CD151对人脐带来源间充质干细胞生物学特性的影响

 目的：探讨CD151对人脐带来源间充质干细胞（human umbilical cord mesenchymal stem cells，hUC-MSCs）生物学性状的影响。方法：使用siRNA干扰hUC-MSCs 表面CD151，实验分为实验组（siRNA-CD151组）和对照组（siRNA-NC组）；流式细胞术检测干扰72 h后hUC-MSCs CD151及其它细胞表型,von Kossa和油红O染色检测体外成骨、成脂诱导分化情况；流式细胞术分析细胞周期变化；real-time PCR检测hUC-MSCs CD151、肝细胞生长因子（hepatocyte growth factor，HGF）、转化生长因子β1（transforming growth factor β1，TGF-β1）、环氧合酶2（cyclooxygenase 2，COX-2)和吲哚胺2，3-双加氧酶（indoleamine 2,3-dioxygenase，IDO）mRNA的表达量；ELISA检测hUC-MSCs HGF分泌量。结果：(1) 流式细胞术检测发现干扰72 h hUC-MSCs实验组CD151（11.97±2.63 vs 95.66±1.56；P<0.01）和CD105（93.66±0.21 vs 83.37±0.71；P<0.05）较对照组显著降低；real-time PCR 检测hUC-MSCs CD151 mRNA变化和流式结果一致。(2)给予siRNA-CD151后能够减缓细胞周期进展，表现为细胞G1期增多，S期减少。(3)实验组hUC-MSCs HGF和TGF-β1表达量低于对照组（P<0.01），而COX-2表达量升高（P<0.05），IDO无明显改变；ELISA检测显示实验组hUC-MSCs HGF分泌量较对照组下降（P<0.01）。结论：体外干扰CD151后hUC-MSCs 仍保持干细胞表型,体外成脂诱导分化能力无明显变化，但成骨诱导分化能力、增殖能力和相关免疫调节因子表达发生改变。

Effect of CD151 on biological characteristics of human umbilical cord mesenchymal stem cells

AIM：To study the effect of CD151 on the biological characteristics of human umbilical cord mesenchymal stem cells (hUC-MSCs). METHODS：CD151 expression on hUC-MSCs was interfered by siRNA. The cells were divided into siRNA-CD151 group and negative control group (treated with siRNA-NC). The efficiency of interference after 72 h and the changes of other surface markers were detected by flow cytometry. The ability of differentiation was assessed by oil red O and von Kossa staining. The cell cycle was analyzed by flow cytometry. The mRNA expression of CD151, hepatocyte growth factor (HGF), transforming growth factor β1 (TGF-β1), cyclooxygenase 2 (COX-2) and indoleamine 2, 3-dioxygenase (IDO) in hUC-MSCs was detected by real-time PCR. The secretion of HGF by hUC-MSCs was measured by ELISA. RESULTS：The results of flow cytometry showed that the expression of CD151 (11.97±2.63 vs 95.66±1.56, P<0.01) and CD105 (93.66±0.21 vs 83.37±0.71, P<0.05) on hUC-MSCs in siRNA-CD151 group was lower than that in negative control group. The consistent results were also achieved by using the method of real-time PCR. Treatment with siRNA-CD151 down-regulated the progress of the cell cycle as the G1 phase increased and the S phase decreased. The mRNA expression levels of HGF and TGF-β1 in hUC-MSCs in siRNA-CD151 group were lower than those in negative control group, and opposite result of COX-2 mRNA expression was observed. The IDO mRNA in hUC-MSCs was unchanged with IFN-γ stimulation for 24 h. HGF concentration in siRNA-CD151 group  was decreased as compared with negative control group. CONCLUSION：Interfering CD151 expression on hUC-MSCs doesn’t change other surface markers except CD105, and maintains the capacity of adipogenic differentiation. However, it changes the osteogenic differentiation, proliferation and the expression of immunomodulatory cytokines.