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目的探讨在体外模拟的肿瘤微环境中,骨髓间充质干细胞(Mesenchymal stem cells,MSCs)中STAT3的过度表达和激活对其恶性转变的影响。方法通过MSCs分别与C6胶质瘤及星型胶质细胞间接共培养,模拟MSCs生长的肿瘤微环境,实验设实验组、阳性和阴性及空白对照组,MTT法检测各组细胞的增殖情况;RT-QPCR检测各组细胞中STAT3、CyclinD1及BCL-xl基因mRNA的表达水平;Western blot检测各组细胞中STAT3、P-STAT3、CyclinD及BCL-xl的蛋白表达水平;病理HE染色检测各组细胞注入裸鼠皮下成瘤情况。结果实验组MSCs生长接触抑制明显减弱,增殖活性增高;实验组STAT3、CyclinD1及BCL-xl基因mRNA的表达水平均明显高于阴性对照组和空白对照组,差异有统计学意义(P<0.05),与阳性对照组比较,差异无统计学意义(P>0.05);实验组STAT3、P-STAT3、CyclinD1及BCL-xl的蛋白表达水平明显高于阴性对照组和空白对照组,差异有统计学意义(P<0.05),与阳性对照组比较,差异无统计学意义(P>0.05);HE染色结果显示,实验组与阳性对照组细胞深染,聚集明显,排列不规则,细胞核大深染,瘤内有大量新生血管增生,可见组织坏死。结论 STAT3及P-STAT3的过度表达和激活,可能是造成MSCs在肿瘤微环中恶性转变的重要原因之一。
Objective To investigate the effect of overexpression and activation of STAT3 on the malignant transformation of mesenchymal stem cells (MSCs) in a tumor microenvironment simulated in vitro. Methods MSCs were co-cultured with C6 glioma and astrocyte respectively to simulate the growth of MSCs. The experimental groups were divided into experimental group, positive and negative groups and blank control group. MTT assay was used to detect the proliferation of each group. The expression of STAT3, CyclinD1 and BCL-xl mRNA in each group was detected by RT-QPCR. The protein expression of STAT3, P-STAT3, CyclinD and BCL-xl in each group was detected by Western blot. Cells into nude mice subcutaneous tumor formation. Results The contact inhibition of MSCs in experimental group was significantly decreased and the proliferation activity was increased. The mRNA expression levels of STAT3, CyclinD1 and BCL-xl in experimental group were significantly higher than those in negative control group and blank control group (P <0.05) (P> 0.05). The protein expression levels of STAT3, P-STAT3, CyclinD1 and BCL-xl in the experimental group were significantly higher than those in the negative control group and the blank control group, the difference was statistically significant (P <0.05). Compared with the positive control group, the difference was not statistically significant (P> 0.05). The results of HE staining showed that the cells in the experimental group and the positive control group were deeply stained, clustered obviously, arranged irregularly, , A large number of tumor neovascularization, visible tissue necrosis. Conclusions The overexpression and activation of STAT3 and P-STAT3 may be one of the important reasons for the malignant transformation of MSCs in the microrings of tumor.