黄芩苷对多发性骨髓瘤细胞株U266增殖、凋亡、侵袭的影响  

Effects of baicalin on proliferation,apoptosis,and invasion of multiple myeloma U266 cells

作　　者：赵含笑 于天启 ZHAO Hanxiao;YU Tianqi(The Third Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510378,China)

机构地区：[1]广州中医药大学第三附属医院,广州510378

出　　处：《山东医药》2018年第47期28-31,共4页Shandong Medical Journal

基　　金：广东省自然科学基金项目(2014A020212272)

摘　　要：目的观察黄芩苷对多发性骨髓瘤细胞株U266增殖、凋亡、侵袭的影响,并探讨其作用机制。方法将U266细胞分为4组,空白组加入不含药物的培养基、地塞米松组加入地塞米松、黄芩苷组加入黄芩苷、地塞米松+黄芩苷组加入地塞米松和黄芩苷,分别培养。培养24、48 h时,CCK-8法检测各组OD值,以OD值反映各组细胞增殖抑制情况;培养24 h时流式细胞术Annexin V-FITC/PI双染法测算各组细胞凋亡率;培养12 h时Transwell侵袭实验检测各组穿膜细胞数,以穿膜细胞数表示细胞侵袭能力; RT-PCR法检测各组细胞Wnt通路调节分子β-catenin、GSK3β及下游靶基因c-Myc、cyclin D1 mRNA; Western blotting法检测各组细胞β-catenin、GSK3β蛋白。结果培养24 h时,空白组、地塞米松组、黄芩苷组、地塞米松+黄芩苷组的OD值分别为0. 33±0. 01、0. 29±0. 01、0. 30±0. 01、0. 27±0. 02,地塞米松+黄芩苷组与空白组相比,P <0. 05;培养48 h时,空白组、地塞米松组、黄芩苷组、地塞米松+黄芩苷组的OD值分别为0. 57±0. 02、0. 42±0. 02、0. 49±0. 01、0. 36±0. 02,组间相比,P均<0. 05。培养24 h时,空白组、地塞米松组、黄芩苷组、地塞米松+黄芩苷组细胞凋亡率分别为5. 4%、17. 8%、13. 8%、30. 5%,黄芩苷组与地塞米松组相比,P> 0. 05;其余各组间相比,P均<0. 05。培养12 h时,空白组、地塞米松组、黄芩苷组、地塞米松+黄芩苷组每个镜下穿膜细胞数分别为(99±8. 19)、(88. 33±6. 11)、(87. 67±2. 08)、(32. 67±3. 5)个,黄芩苷组与地塞米松组相比,P> 0. 05;其余各组间相比,P均<0. 05。与空白组相比,黄芩苷组、地塞米松组、地塞米松+黄芩苷组β-catenin、GSK3β、c-Myc、cyclin D1 mRNA的表达均下调(P均<0. 05);与空白组相比,地塞米松+黄芩苷组β-catenin、GSK3β蛋白的表达均下调(P均<0. 05)。结论黄芩苷可抑制多发性骨髓瘤细胞的增殖、侵袭,促进其凋亡,作用效果与地塞米松类似;黄岑Objective To observe the effects of baicalin on proliferation,apoptosis,and invasion of multiple myeloma U266 cells and its mechanism.Methods U266 cells were divided into 4 groups.The blank group was added with drug-free medium,dexamethasone group was added with dexamethasone,baicalin group was added with baicalin,and the dexamethasone+baicalin group were added with dexamethasone and baicalin.At 24 and 48 h of culture,CCK-8 was used to detect the OD value of each group(the OD value was used to reflect the inhibition of cell proliferation in each group).When cultured for 24 h,the apoptosis rate of cells in each group was measured by flow cytometry Annexin V-FITC/PI double staining.Transwell invasion assay was used to detect the number of transmembrane cells in each group(the cell invasion ability was expressed by the number of transmembrane cells).RT-PCR was used to detect the expression of Wnt pathway regulatory moleculesβ-catenin,GSK3βand downstream target genes c-Myc and cyclin D1 mRNA in each group.Western blotting was used to detectβ-catenin and GSK3βproteins in each group.Results At 24 h,the OD values of the blank group,dexamethasone group,baicalin group,and the dexamethasone+baicalin group were 0.33±0.01,0.29±0.01,0.30±0.01,and 0.27±0.02,respectively;significant difference was found between the dexamethasone+baicalin group and blank group(P<0.05),but no difference was found between the other groups(all P<0.05).At 48 h,the OD values were 0.57±0.02,0.42±0.02,0.49±0.01,and 0.36±0.02,respectively,with statistically significant difference(all P<0.05).At 24h,the apoptosis rates of the blank group,dexamethasone group,baicalin group,and dexamethasone+baicalin group were 5.4%,17.8%,13.8%,and 30.5%,respectively;no significant difference was found between the dexamethasone group and baicalin group(P<0.05),but significant difference was found between the other groups(all P<0.05).At 12 h,the number of transmembrane cells per panel in the blank group,dexamethasone group,baicalin group,and dexamethasone+baical

关 键 词：黄芩苷 多发性骨髓瘤 地塞米松 WNT通路 

分 类 号：R273[医药卫生—中西医结合]