摘要：
To investigate the potential role of stromal cell‐derived factor‐1α (SDF‐1α)/CXCR4/ CXCR7 axis in adipose‐derived mesenchymal stem cells (ADSCs), quantitative reverse transcription‐polymerase chain reaction (qRT‐PCR) was employed to screen the effective small interfering RNA against CXCR4 and CXCR7 in ADSCs. The messenger RNA (mRNA) and proteins abundances of AKT (p‐AKT), ERK (p‐ERK), JNK (p‐JNK), and p38 (p‐p38) in different groups were identified by qRT‐PCR, western blot, and immunofluorescence staining method. Meanwhile, cell migration and cell proliferation with SDF‐1 treated were examined by a 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide assay and transwell permeable assay, respectively. Moreover, the interaction between CXCR4 and CXCR7 was examined by a GST pull‐down assay. CXCR4 small interfering RNA3 (siRNA3) and CXCR7 siRNA3 have been proved to the most effective tools for knockdown CXCR4 and CXCR7 expressions. mRNA abundance of JNK and p38 could be affected by SDF‐1α/CXCR4/CXCR7 axis. However, western blot analysis of p‐AKT, p‐ERK, p‐JNK, and p‐p38 in CXCR43‐treated ADSCs was significantly higher than that in the control group. Moreover, the immunofluorescence staining analysis revealed that the expressions of p‐ATK and p‐JNK proteins were significantly higher in NC‐ and SDF‐1‐treated subgroups than that in the CXCR4 and CXCR7 groups. p‐ATK and p‐JNK proteins in CXCR4 group were similar to that in CXCR7 group. Cell migration analysis of CXCR4‐treated ADSCs suggested that knockdown CXCR4 could effectively promote cell migration (p < .05). Moreover, CXCR4 could interact with CXCR7. The results in this study could provide a better understanding of SDF‐1α/CXCR4/CXCR7 axis during ADSCs development.
KEYWORDS：ADSCs, qRT-PCR, SDF-1α/CXCR4/CXCR7 axis, western blot
合作部分结果：
伯信合作技术：
qRT-PCR；GST Pulldown； Western Blot
原文链接：
