中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
Chinese Journal of Experimental Surgery
2015年
11期
2705-2707
,共3页
木犀草素%结肠癌%线粒体
木犀草素%結腸癌%線粒體
목서초소%결장암%선립체
Luteolin%Colorectal cancer%Mitochondria
目的 探讨木犀草素在体外诱导结肠癌HT-29细胞凋亡的作用机制.方法 体外培养人结肠癌细胞HT-29,以不同浓度木犀草素处理后,应用噻唑蓝(MTT)法检测木犀草素对HT-29细胞增殖的抑制作用;膜联蛋白V-异硫氰酸荧光素(Annexin V-FITC)/碘化丙锭(PI)双标记流式细胞术检测细胞凋亡率;采用实时定量聚合酶链反应(Real-time PCR)检测B细胞淋巴瘤/白血病-2(bcl-2)、B细胞淋巴瘤/白血病-2相关X蛋白(bax)、半胱氨酰天冬氨酸特异性蛋白酶(Caspase)-3、Caspase-9 mRNA表达水平;Western blot检测木犀草素对HT-29细胞内bcl-2、bax、Caspase-3、Caspase-9等蛋白表达的影响.结果 MTT结果显示木犀草素能显著抑制HT-29细胞的生长、增殖,呈浓度和时间效应关系(72 h最高抑制率达78.72％);流式细胞仪分析示木犀草素能诱导细胞发生凋亡,凋亡率可达(23.78±1.03)％;Real-time PCR检测结果示木犀草素上调了bax、Caspase-3、Caspase-9 mRNA表达,抑制了bcl-2 mRNA表达;木犀草素干预后,Western blot结果示bcl-2蛋白呈浓度剂量依赖性递减;而bax、Caspase-9、Caspase-3蛋白表达则显著增高.结论 木犀草素能通过线粒体通路抑制人结肠癌HT-29细胞的增殖并诱导其凋亡.
目的 探討木犀草素在體外誘導結腸癌HT-29細胞凋亡的作用機製.方法 體外培養人結腸癌細胞HT-29,以不同濃度木犀草素處理後,應用噻唑藍(MTT)法檢測木犀草素對HT-29細胞增殖的抑製作用;膜聯蛋白V-異硫氰痠熒光素(Annexin V-FITC)/碘化丙錠(PI)雙標記流式細胞術檢測細胞凋亡率;採用實時定量聚閤酶鏈反應(Real-time PCR)檢測B細胞淋巴瘤/白血病-2(bcl-2)、B細胞淋巴瘤/白血病-2相關X蛋白(bax)、半胱氨酰天鼕氨痠特異性蛋白酶(Caspase)-3、Caspase-9 mRNA錶達水平;Western blot檢測木犀草素對HT-29細胞內bcl-2、bax、Caspase-3、Caspase-9等蛋白錶達的影響.結果 MTT結果顯示木犀草素能顯著抑製HT-29細胞的生長、增殖,呈濃度和時間效應關繫(72 h最高抑製率達78.72％);流式細胞儀分析示木犀草素能誘導細胞髮生凋亡,凋亡率可達(23.78±1.03)％;Real-time PCR檢測結果示木犀草素上調瞭bax、Caspase-3、Caspase-9 mRNA錶達,抑製瞭bcl-2 mRNA錶達;木犀草素榦預後,Western blot結果示bcl-2蛋白呈濃度劑量依賴性遞減;而bax、Caspase-9、Caspase-3蛋白錶達則顯著增高.結論 木犀草素能通過線粒體通路抑製人結腸癌HT-29細胞的增殖併誘導其凋亡.
목적 탐토목서초소재체외유도결장암HT-29세포조망적작용궤제.방법 체외배양인결장암세포HT-29,이불동농도목서초소처리후,응용새서람(MTT)법검측목서초소대HT-29세포증식적억제작용;막련단백V-이류청산형광소(Annexin V-FITC)/전화병정(PI)쌍표기류식세포술검측세포조망솔;채용실시정량취합매련반응(Real-time PCR)검측B세포림파류/백혈병-2(bcl-2)、B세포림파류/백혈병-2상관X단백(bax)、반광안선천동안산특이성단백매(Caspase)-3、Caspase-9 mRNA표체수평;Western blot검측목서초소대HT-29세포내bcl-2、bax、Caspase-3、Caspase-9등단백표체적영향.결과 MTT결과현시목서초소능현저억제HT-29세포적생장、증식,정농도화시간효응관계(72 h최고억제솔체78.72％);류식세포의분석시목서초소능유도세포발생조망,조망솔가체(23.78±1.03)％;Real-time PCR검측결과시목서초소상조료bax、Caspase-3、Caspase-9 mRNA표체,억제료bcl-2 mRNA표체;목서초소간예후,Western blot결과시bcl-2단백정농도제량의뢰성체감;이bax、Caspase-9、Caspase-3단백표체칙현저증고.결론 목서초소능통과선립체통로억제인결장암HT-29세포적증식병유도기조망.
Objective To explore the action mechanism of Luteolin-induced apoptosis of human colorectal carcinoma HT-29 cells.Methods HT-29 cells were cultured in vitro, and treated with Luteolin at different concentrations.The inhibitory effects of Luteolin on cell proliferation was examined by methyl thiazol tetrazolium (MTT) assay, and the inhibition rate of 80 μmol/L Luteolin for 72 h was 78.72％.The cell apoptosis rate was detected by Annexin V-fluoresceine isothiocyanate (FITC)/propidium iodide (PI) double staining, and the apoptosis rate of HT-29 cells treated with Luteolin for 24 h was (23.78 ± 1.03) ％.The mRNA and protein levels of B cell lymphoma/leukemia-2 associated X protein (bax), B cell lymphoma/leukemia-2 (bcl-2), Caspase-3 and Caspase-9 were detected by real-time quantitative polymerase chain reaction (Real-time PCR) and Western blotting respectively.Results MTT assay showed Luteolin could inhibit the growth and proliferation of HT-29 cells in a time-and concentration-dependent manner.Flow cytometry showed that Luteolin could induce apoptosis of HT-29 cells.Luteolin could activate the expression of bax and Caspase-3,-9, and inhibit the expression of bcl-2.Conclusion Luteolin can significantly inhibit the proliferation and induce the apoptosis of HT-29 cells, which may be associated with mitochondrial pathway.
