太白楤木皂苷下调sirtuin 1表达对HEC-1B细胞侵袭和迁移的影响

Saponins of Aralia Taibaiensis inhibits invasion and migration of HEC-1B cell line through down-regulating the expression of Sirtuin 1

目的太白楤木皂苷(SAT)具有明显的抗肿瘤功能,本研究通过探究SAT对子宫内膜癌细胞HEC-1B增殖、迁移、侵袭的影响,以期为子宫内膜癌的治疗提供理论依据。方法实验分为对照组(未加药物)、阳性对照组(30μmol/L环磷酰胺)、SAT 10μmol/L组、SAT 20μmol/L组、SAT 30μmol/L组。以不同浓度SAT及环磷酰胺处理HEC-1B细胞后,采用CCK8法检测HEC-1B细胞增殖情况,平板细胞克隆形成实验检测菌落形成;Transwell小室试验检测SAT对HEC-1B细胞迁移、侵袭能力的影响,实时荧光定量(qRT-PCR)法检测去沉默调控蛋白-1(sirtuin 1)mRNA相对表达量,蛋白免疫印迹法(Western blotting)检测sirtuin 1蛋白、金属基质蛋白酶-2(MMP-2)、金属基质蛋白酶-9(MMP-9)表达。结果与对照组相比,阳性对照组、SAT处理组细胞增殖、侵袭、迁移数量显著降低(均P<0.05),且呈剂量依赖;与阳性对照组相比,SAT 20μmol/L组、30μmol/L组细胞增殖、侵袭、迁移数量显著降低(均P<0.05);与对照组相比,阳性对照组、SAT处理组MMP-2、MMP-9、sirtuin 1 mRNA、sirtuin 1蛋白水平显著降低(均P<0.05),且随SAT浓度升高而下降;与阳性对照组相比,SAT 20μmol/L组、SAT 30μmol/L组MMP-2、MMP-9、sirtuin 1 mRNA、sirtuin 1蛋白水平显著降低(均P<0.05)。结论太白楤木皂苷可抑制子宫内膜癌细胞HEC-1B细胞迁移、侵袭,可能与下调sirtuin 1表达有关,能为子宫内膜癌的治疗提供新可能。

Objective To explore the effects of Saponins of Aralia Taibaiensis(SAT) on the proliferation, migration and invasion of endometrial cancer cell HEC-1 B, and to provide a theoretical basis for the treatment of endometrial cancer. Methods HEC-1 B cells were treated with different concentrations SAT(0, 10, 20, 30 μmol/L). CCK8 assay was used to detect the cell proliferation, plate cell clone formation assay was used to detect colony formation, transwell chamber test was used to detect migration and invasion, and real-time PCR was used to detect the expression of sirtuin-1. The protein expressions of sirtuin 1, MMP-2 and MMP-9 were detected by Western Blotting. Results Compared with the control group, the cell proliferation, invasion and migration in SAT treatment group and the positive control group decreased significantly(all P<0.05), which was dose-dependent. Compared with the positive control group, the cell proliferation, invasion and migration in 20 and 30 μmol/L SAT group decreased significantly(all P<0.05). Compared with the control group, the levels of sirtuin 1 mRNA and protein, MMP-2 and MMP-9 protein in SAT treatment group and the positive control group decreased significantly, which was dose-dependent. Compared with the positive control group, the levels of sirtuin 1 mRNA and protein, MMP-2 and MMP-9 protein in 20 and 30 μmol/L SAT group decreased significantly(all P<0.05). Conclusion SAT can inhibit the migration and invasion of HEC-1 B cells, which may be related to the down-regulation of sirtuin-1 expression, and may provide a new possibility for the treatment of endometrial cancer.