江西中医药大学学报
江西中醫藥大學學報
강서중의약대학학보
Journal of Jiangxi University of Traditional Chinese Medicine
2014年
3期
82-84,100
,共4页
蛇床子素%Hela细胞%凋亡
蛇床子素%Hela細胞%凋亡
사상자소%Hela세포%조망
Osthole%Hela cell%apoptosis
目的：探讨蛇床子素诱导人宫颈癌Hela细胞凋亡及其机制。方法：采用MTT法检测各浓度蛇床子素对Hela细胞增殖的抑制作用；光学显微镜观察细胞的形态学变化；凝胶电泳分析DNA的片段化改变；采用RT-PCR检测Hela细胞fas和bcl-2基因mRNA的表达水平。采用10μg/mL的蛇床子素作用于Hela细胞24h后的培养上清称为ost-La；洗去蛇床子素，用新鲜培养液进行24 h再培养的Hela细胞培养上清称为ost-Lb；不经蛇床子素处理的Hela细胞同步培养上清作为对照，称为con-trol-L；定量Elisa法测定各上清中TGF-β1含量。结果：蛇床子素可显著抑制Hela细胞的体外增殖，诱导Hela细胞的凋亡，且均呈剂量依赖性。 RT-PCR：凋亡相关基因fas的表达量升高，而bcl-2的表达量下降，表达量的变化与蛇床子素的浓度有关。 Elisa：TGF-β1的含量在ost-La中为16．732±3．068 ng/mL（P＜0．01）；在ost-Lb中为2．031±0．112 ng/mL（P＜0．01）；在control-L中为385．282±42．613 ng/mL。结论：子素可诱导Hela细胞的凋亡，其机制可能与促进Fas基因表达，抑制Bcl-2基因表达和TGF-β1含量改变有关。
目的：探討蛇床子素誘導人宮頸癌Hela細胞凋亡及其機製。方法：採用MTT法檢測各濃度蛇床子素對Hela細胞增殖的抑製作用；光學顯微鏡觀察細胞的形態學變化；凝膠電泳分析DNA的片段化改變；採用RT-PCR檢測Hela細胞fas和bcl-2基因mRNA的錶達水平。採用10μg/mL的蛇床子素作用于Hela細胞24h後的培養上清稱為ost-La；洗去蛇床子素，用新鮮培養液進行24 h再培養的Hela細胞培養上清稱為ost-Lb；不經蛇床子素處理的Hela細胞同步培養上清作為對照，稱為con-trol-L；定量Elisa法測定各上清中TGF-β1含量。結果：蛇床子素可顯著抑製Hela細胞的體外增殖，誘導Hela細胞的凋亡，且均呈劑量依賴性。 RT-PCR：凋亡相關基因fas的錶達量升高，而bcl-2的錶達量下降，錶達量的變化與蛇床子素的濃度有關。 Elisa：TGF-β1的含量在ost-La中為16．732±3．068 ng/mL（P＜0．01）；在ost-Lb中為2．031±0．112 ng/mL（P＜0．01）；在control-L中為385．282±42．613 ng/mL。結論：子素可誘導Hela細胞的凋亡，其機製可能與促進Fas基因錶達，抑製Bcl-2基因錶達和TGF-β1含量改變有關。
목적：탐토사상자소유도인궁경암Hela세포조망급기궤제。방법：채용MTT법검측각농도사상자소대Hela세포증식적억제작용；광학현미경관찰세포적형태학변화；응효전영분석DNA적편단화개변；채용RT-PCR검측Hela세포fas화bcl-2기인mRNA적표체수평。채용10μg/mL적사상자소작용우Hela세포24h후적배양상청칭위ost-La；세거사상자소，용신선배양액진행24 h재배양적Hela세포배양상청칭위ost-Lb；불경사상자소처리적Hela세포동보배양상청작위대조，칭위con-trol-L；정량Elisa법측정각상청중TGF-β1함량。결과：사상자소가현저억제Hela세포적체외증식，유도Hela세포적조망，차균정제량의뢰성。 RT-PCR：조망상관기인fas적표체량승고，이bcl-2적표체량하강，표체량적변화여사상자소적농도유관。 Elisa：TGF-β1적함량재ost-La중위16．732±3．068 ng/mL（P＜0．01）；재ost-Lb중위2．031±0．112 ng/mL（P＜0．01）；재control-L중위385．282±42．613 ng/mL。결론：자소가유도Hela세포적조망，기궤제가능여촉진Fas기인표체，억제Bcl-2기인표체화TGF-β1함량개변유관。
Objective:To investigate the effects of Osthole on the apoptosis of Hela cells in vitro and to research its mechanism in detail . Methods:The inhibitory effects of Osthole on proliferation of Hela cells were detected using the MTT assay .After Hela cells were trea-ted with different doses of Osthole ,cell morphological alteration was observed by using optical microscopy , the DNA Fragmentation was examined by gel electrophoresis and RT -PCR was performed to measure the expression level of fas and bcl -2 mRNA.The superna-tant of Hela cells after treated with 10μg /mL Osthole respectively for 24 hours was called ost -La, and those followed by washing com-pletely to remove Osthole and re -cultured with fresh medium for 24 hours was called ost-Lb .The corresponding supernatants of He-la cells treated without Osthole is control -L.The concentration of TGF -β1 in different supernatant we measured by quantitative ELISA .Results:Osthole was found to inhibit proliferation and to induce the apoptosis of Hela cells in a concentration -dependent manner .RT-PCR:The expression of apoptosis -associated gene fas was up regulated , and the expression of bcl -2 was down regula-ted .The effects were dependent on concentration of Osthole .ELISA:The TGF-β1 content of ost -La was 16.732 ±3.068 ng/mL (P<0.01), of ost-Lb was 2.031 ±0.112ng/mL(P<0.01),of control-L was (385.282 ±42.613)ng/mL.Conclusion:Osthole can induce apoptosis of Hela cells .Its mechanism might relate to promoting the expression of fas , inhibiting the expression of bcl -2 and changing of the TGF-β1 content.
