Induction of anoikis following myoblast transplantation into SCID mouse muscles requires the Bit1 and FADD pathways
Induction of anoikis following myoblast transplantation into SCID mouse muscles requires the Bit1 and FADD pathways
复制标题
DOI：
10.1111/j.1600-6143.2007.01830.x
复制
发表时间：
2007-06-01
影响因子：
8.8
通讯作者：
Tremblay, J. P.
中科院分区：
文献类型：
--
作者：
Bouchentouf, M.;Benabdallah, B. F.;Tremblay, J. P.
移植到小鼠肌肉中的75%的成肌细胞在移植后的前4天内死亡。本研究的目的是确定失巢凋亡是否在这种现象中发挥作用。Hoescht-PI标记法检测成肌细胞的存活率，细胞计数法检测成肌细胞的增殖能力。通过在SCID小鼠肌肉中注射用C-14-胸苷标记的人雄性成肌细胞来定量体内细胞存活和增殖。通过荧光显微镜和人Y染色体DNA定量PCR检测移植成肌细胞的存活和增殖。包含细胞外基质蛋白纤连蛋白提高了1.7倍，而玻连蛋白提高了1.8倍，移植成肌细胞的生存。FADD和Bit 1表达的降低降低了体外失巢凋亡，并改善了体内注射的成肌细胞存活。抗凋亡蛋白Bcl-2的异位表达在体外完全消除了成肌细胞失巢凋亡，并在体内将细胞存活率提高了3.1倍。在我看来，移植后的细胞死亡部分由失巢凋亡介导。包含细胞外基质蛋白增强了存活和增殖。减少促凋亡蛋白Bit 1和FADD的表达或过度表达Bcl-2改善成肌细胞存活。
Seventy-five percent of the myoblasts transplanted in the mouse muscle die during the first 4 days following transplantation. The purpose of this study was to determine if anoikis plays a role in this phenomenon. Survival and proliferation of myoblasts in vitro were determined by Hoescht-PI labeling and cell counts respectively. In vivo cell survival and proliferation were quantified by injecting human male myoblasts labeled with C-14-thymidine in SCID mouse muscles. Survival and proliferation of the transplanted myoblasts were evaluated by scintigraphy and quantitative PCR of human Y chromosomal DNA. Inclusion of the extracellular matrix protein fibronectin enhanced transplanted myoblast survival by 1.7-fold while vitronectin improved their proliferation by 1.8-fold. Reductions in FADD and Bit1 expression reduced anoikis in vitro and improved the injected myoblast survival in vivo. Ectopic expression of the anti-apoptotic protein Bcl-2 completely abolished myoblast anoikis in vitro and enhanced cell survival by 3.1-fold in vivo. Cell death following transplantation appears to me mediated in part by anoikis. Inclusion of extracellular matrix proteins enhanced both survival and proliferation. Reduced expression of the proapoptotic proteins Bit1 and FADD or overexpression of Bcl-2 improved myoblast survival.