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目的:体外研究人Burkitt淋巴瘤细胞系Raji细胞和人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,hUC-MSC)间的相互作用,探讨MSC在恶性肿瘤治疗中的潜在应用前景。方法:将Raji细胞和hUC-MSC进行直接接触和非接触共培养,观察MSC对Raji细胞的作用,采用CCK-8法观察Raji细胞的增殖情况,流式细胞术检测Raji细胞周期的变化,同时应用siRNA干扰技术分析血管内皮生长因子(VEGF)在MSC向Raji细胞迁移过程中的作用(即用siRNA阻断Raji细胞VEGF的表达),ELISA法检测Raji细胞培养上清VEGF的分泌量,qRT-PCR法检测Raji细胞VEGF mRNA表达的变化。结果:MSC及其条件培养基(MSC-conditioned medi-um,MSC-CM)均促进Raji细胞的增殖,72 h MSC-CM培养的Raji细胞增殖活性为0.99±0.05,而对照组为0.71±0.07(P<0.01)。无论是直接接触还是条件培养基均促使Raji细胞从G0/G1期向S期的转化,MSC-CM的作用尤其明显,S期的细胞数目由16.33±1.37增加到28.50±1.41,而G0/G1期细胞数目则由77.70±1.57下降到54.40±1.57(P<0.01)。siRNA干扰Raji细胞后,VEGF无论是在蛋白水平还是mRNA水平都明显下降,且MSC向Raji细胞的趋化能力也显著降低(96.00±5.28 vs 143.00±7.20,P<0.01)。结论:MSC具有向Raji细胞趋化的作用,其机制与Raji细胞分泌VEGF呈正相关;MSC可促进Raji细胞从G0/G1期向S期转化,从而促进Raji细胞的增殖。
OBJECTIVE: To study the interaction between human Burkitt lymphoma cell line Raji cells and human umbilical cord mesenchymal stem cells (hUC-MSC) in vitro and to explore the potential application of MSCs in the treatment of malignant tumors. Methods: The Raji cells and hUC-MSC were directly contacted with non-contact co-culture to observe the effect of MSC on Raji cells. The proliferation of Raji cells was observed by CCK-8 method. The cell cycle of Raji cells was detected by flow cytometry. The effect of vascular endothelial growth factor (VEGF) on the migration of MSCs to Raji cells was analyzed by siRNA interference (ie, the expression of VEGF in Raji cells was blocked by siRNA). The secretion of VEGF in Raji cell culture supernatant was detected by ELISA. The expression of qRT- PCR assay Raji cell VEGF mRNA expression changes. Results: The proliferation of Raji cells was induced by MSC and its conditioned media (MSC-CM-CM). The proliferation of Raji cells incubated with MSC-CM for 72 h was 0.99 ± 0.05, while the control group was 0.71 ± 0.07 (P <0.01). The effect of MSC-CM was especially evident in either Raji cells from G0 / G1 phase to S phase, both in direct contact and conditioned media. The number of cells in S phase increased from 16.33 ± 1.37 to 28.50 ± 1.41, while G0 / G1 The number of stage cells decreased from 77.70 ± 1.57 to 54.40 ± 1.57 (P <0.01). After siRNA interference with Raji cells, VEGF significantly decreased both in protein and mRNA levels, and the chemotactic ability of MSC to Raji cells was also significantly decreased (96.00 ± 5.28 vs 143.00 ± 7.20, P <0.01). CONCLUSION: MSCs can chemotactic to Raji cells. The mechanism is related to the secretion of VEGF by Raji cells. MSCs can promote the transformation of Raji cells from G0 / G1 phase to S phase and promote the proliferation of Raji cells.