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目的初步探讨多聚(腺苷二磷酸核糖)水解酶[poly-(ADP-ribose)glycohydrolase,PARG]抑制对小鼠结肠癌CT26细胞VEGF-C蛋白表达影响。方法分为实验组(GLTN处理)与对照组(GLTN未处理),采用Western blot法检测2组小鼠结肠癌CT26细胞的PARG、PARP-1、NF-κB及VEGF-C的蛋白表达变化;RT-PCR分析实验组与对照组的VEGF-C mRNA水平变化;ELISA法检测2组小鼠结肠癌CT26细胞上清液VEGF-C蛋白表达变化。结果 PARG抑制后小鼠结肠癌CT26细胞表达的PARG和NF-κB蛋白较对照组下降(P<0.01),PARP-1和VEGF-C蛋白较对照组降低(P<0.01)。RT-PCR结果显示,实验组CT26细胞株VEGF-C mRNA表达[相对灰度值为(0.39±0.08)]较对照组[相对灰度值为(0.63±0.04)]显著下降(P<0.01)。实验组小鼠结肠癌CT26细胞分泌的VEGF-C蛋白量[(95.86±5.43)pg/ml]较对照组[(1 99.32±0.75)pg/ml]明显降低(P<0.05)。结论在小鼠结肠癌CT26细胞中,抑制PARG可下调PARP-1和NF-κB的表达,从而使VEGF-C表达下降,这可能在抑制肿瘤淋巴管形成中起重要作用。
Objective To investigate the effect of poly (ADP-ribose) glycohydrolase (PARG) on the expression of VEGF-C protein in mouse colon cancer CT26 cells. Methods The experimental group (GLTN treatment) and the control group (untreated GLTN) were used to detect the protein expressions of PARG, PARP-1, NF-κB and VEGF-C in two groups of colon cancer CT26 cells by Western blot. The changes of VEGF-C mRNA expression in experimental group and control group were detected by RT-PCR. The expression of VEGF-C protein in colon cancer CT26 cells was detected by ELISA. Results The expressions of PARG and NF-κB in CT26 cells of mice with PARG inhibition decreased compared with the control group (P <0.01). The protein expressions of PARP-1 and VEGF-C in the CT26 cells decreased significantly (P <0.01). The results of RT-PCR showed that the expression of VEGF-C mRNA in CT26 cell line was significantly lower than that in control group [(0.63 ± 0.04) vs (0.39 ± 0.08), P <0.01) . The amount of VEGF-C protein secreted by colon cancer CT26 cells in experimental group [(95.86 ± 5.43) pg / ml] was significantly lower than that in control group [(99.32 ± 0.75) pg / ml] Conclusion Inhibition of PARG can down-regulate the expression of PARP-1 and NF-κB in mouse colon cancer CT26 cells, thereby decreasing the expression of VEGF-C, which may play an important role in the inhibition of lymphangiogenesis.