGFP-N1降低。它还通过旁分泌的方式对RON (+)阳性的癌细胞RAW264.7和SKBR-3的迁移起促进作用。结论 Msp表达可降低人非小细胞肺癌细胞PC14的增殖、迁移和侵袭能力,却对RON (+)阳性的癌细胞迁移通过旁分泌的方式有促进作用。     

骨髓间充质干细胞向软骨及骨分化：Wnt5a/PCP信号通路作用的研究与进展

BACKGROUND: Wnt5a is able to inhibit canonical Wnt signaling and activate non-canonical Wnt signaling pathway. In recent years, it has been found that non-classical Wnt5a/PCP signaling pathway mediated by Wnt5a plays an important role in the process of bone marrow mesenchymal stem cell proliferation and differentiation, but the underlying mechanism is unclear. OBJECTIVE: To summarize the progress in downstream effector molecules related to Wnt5a/PCP signaling pathway, and its roles in the chondrogenic and osteogenic differentiation of bone marrow mesenchymal stem cells. METHODS: A computer-based online search of CqVip, CNKI and PubMed databases between January 2000 and February 2016 was performed using the Chinese keywords of “BMSCs, Wnt signaling pathways, chondrogenic differentiation, osteogenic differentiation” and English keywords of “BMSCs, chondrogenic differentiation, osteogenic differentiation, Wnt, Fzd, Ror2, RhoA, ROCK, JNK”, respectively. Literatures related to bone marrow mesenchymal stem cell chondrogenic and osteogenic differentiation were selected. Finally, 43 eligible articles were included for analysis through excluding the old and repeated research. RESULTS AND CONCLUSION:Wnt5a, a representative protein in non-canonical Wnt signaling pathway, paticipates in the cytoskeleton, cell migration and cell polarization and other activities by mediating its downstream signaling molecules such as Fzd, Ror, RhoA, ROCK, JNK, thereby regulating its proliferation and differentiation. But it is unclear how Wnt5a/PCP participates in the bone marrow mesenchymal stem cell chondrogenic and osteogenic differentiation and how the downstream effector molecules interact or function independently, which requires further studies. 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

MSP影响非小细胞肺癌PC14细胞周期和上皮间质转化的研究

目的研究巨噬细胞刺激蛋白（MSP）在无巨噬细胞刺激蛋白受体（RON）的情况下对非小细胞肺癌PC14细胞周期的影响,并分析其对PC14上皮间质转化（EMT）的影响。方法取MSP阴性、RON阴性的PC14细胞系,稳定表达MSP的细胞系PC14-Mst1-pEGFP-N1和稳定表达荧光蛋白的空载质粒细胞系PC14-pEGFP-N1,流式细胞术检测各细胞系细胞周期的影响;透射电镜观察在细胞增殖期间细胞之间的连接间隙;逆转录（RT）-PCR和Western blot分别检测E-钙粘蛋白(E-cadherin)和波形蛋白(Vimentin) mRNA、蛋白表达水平。结果流式细胞术结果显示与PC14、PC14-pEGFP-N1相比,PC14-Mst1-pEGFP-N1 细胞G1/G0期数量增多,S期与G2/M期减少;透射电镜发现PC14-Mst1-pEGFP-N1细胞之间的细胞连接较紧密,细胞之间空隙距离缩小;RT-PCR和Western blot检测结果显示,与PC14细胞相比,PC14-Mst1-pEGFP-N1细胞E-cadherin的mRNA和蛋白表达水平升高,而Vimentin的mRNA、蛋白表达水平降低。结论MSP在RON阳性的情况下,可使G1期肺癌细胞增多、分裂相减少,抑制EMT。     

力学刺激调控骨髓间充质细胞向软骨的分化

BACKGROUND:Bone marrow mesenchymal cells are ideal seed cells for tissue engineering and cell therapy. We are seeking to improve the cartilage repair strategy based on bone marrow mesenchymal cells in vitro by use of natural mechanical transfer pathways. OBJECTIVE:To analyze the mechanical factors affecting the fate of bone marrow mesenchymal cells and utilize the mechanical strategy to induce chondrogenesis. METHODS:The first author retrieved the literature in CNKI, PubMed, Medline and Elsevier databases (1985-2015) with the key words of “mechanical regulation, bone marrow mesenchymal cells, bone marrow mesenchymal stem cells, chondrogenic differentiation, cartilage differentiation”. RESULTS AND CONCLUSION:Herein, we summarize some latest research results on the mechanical stimulus-induced chondrogenic differentiation of bone marrow mesenchymal cells, discuss the effects of different mechanical stimulations on the chondrogenic differentiation of bone marrow mesenchymal cells, as well as summarize the biological mechanism and signal pathways of the cells in response to mechanical stimulations in the process of chondrogenic differentiation. At last, this paper points out the future prospects of the key problems in this field, such as cartilage regeneration and repair strategy based on chondrogenic differentiation of bone marrow mesenchymal cells under mechanical stimulation.