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目的探讨多氯联苯(PCB126)对大鼠骨髓间充质干细胞(MSCs)c-fos和c-jun表达的影响。方法采用percoll分离液(密度1.073g/cm3)分离Wistar大鼠MSCs、培养并传代,实验分为3组:对照组、低剂量组(10-7mol/L PCB126)和高剂量组(10-6mol/L PCB126)。PCB126分别作用于MSCs 12h和24h后,利用MTT方法检测细胞的增殖率,免疫荧光化学方法检测细胞c-fos的表达;PCB126分别作用于MSCs 30min、1h、2h、4h、8h、12h和24h后,利用RT-PCR分析c-fos和c-jun mRNA的表达;Western blot方法分析细胞c-fos和c-jun蛋白的表达。结果 10-6mol/L和10-7mol/L组12h时细胞的增殖率分别为13.8%和19.1%,24h时分别为31.5%和36.1%,48h时分别为42.5%和43.6%(t=8.42,P<0.01)。10-6mol/L和10-7mol/L组12h时c-fos的阳性率分别为54.6%(231/446)和51.3%(214/416)(χ2=15.59,P<0.01;χ2=15.45,P<0.01),24h时c-fos的阳性率分别为83.2%(376/425)和73.0%(309/423)(χ2=60.56,P<0.01;χ2=25.79,P<0.01)。RT-PCR结果显示,10-6mol/L组和10-7mol/L组在30 min和1h时c-fos mRNA的表达显著上调;两组在不同时间c-jun mRNA均上调且10-6mol/L组的表达水平明显高于10-7mol/L组(t=7.88,P<0.01)。10-6mol/L和10-7mol/L组12h时c-fos和c-jun蛋白表达上调(t=9.91和t=8.84,P<0.01),24h时c-fos和c-jun表达显著上调(t=9.52,P<0.01)。结论 PCB126能促进MSCs增殖,上调癌相关基因c-fos和c-jun的表达,PCB126影响MSCs的功能与c-fos和c-jun的表达异常有关。
Objective To investigate the effects of PCB126 on the expression of c-fos and c-jun in rat bone marrow mesenchymal stem cells (MSCs). Methods Wistar rat MSCs were isolated and cultured with percoll (density: 1.073 g / cm3), cultured and passaged. The experiment was divided into three groups: control group, low dose group (10-7mol / L PCB126) and high dose group / L PCB126). PCB126 cells were treated with PCB126 for 12h and 24h respectively, MTT assay was used to detect the proliferation rate of cells, and the expression of c-fos was detected by immunofluorescence staining. PCB126 cells were treated with MSCs for 30min, 1h, 2h, 4h, 8h, 12h and 24h respectively The expression of c-fos and c-jun mRNA was analyzed by RT-PCR. The expression of c-fos and c-jun protein was analyzed by Western blot. Results The proliferation rates were 13.8% and 19.1% at 12 h in 10-6 mol / L and 10-7 mol / L groups respectively, 31.5% and 36.1% at 24 h and 42.5% and 43.6% at 48 h (t = 8.42 , P <0.01). The positive rates of c-fos in 10-6mol / L and 10-7mol / L groups were 54.6% (231/446) and 51.3% (214/416) respectively (χ2 = 15.59, P <0.01; χ2 = 15.45, The positive rates of c-fos at 24h were 83.2% (376/425) and 73.0% (309/423) respectively (χ2 = 60.56, P <0.01; χ2 = 25.79, P <0.01). The results of RT-PCR showed that the expression of c-fos mRNA in 10-6mol / L group and 10-7mol / L group was significantly up-regulated at 30min and 1h; The c-jun mRNA was up-regulated in both groups at 10-6mol / L group was significantly higher than the 10-7mol / L group (t = 7.88, P <0.01). The expressions of c-fos and c-jun were up-regulated at 10h and 10-7mol / L for 12h (t = 9.91 and t = 8.84, P <0.01) (t = 9.52, P <0.01). Conclusions PCB126 can promote the proliferation of MSCs and up-regulate the expression of c-fos and c-jun. The effect of PCB126 on MSCs is related to the abnormal expression of c-fos and c-jun.