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目的:探讨人骨髓间充质干细胞(human bone marrow-derived mesenchymal stem cell,hBMSC)对胃癌细胞的趋化性,为将MSC研发成为胃癌基因治疗载体提供实验依据。方法:骨髓培养法分离培养bBMSC并进行流式细胞术鉴定。Transwell实验检测hBMSC对胃癌MKN45细胞和人成纤维细胞(humanfibroblast,hFB)的趋化能力。建立MKN45细胞裸鼠移植瘤模型,感染Lenti-EGFP(MOI为50)的hBMSC或hFB细胞经尾静脉注射入荷瘤小鼠,荧光显微镜下观察移植瘤组织及各脏器GFP的表达。结果:培养第3代的hBMSC细胞CD44、CD105阳性率为(96.7±1.84)%、(98.1±0.95)%,而CD34、CD45表达阴性。hBMSC细胞向MKN45细胞的趋化能力明显强于胃上皮细胞GES-1组及空白对照细胞[(239.5±54.3)vs(43.57±4.6)、(37.3±4.7)个,P<0.01],且hBMSC向MKN45细胞的趋化能力明显强于hFB细胞向MKN45细胞的趋化能力[(239.5±54.3)vs(27.7±16.7),P<0.01]。与hFB相比,hBMSC对胃癌移植瘤组织具有明显趋向性;hBMSC组移植瘤组织内可见GFP表达,移植瘤小鼠部分肝脏(20%)及肺脏(20%)有GFP表达,但较移植瘤组织内GFP表达率和强度均低(P<0.05)。结论:hBMSC在体内外对胃癌细胞均有特异性趋化作用,有望研发为胃癌基因治疗的良好载体。
OBJECTIVE: To investigate the chemotaxis of human bone marrow-derived mesenchymal stem cells (hBMSC) to gastric cancer cells and to provide an experimental basis for developing MSC as a gene therapy vector for gastric cancer. Methods: BMSCs were isolated and cultured by bone marrow culture and identified by flow cytometry. The chemotaxis ability of hBMSC to gastric cancer MKN45 cells and human fibroblasts (hFB) was detected by Transwell assay. The nude mice bearing nude mice model of MKN45 cells were established. The hBMSC or hFB cells infected with Lenti-EGFP (MOI 50) were injected into the tumor-bearing mice through the tail vein and the expression of GFP in the transplanted tumor tissues and various organs was observed under a fluorescence microscope. Results: The positive rate of CD44 and CD105 in cultured hBMSC cells was (96.7 ± 1.84)%, (98.1 ± 0.95)%, while the expression of CD34 and CD45 was negative. The chemotactic capacity of hBMSC cells to MKN45 cells was significantly higher than that of GES-1 cells in gastric epithelial cells and blank control cells [(239.5 ± 54.3) vs (43.57 ± 4.6), (37.3 ± 4.7), P <0.01] Chemotaxis to MKN45 cells was significantly stronger than that of hFB cells to MKN45 cells [(239.5 ± 54.3 vs 27.7 ± 16.7, P <0.01]. Compared with hFB, hBMSC had obvious tendency to the transplanted tumor of gastric cancer. The expression of GFP was found in the transplanted tumor of hBMSC group. The expression of GFP was observed in some liver (20%) and lung (20% Tissue GFP expression rate and intensity were low (P <0.05). Conclusion: hBMSC has specific chemotactic effect on gastric cancer cells both in vitro and in vivo, which is expected to be a good carrier for gene therapy of gastric cancer.