红凉伞提取物调控MNX1-AS1对乳腺癌细胞增殖和凋亡的影响

Effect of Extract of Ardisia Crenata Var. Bicolor Regulating MNX1-AS1 on Proliferation and Apoptosis of Breast Cancer Cells

目的:探讨红凉伞提取物对乳腺癌细胞增殖和凋亡的影响及分子机制。方法:将乳腺癌细胞MDA-MB-453分为对照组、红凉伞提取物低、中、高剂量组、红凉伞提取物+pc DNA3.1组和红凉伞提取物+pc DNA3.1-MNX1-AS1组。四甲基偶氮唑盐比色法检测细胞增殖抑制率;克隆形成实验检测细胞克隆形成能力;流式细胞术检测细胞凋亡;Transwell检测细胞迁移和侵袭;实时荧光定量PCR检测MNX1-AS1表达水平;蛋白质印迹法检测细胞周期蛋白依赖性激酶抑制剂1A (P21)、半胱氨酸天冬氨酸蛋白酶-3 (Caspase-3)、上皮钙黏蛋白(E-cadherin)、基质金属蛋白酶-2 (MMP-2)表达水平。结果:与对照组比较,红凉伞提取物低、中、高剂量组细胞增殖抑制率、凋亡率及P21、Caspase-3、E-cadherin表达水平均升高(P<0.05),细胞克隆形成数、迁移、侵袭细胞数及MNX1-AS1、MMP-2表达水平均降低(P<0.05)。与红凉伞提取物+pc DNA3.1组比较,红凉伞提取物+pc DNA3.1-MNX1-AS1组细胞增殖抑制率、凋亡率及P21、Caspase-3、E-cadherin表达水平均降低(P<0.05),细胞克隆形成数、迁移、侵袭细胞数及MNX1-AS1、MMP-2表达水平均升高(P<0.05)。结论:红凉伞提取物可抑制乳腺癌细胞MDA-MB-453增殖、迁移和侵袭,促进细胞凋亡,其机制可能与MNX1-AS1有关。

Objective: To discuss the effect of extract of Ardisia crenata var. bicolor on proliferation and apoptosis of breast cancer cells and its molecular mechanism. Methods:Breast cancer cells MDA-MB-453 were divided into the control group,the groups of extract of Ardisia crenata var. bicolor of low,medium and high dose,the extract of Ardisia crenata var.bicolor +pcDNA3.1 group,and the extract of Ardisia crenata var. bicolor +pcDNA3.1-MNX1-AS1 group. The inhibition rate of cell proliferation was tested by methyl thiazolyl tetrazolium assay method. The colonly forming ability of cells was tested by colony formation assay. Cell apoptosis was detected by flow cytometry. Cell migration and invasion was detected by Transwell method. The expression level of MNX1-AS1 was detected by quantitative real-time PCR. The protein expression of cyclindependent kinase inhibitor 1 A(P21),cysteinyl aspartate specific proteinase-3(Caspase-3),Epithelial-cadherin(E-cadherin),and matrix metalloproteinase-2(MMP-2) was detected by Western blot method. Results: Compared with those in the control group,in the groups of extract of Ardisia crenata var. bicolor of low,medium and high dose,the inhibition rate of cell proliferation,the apoptosis rate,and the expression levels of P21,Caspase-3 and E-cadherin were increased(P<0.05).Compared with those in the control group,in the groups of extract of Ardisia crenata var. bicolor of low,medium and high dose,the number of cell clone formation,migration and invasion,as well as the expression levels of MNX1-AS1 and MMP-2 were decreased(P<0.05). Compared with those in the extract of Ardisia crenata var. bicolor + pcDNA3.1 group, in the extract of Ardisia crenata var. bicolor +pcDNA3.1-MNX1-AS1 group,the inhibition rate of cell proliferation,the apoptosis rate,and the expression levels of P21,Caspase-3 and E-cadherin were decreased(P<0.05). Compared with those in the extract of Ardisia crenata var. bicolor +pcDNA3.1 group,in the extract of Ardisia crenata var. bicolor +pcDNA3.1-MNX1-AS1 group, the number of cell clone formation, migration and invasion, as well as the expression levels of MNX1-AS1 and MMP-2 were increased(P<0.05). Conclusion:Extract of Ardisia crenata var. bicolor can inhibit proliferation,migration and invasion of breast cancer cells MDA-MB-453,and promote cell apoptosis. The mechanism may be related with MNX1-AS1.