中华临床医师杂志(电子版)
中華臨床醫師雜誌(電子版)
중화림상의사잡지(전자판)
CHINESE JOURNAL OF CLINICIANS(ELECTRONIC VERSION)
2013年
23期
10798-10801
,共4页
商潇云%李鑫%于波%张素霜%杨秀静%李彦坤%曲欣%蔡朋朋%李洪源
商瀟雲%李鑫%于波%張素霜%楊秀靜%李彥坤%麯訢%蔡朋朋%李洪源
상소운%리흠%우파%장소상%양수정%리언곤%곡흔%채붕붕%리홍원
甘草%抗肿瘤药%细胞凋亡%HeLa细胞%PARP-1
甘草%抗腫瘤藥%細胞凋亡%HeLa細胞%PARP-1
감초%항종류약%세포조망%HeLa세포%PARP-1
Glycyrrhiza uralensis%Antineoplastic agents%Apoptosis%Hela cells%PARP-1?
目的:探讨甘草提取物(GC-3)抑制人宫颈癌HeLa细胞增殖及诱导HeLa细胞凋亡的机制。方法通过水提取、乙醇沉淀、大孔吸附树脂柱层析,分离提取GC-3;四甲基偶氮唑盐(MTT)比色法检测GC-3对HeLa细胞增殖的影响;透射电镜观察GC-3诱导HeLa细胞凋亡的形态学改变;Western blot测定25μg/ml GC-3不同时间作用于HeLa细胞后,凋亡相关蛋白聚腺苷二磷酸-核糖聚合酶(PARP)表达水平的变化。结果 MTT检测结果表明,GC-3能够抑制HeLa细胞增殖且呈现出良好的时间剂量效应关系,其中GC-3作用于人HeLa细胞12 h、24 h和48 h的半数抑制浓度(IC50)分别为35.21μg/ml、17.05μg/ml、12.07μg/ml,而对照组顺铂相对应的IC50分别为20.48μg/ml、7.34μg/ml、8.66μg/ml。电镜检测可见细胞表面绒毛脱失,细胞核内染色质凝聚、散在或边集,细胞的核浆比减少,胞质内可见空泡等明显的凋亡细胞特点。Western blot检测结果显示,随着GC-3(25μg/ml)作用时间的增加,PARP-1被激活剪切,其裂解蛋白Cleaved-PARP-124 kD表达水平随之增高,且从6 h时起即与正常对照组有显著性差异(P<0.05),呈现明显的时间效应关系。结论 GC-3在体外可明显抑制HeLa细胞的增殖和诱导HeLa细胞凋亡,PARP-1在该细胞分子凋亡机制中起重要作用。
目的:探討甘草提取物(GC-3)抑製人宮頸癌HeLa細胞增殖及誘導HeLa細胞凋亡的機製。方法通過水提取、乙醇沉澱、大孔吸附樹脂柱層析,分離提取GC-3;四甲基偶氮唑鹽(MTT)比色法檢測GC-3對HeLa細胞增殖的影響;透射電鏡觀察GC-3誘導HeLa細胞凋亡的形態學改變;Western blot測定25μg/ml GC-3不同時間作用于HeLa細胞後,凋亡相關蛋白聚腺苷二燐痠-覈糖聚閤酶(PARP)錶達水平的變化。結果 MTT檢測結果錶明,GC-3能夠抑製HeLa細胞增殖且呈現齣良好的時間劑量效應關繫,其中GC-3作用于人HeLa細胞12 h、24 h和48 h的半數抑製濃度(IC50)分彆為35.21μg/ml、17.05μg/ml、12.07μg/ml,而對照組順鉑相對應的IC50分彆為20.48μg/ml、7.34μg/ml、8.66μg/ml。電鏡檢測可見細胞錶麵絨毛脫失,細胞覈內染色質凝聚、散在或邊集,細胞的覈漿比減少,胞質內可見空泡等明顯的凋亡細胞特點。Western blot檢測結果顯示,隨著GC-3(25μg/ml)作用時間的增加,PARP-1被激活剪切,其裂解蛋白Cleaved-PARP-124 kD錶達水平隨之增高,且從6 h時起即與正常對照組有顯著性差異(P<0.05),呈現明顯的時間效應關繫。結論 GC-3在體外可明顯抑製HeLa細胞的增殖和誘導HeLa細胞凋亡,PARP-1在該細胞分子凋亡機製中起重要作用。
목적:탐토감초제취물(GC-3)억제인궁경암HeLa세포증식급유도HeLa세포조망적궤제。방법통과수제취、을순침정、대공흡부수지주층석,분리제취GC-3;사갑기우담서염(MTT)비색법검측GC-3대HeLa세포증식적영향;투사전경관찰GC-3유도HeLa세포조망적형태학개변;Western blot측정25μg/ml GC-3불동시간작용우HeLa세포후,조망상관단백취선감이린산-핵당취합매(PARP)표체수평적변화。결과 MTT검측결과표명,GC-3능구억제HeLa세포증식차정현출량호적시간제량효응관계,기중GC-3작용우인HeLa세포12 h、24 h화48 h적반수억제농도(IC50)분별위35.21μg/ml、17.05μg/ml、12.07μg/ml,이대조조순박상대응적IC50분별위20.48μg/ml、7.34μg/ml、8.66μg/ml。전경검측가견세포표면융모탈실,세포핵내염색질응취、산재혹변집,세포적핵장비감소,포질내가견공포등명현적조망세포특점。Western blot검측결과현시,수착GC-3(25μg/ml)작용시간적증가,PARP-1피격활전절,기렬해단백Cleaved-PARP-124 kD표체수평수지증고,차종6 h시기즉여정상대조조유현저성차이(P<0.05),정현명현적시간효응관계。결론 GC-3재체외가명현억제HeLa세포적증식화유도HeLa세포조망,PARP-1재해세포분자조망궤제중기중요작용。
Objective To investigate the apoptosis of human cervical cancer HeLa cells induced by extract of licorice in vitro and related molecular mechanism. Methods GC-3 was extracted by means of alcohol precipitation;Macroporous resin chromatography. 2MTT was used to evaluate the growth inhibition rate of HeLa cells which were treated by different concentrations of GC-3. Transmission electron microscopy (TEM) were used to observe the cells morphology. Western blot was used to detect the cell apoptosis related protein in Hela cells treated by GC-3 (25μg/ml) at different times. Results The anti-tumor active components (GC-3, extracted from licorice) showed time-dose-dependent inhibitory effect on the proliferation of HeLa cells in vitro. TEM detection showed that HeLa cells presented characteristic morphological changes of apoptosis after treated by GC-3. The results of Western blot revealed that after treated by GC-3 (25μg/ml) on HeLa cells at different times, GC-3 can promote the activation of PARP-1. Conclusion The anti-tumor active components (GC-3) extracted from licorice could inhibit the proliferation of HeLa cells and could also result in HeLa cell apoptosis in vitro, PARP-1 plays an important role in this apoptosis molecular mechanism.