黄芪多糖对三阴性乳腺癌细胞增殖和侵袭作用的研究

Effect of Astragalus Polysaccharide on proliferation and invasion of triple negative breast cancer cells

目的 探索三阴性乳腺癌细胞应用黄芪多糖(APS)处理后,细胞增殖和侵袭能力的变化,及其对Wnt/β-catenin通路的影响。方法 将BT-549细胞分为BT-549空白组(常规培养)和BT-549低、中、高剂量实验组(分别给予200、400、800μg·mL^(-1)黄芪多糖处理);将MDA-MB-453细胞分为MDA-MB-453空白组(常规培养)和MDA-MB-453低、中、高剂量实验组(分别给予200、400、800μg·mL^(-1)黄芪多糖处理)。以细胞计数试剂盒-8(CCK8)实验测定细胞增殖情况;以划痕实验检测细胞迁移情况;以Transwell法检测细胞侵袭情况;以蛋白质印迹法检测细胞β-连环蛋白(β-catenin)、c-myc、细胞周期素D1(Cyclin D1)蛋白表达水平。结果 BT-549细胞空白组和低、中、高剂量实验组的细胞存活率分别为(100.00±0.00)%、(91.24±3.61)%、(87.19±4.83)%和(84.45±2.66)%,迁移程度分别为(29.16±4.96)%、(25.69±4.14)%、(22.39±7.11)%和(11.25±1.30)%;MDA-MB-453细胞空白组和低、中、高剂量实验组的细胞存活率分别为(100.00±0.00)%、(99.10±4.57)%、(87.86±3.03)%和(82.58±4.60)%,迁移程度分别为(29.57±0.49)%、(22.00±5.04)%、(12.99±5.29)%和(2.95±0.20)%,中、高剂量实验组上述指标与空白组比较,差异均有统计学意义(均P<0.01)。BT-549细胞空白组和低、中、高剂量实验组的侵袭细胞数分别为304.00±15.10、90.50±13.03、63.50±3.11和34.25±14.01,低、中、高剂量实验组与空白组比较,差异均有统计学意义(均P<0.01)。BT-549细胞空白组和高剂量实验组β-catenin蛋白表达水平分别为1.00±0.00和0.52±0.15,c-myc蛋白表达水平分别为1.00±0.00和0.58±0.18,Cyclin D1蛋白表达水平分别为1.00±0.00和0.67±0.17,高剂量实验组上述指标与空白组比较,差异均有统计学意义(均P<0.01)。结论 黄芪多糖抑制三阴性乳腺癌细胞的增殖和侵袭,并调控Wnt/β-catenin通路。

Objective To explore the changes of proliferation and invasion ability of triple-negative breast cancer cells treated with Astragalus Polysaccharide (APS) and its influence on Wnt/β-catenin pathway.Methods BT-549 cells were divided into BT-549 contro group (conventional culture) and BT-549 experimental-L,-M,-H groups (treated with Astragalus Polysaccharide at 200,400 and 800μg·m L^(-1)respectively).MDA-MB-453 cells were divided into MDA-MB-453 control group (conventional culture) and MDA-MB-453experimental-L,-M,-H groups (treated with Astragalus Polysaccharide at 200,400 and 800μg·m L^(-1)respectively).Cell proliferation was measured by cell counting kit-8(CCK8);cell migration was detected by wound healing assay;cell invasion was detected by Transwell method;the expression levels of β-catenin,c-myc and Cyclin D1 were detected by Western blot.Results The survival rates of BT-549 cells in blank group,experimental-L,-M,-H groups were (100.00±0.00)%,(91.24±3.61)%,(87.19±4.83)%and (84.45±2.66)%,respectively;the migration indexes of each group were (29.16±4.96)%,(25.69±4.14)%,(22.39±7.11)%and (11.25±1.30)%,respectively;the survival rates of MDA-MB-453 cells in control group,experimental-L,-M,-H groups were (100.00±0.00)%,(99.10±4.57)%,(87.86±3.03)%and (82.58±4.60)%,respectively;the migration indexes were (29.57±0.49)%,(22.00±5.04)%,(12.99±5.29)%and(2.95±0.20)%,respectively.Compared with the control group,the above indexes in the experimental-M,-H groups have statistical significance (all P0.01).In the invasion assay of BT-549 cells,the number of invading cells were 304.00±15.10,90.50±13.03,63.50±3.11 and 34.25±14.01,respectively.Compared with the control group,the experimental-L,-M,-H groups had statistical significance (all P0.01).The expression levels of β-catenin,c-myc and Cyclin D1 protein in experimental-L,-M,-H groups of BT-549 cells were1.00±0.00 and 0.52±0.15,1.00±0.00 and 0.58±0.18,1.00±0.00 and 0.67±0.17,respectively.Compared with the control group,the differences in the above indexes in experimental-H group were statistically significant (all P0.01).Conclusion Astragalus Polysaccharide inhibits the proliferation and invasion of triple negative breast cancer cells and regulates Wnt/β-catenin pathway.