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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 factorβ1,TGF-β1)、环氧合酶2(cy-clooxygenase 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 vs95.66±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。
Objective: To investigate the effect of CD151 on biological characteristics of human umbilical cord-derived mesenchymal stem cells (hUC-MSCs). METHODS: The CD151 on the surface of hUC-MSCs was induced by siRNA. The experiment was divided into experimental group (siRNA-CD151 group) and control group (siRNA-NC group). Flow cytometry was used to detect CD151 and other cell phenotypes , von Kossa and oil red O were used to detect the osteogenic and adipogenic differentiation in vitro. The cell cycle was analyzed by flow cytometry. The expressions of CD151, hepatocyte growth factor (HGF) Transforming growth factor β1 (TGF-β1), cyclooxygenase 2 (COX-2) and indoleamine 2,3-dioxygenase (IDO ) mRNA expression; ELISA detection of hUC-MSCs HGF secretion. Results: (1) Flow cytometry showed that 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) The control group was significantly reduced; real-time PCR detection hUC-MSCs CD151 mRNA changes consistent with the flow results. (2) The siRNA-CD151 could slow the cell cycle progression, showing the increase of G1 phase and the decrease of S phase. (3) The expression of HGF and TGF-β1 in hUC-MSCs in experimental group was lower than that in control group (P <0.01), while the expression of COX-2 increased (P <0.05) -MSCs HGF secretion than the control group decreased (P <0.01). CONCLUSION: The hUC-MSCs still maintain the phenotype of stem cells after in vitro interference with CD151, and have no obvious change in adipogenic differentiation in vitro. However, the osteogenic differentiation, proliferation and the expression of related immunoregulatory factors are changed.