中华神经医学杂志
中華神經醫學雜誌
중화신경의학잡지
CHINESE JOURNAL OF NEUROMEDICINE
2014年
7期
654-658
,共5页
神经胶质瘤%白花丹素%SOX-2%miR200b%miR200c%miR-203%miR-21
神經膠質瘤%白花丹素%SOX-2%miR200b%miR200c%miR-203%miR-21
신경효질류%백화단소%SOX-2%miR200b%miR200c%miR-203%miR-21
Glioma%Plumbagin%SOX-2%MiR200b%MiR200c%MiR-203%MiR21
目的 分析中药提纯制剂白花丹素对胶质瘤细胞增殖、凋亡和侵袭能力的影响及其作用机制. 方法 体外常规培养SWO-38和U251细胞,应用0~50 μmol/L白花丹素作用48 h后,MTT检测细胞存活率的变化,计算半数抑制浓度(IC50);应用0、2.5、5、10 μmol/L白花丹素处理细胞48 h后,流式细胞仪检测细胞凋亡;划痕实验检测细胞侵袭迁移能力;Westem blotting检测细胞中SOX2蛋白的表达;实时荧光定量PCR检测细胞miR200b、miR200c、miR-203和miR-21表达的变化. 结果 MTT显示0~50 μmol/L白花丹素处理SWO-38、U251细胞后细胞存活率逐渐下降,IC50分别为6.8 μmol/L和7.4 μmol/L;0、2.5、5、10 μmol/L白花丹素作用SWO-38、U251细胞48h后凋亡细胞的比例逐渐增加,细胞中miR200b、miR200c和miR-203的表达逐渐增加,差异有统计学意义(p<0.05);划痕实验显示白花丹素作用后两株细胞的侵袭能力下降;Westem blotting结果显示SOX2蛋白的表达降低. 结论 白花丹素抑制胶质瘤细胞的增殖和侵袭迁移,促进其凋亡,可能与其升高miR200b、miR200c和miR-203的表达,降低SOX2蛋白的表达有关.
目的 分析中藥提純製劑白花丹素對膠質瘤細胞增殖、凋亡和侵襲能力的影響及其作用機製. 方法 體外常規培養SWO-38和U251細胞,應用0~50 μmol/L白花丹素作用48 h後,MTT檢測細胞存活率的變化,計算半數抑製濃度(IC50);應用0、2.5、5、10 μmol/L白花丹素處理細胞48 h後,流式細胞儀檢測細胞凋亡;劃痕實驗檢測細胞侵襲遷移能力;Westem blotting檢測細胞中SOX2蛋白的錶達;實時熒光定量PCR檢測細胞miR200b、miR200c、miR-203和miR-21錶達的變化. 結果 MTT顯示0~50 μmol/L白花丹素處理SWO-38、U251細胞後細胞存活率逐漸下降,IC50分彆為6.8 μmol/L和7.4 μmol/L;0、2.5、5、10 μmol/L白花丹素作用SWO-38、U251細胞48h後凋亡細胞的比例逐漸增加,細胞中miR200b、miR200c和miR-203的錶達逐漸增加,差異有統計學意義(p<0.05);劃痕實驗顯示白花丹素作用後兩株細胞的侵襲能力下降;Westem blotting結果顯示SOX2蛋白的錶達降低. 結論 白花丹素抑製膠質瘤細胞的增殖和侵襲遷移,促進其凋亡,可能與其升高miR200b、miR200c和miR-203的錶達,降低SOX2蛋白的錶達有關.
목적 분석중약제순제제백화단소대효질류세포증식、조망화침습능력적영향급기작용궤제. 방법 체외상규배양SWO-38화U251세포,응용0~50 μmol/L백화단소작용48 h후,MTT검측세포존활솔적변화,계산반수억제농도(IC50);응용0、2.5、5、10 μmol/L백화단소처리세포48 h후,류식세포의검측세포조망;화흔실험검측세포침습천이능력;Westem blotting검측세포중SOX2단백적표체;실시형광정량PCR검측세포miR200b、miR200c、miR-203화miR-21표체적변화. 결과 MTT현시0~50 μmol/L백화단소처리SWO-38、U251세포후세포존활솔축점하강,IC50분별위6.8 μmol/L화7.4 μmol/L;0、2.5、5、10 μmol/L백화단소작용SWO-38、U251세포48h후조망세포적비례축점증가,세포중miR200b、miR200c화miR-203적표체축점증가,차이유통계학의의(p<0.05);화흔실험현시백화단소작용후량주세포적침습능력하강;Westem blotting결과현시SOX2단백적표체강저. 결론 백화단소억제효질류세포적증식화침습천이,촉진기조망,가능여기승고miR200b、miR200c화miR-203적표체,강저SOX2단백적표체유관.
Objective To detect the effect of plumbagin on proliferation,apoptosis and invasion ofglioma cell lines,and investigate the underlying mechanism.Methods Glioma cell lines SWO-38 and U251 were routinely cultured in vitro,and treated with various concentrations of plumbagin (0-50 μmol/L) for 48 h; cell viability changes were detected by MTT assay,and median inhibitory concentration (IC50) was calculated; after 0,2.5,5 and 10 μmol/L plumbagin treatment for 48 h,cell apoptosis was detected by annexin V/propidium iodide double-labeled flow cytometry; scratch test was used to observe the cell invasion and migration; Western blotting was used to assess the SOX2 protein expression; MiR200b,miR200c,miR-203 and miR-21 expression changes were examined by real-time quantitative PCR.Results Plumbagin dose-dependently inhibited the proliferation of the glioma cells;the IC50 values ofplumbagin in SWO-38 and U251 cells were 6.8 and 7.4 μmol/L,respectively.After 0,2.5,5 and 10 μmol/L plumbagin treatment for 48 h,cell apoptosis ratio was gradually increased,and MiR200b,miR200c,miR-203 expressions gradually increased,with significant differences (P<0.05).Cell invasion and migration in these two cell lines were decreased and the SOX2 protein expression was decreased.Conclusion Plumbagin inhibits cell growth,induces apoptosis,and decrease cell invasion and migration,which might be related to the increase ofmiR200b,miR200c and miR-203 expressions and the decrease of SOX2 protein expression.