The effect of C3 transferase on human adipose-derived stem cells
The effect of C3 transferase on human adipose-derived stem cells
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DOI：
10.1007/s13577-011-0033-0
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发表时间：
2011-01-01
期刊：
影响因子：
4.3
通讯作者：
Kusumoto, Kenji
中科院分区：
文献类型：
--
作者：
Kakudo, Natsuko;Kushida, Satoshi;Kusumoto, Kenji
人脂肪源性干细胞（ASCs）是成体多能干细胞，具有分化为脂肪、软骨、骨或神经的能力，可应用于组织工程。另一方面，外泌酶C3转移酶（C3）是一种Rho抑制剂。一旦进入细胞质，穿透细胞的部分被释放，从而使C3转移酶在细胞内自由扩散，使RhoA、RhoB和RhoC失活，但不使相关的gtp酶如Cdc42或Rac1失活。在本研究中，我们采用免疫荧光染色法研究了添加C3诱导的ASC细胞骨架变化，采用实时RT-PCR法研究了α -平滑肌肌动蛋白（a-SMA）基因表达的变化，并采用下拉法研究了rho抑制作用。添加浓度为1 μ g/ml的C3显著降低应力纤维断裂和a- sma表达24 h，并降低Rho活性水平。虽然可以假设发生的相关性，但需要进一步的检查来验证它。在涉及Rho的ASC细胞骨架控制中，C3可能是细胞内信号传递的有效抑制剂。
Human adipose-derived stem cells (ASCs) are adult pluripotent stem cells, which have the ability to differentiate into fat, cartilage, bone, or nerves that can be applied in tissue engineering. On the other hand, the exoenzyme C3 transferase (C3) is a Rho inhibitor. Once in the cytosol, the cell-penetrating moiety is released, thereby allowing C3 transferase to freely diffuse intracellularly and inactivate RhoA, RhoB, and RhoC, but not related GTPases such as Cdc42 or Rac1. In this study, we investigated ASC cytoskeletal changes induced by the addition of C3 employing immunofluorescence staining, changes in alpha-smooth muscle actin (a-SMA) gene expression employing real-time RT-PCR, and the Rho-inhibitory effect employing the pull-down assay. C3 significantly reduced stress fiber disruption and a-SMA expression 24 h after its addition at a concentration of 1 mu g/ml, and it also reduced the Rho activity level. While the correlation of the occurrence can be assumed, it requires further examination to verify it. C3 may be an effective inhibitor of intracellular signal transmission in ASC cytoskeletal control involving Rho.