临床肿瘤学杂志
臨床腫瘤學雜誌
림상종류학잡지
CHINESE CLINICAL ONCOLOGY
2015年
6期
481-486
,共6页
芒柄花黄素%非小细胞肺癌%细胞增殖%凋亡%上皮间质转化
芒柄花黃素%非小細胞肺癌%細胞增殖%凋亡%上皮間質轉化
망병화황소%비소세포폐암%세포증식%조망%상피간질전화
Formononetin%Non-small cell lung cancer%Cell proliferation%Apoptosis%Epithelial mesenchymal tran-sition( EMT)
目的：探讨芒柄花黄素（ Form）对人非小细胞肺癌A549细胞增殖、凋亡及上皮间质转化（ EMT）的影响。方法采用终浓度为0．1、1、10、100μmol／L Form处理A549细胞，应用四甲基偶氮唑盐比色法测定对照组（不加药物）及不同浓度Form处理24、48、72和96 h的细胞增殖情况，分别采用Annexin V?FITC／PI双染联合流式细胞术和实时荧光定量聚合酶链反应检测不同浓度Form处理48 h的凋亡率及凋亡相关基因的表达情况，Western blotting检测各组处理48 h后的EMT相关分子标志物（ E?cadherin、N?cadherin及Vimentin）水平，采用酶联免疫吸附法测各组处理24、48、72及96 h的细胞上清液中纤连蛋白（ FN）水平。结果 Form在1～100μmol／L的范围内对A549细胞有增殖抑制作用，增殖抑制率呈浓度和时间依赖性，且除24～48 h 0．1μmol／L外，其余浓度及作用时间的增殖抑制率均高于对照组（ P＜0．05）；与对照组比较，Form处理48 h后的早期、晚期凋亡率及Bax、Bak的mRNA水平均升高，而Bcl?2水平降低，差异均有统计学意义（ P＜0．05），且呈剂量依赖性；与对照组比较，不同浓度Form处理48 h后的N?cadherin和Vimentin水平及上清液FN水平均降低，而E?cadherin水平升高，以上差异有统计学意义（ P＜0．05）。结论 Form对非小细胞肺癌A549细胞有毒性作用，不仅抑制其增殖还诱导凋亡，可能与抑制其EMT过程有关。
目的：探討芒柄花黃素（ Form）對人非小細胞肺癌A549細胞增殖、凋亡及上皮間質轉化（ EMT）的影響。方法採用終濃度為0．1、1、10、100μmol／L Form處理A549細胞，應用四甲基偶氮唑鹽比色法測定對照組（不加藥物）及不同濃度Form處理24、48、72和96 h的細胞增殖情況，分彆採用Annexin V?FITC／PI雙染聯閤流式細胞術和實時熒光定量聚閤酶鏈反應檢測不同濃度Form處理48 h的凋亡率及凋亡相關基因的錶達情況，Western blotting檢測各組處理48 h後的EMT相關分子標誌物（ E?cadherin、N?cadherin及Vimentin）水平，採用酶聯免疫吸附法測各組處理24、48、72及96 h的細胞上清液中纖連蛋白（ FN）水平。結果 Form在1～100μmol／L的範圍內對A549細胞有增殖抑製作用，增殖抑製率呈濃度和時間依賴性，且除24～48 h 0．1μmol／L外，其餘濃度及作用時間的增殖抑製率均高于對照組（ P＜0．05）；與對照組比較，Form處理48 h後的早期、晚期凋亡率及Bax、Bak的mRNA水平均升高，而Bcl?2水平降低，差異均有統計學意義（ P＜0．05），且呈劑量依賴性；與對照組比較，不同濃度Form處理48 h後的N?cadherin和Vimentin水平及上清液FN水平均降低，而E?cadherin水平升高，以上差異有統計學意義（ P＜0．05）。結論 Form對非小細胞肺癌A549細胞有毒性作用，不僅抑製其增殖還誘導凋亡，可能與抑製其EMT過程有關。
목적：탐토망병화황소（ Form）대인비소세포폐암A549세포증식、조망급상피간질전화（ EMT）적영향。방법채용종농도위0．1、1、10、100μmol／L Form처리A549세포，응용사갑기우담서염비색법측정대조조（불가약물）급불동농도Form처리24、48、72화96 h적세포증식정황，분별채용Annexin V?FITC／PI쌍염연합류식세포술화실시형광정량취합매련반응검측불동농도Form처리48 h적조망솔급조망상관기인적표체정황，Western blotting검측각조처리48 h후적EMT상관분자표지물（ E?cadherin、N?cadherin급Vimentin）수평，채용매련면역흡부법측각조처리24、48、72급96 h적세포상청액중섬련단백（ FN）수평。결과 Form재1～100μmol／L적범위내대A549세포유증식억제작용，증식억제솔정농도화시간의뢰성，차제24～48 h 0．1μmol／L외，기여농도급작용시간적증식억제솔균고우대조조（ P＜0．05）；여대조조비교，Form처리48 h후적조기、만기조망솔급Bax、Bak적mRNA수평균승고，이Bcl?2수평강저，차이균유통계학의의（ P＜0．05），차정제량의뢰성；여대조조비교，불동농도Form처리48 h후적N?cadherin화Vimentin수평급상청액FN수평균강저，이E?cadherin수평승고，이상차이유통계학의의（ P＜0．05）。결론 Form대비소세포폐암A549세포유독성작용，불부억제기증식환유도조망，가능여억제기EMT과정유관。
Objective To explore the inhibitory effect of formononetin ( Form) on proliferation and epithelial mesenchymal tran?sition ( EMT) of non?small cell lung cancer A549 cells. Methods The A549 cells were cultured with Form ( a final concentration of 0. 1, 1, 10, 100μmol/L) . The cell proliferation was determined at 24, 48, 72 and 96 h post?treatment by MTT method in control group and From group. The Annexin V?FITC/PI double staining combined with flow cytometry and quantitative real?timepolymerase chain reac?tion were applied to measure the early?apoptotic and late?apoptotic rates and expression of apoptosis related genes at 48 h after treatment. Western blotting was used to detect the EMT related molecular markers, including E?cadherin, N?cadherin and Vimentin. The supernatant levels of fibronectin ( FN) were measured at 24, 48, 72 and 96 h by enzyme linked immunosorbent assay. Results Form exerted the in?hibitory effect on the proliferation of A549 cells in the range of 1?100 mol/L, and presented in a dose?and time?dependent manner. In ad?dition to 0. 1 mol/L at 24?48 h, the proliferation inhibiting rates of Form were higher than those of control group ( P<0. 05) . Compared with the control group, there were higher levels of early?apoptotic and late?apoptotic rates and mRNA levels of Bax and Bak but lower Bcl?2 mRNA levels in Form groups with statistical significance (P<0. 05). In response to Form, there were increased E?cadherin but de?creased N?cadherin, Vimentin and FN in comparison with control group at 48 h. Conclusion Form has a toxic effect on A549, which not only inhibit the proliferation and induce apoptosis. It may be related to the inhibition of the EMT process.
