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目的 :探讨人脐带间充质干细胞hUCMSCs(human umbilical cord mesenchymal stem cells)在U251脑胶质瘤微环境中是否存在STAT3(signal transducer and activator of transcription 3)过度激活及细胞生物学行为改变。方法:本实验将不同处理的细胞分为3组:空白对照组(h UCMSCs单独培养)、实验组(hUCMSCs与U251共培养)、阳性对照组(U251单独培养)。流式细胞术检测细胞周期;RT-qPCR检测STAT3及其下游原癌基因c-Myc和抗凋亡基因Bcl-xl的m RNA的表达;Western blot及免疫荧光检测激活状态的p-STAT3及c-Myc和Bcl-xl蛋白表达及定位;Transwell实验检测细胞迁移和侵袭能力。结果:实验组h UCMSCs排列紊乱,成簇分布,呈现轻度异型性;实验组细胞周期G1期比例显著低于对照组(P=0.000);实验组STAT3、c-Myc和Bcl-xl的mRNA表达水平均明显高于空白对照组(P<0.05);实验组p-STAT3、STAT3、c-Myc和Bcl-xl的蛋白表达明显高于空白对照组(P<0.05),且主要定位于细胞核内。实验组细胞的迁移和侵袭能力均显著高于对照组(P<0.05)。结论:hUCMSCs在U251胶质瘤微环境中存在STAT3的过度激活及生物学行为改变,出现恶性转变倾向。
OBJECTIVE: To investigate the signal transducer and activator of transcription 3 (STAT3) overexpression and the biological behavior of human umbilical cord mesenchymal stem cells (UU1) in U251 glioma microenvironment. Methods: The cells in different treatment groups were divided into three groups: blank control group (h UCMSCs cultured alone), experimental group (hUCMSCs co-cultured with U251) and positive control group (U251 cultured alone). Flow cytometry was used to detect the cell cycle. RT-qPCR was used to detect the expression of STAT3 and its downstream oncogene c-Myc and anti-apoptotic gene Bcl-xl m RNA. Western blot and immunofluorescence were used to detect the activation of p-STAT3 and c -Myc and Bcl-xl protein expression and localization; Transwell assay to detect cell migration and invasion ability. Results: The expression of STAT3, c-Myc and Bcl-xl mRNA in the experimental group was significantly different from that in the control group (P = 0.000). The percentage of G1 phase in the experimental group was significantly lower than that in the control group (P <0.05). The protein expression of p-STAT3, STAT3, c-Myc and Bcl-xl in the experimental group was significantly higher than that in the blank control group (P <0.05), and mainly localized in the nucleus Inside. Migration and invasion ability of the experimental group were significantly higher than the control group (P <0.05). CONCLUSION: The overexpression of STAT3 and biological behavior in hUCMSCs in U251 glioma microenvironment have the tendency of malignant transformation.