世界科技研究与发展
世界科技研究與髮展
세계과기연구여발전
WORLD SCI-TECH R & D
2014年
3期
281-285
,共5页
鲁永婷%李惠新%拓振合%张栋栋%李瑞萍%郭钰珍
魯永婷%李惠新%拓振閤%張棟棟%李瑞萍%郭鈺珍
로영정%리혜신%탁진합%장동동%리서평%곽옥진
氧化苦参碱%SiHa细胞株%细胞增殖%细胞凋亡
氧化苦參堿%SiHa細胞株%細胞增殖%細胞凋亡
양화고삼감%SiHa세포주%세포증식%세포조망
oxymatrine%SiHa cells%cell proliferation%cell apoptosis
目的：研究氧化苦参碱对体外人宫颈癌SiHa细胞株增殖活性及凋亡的影响。方法对人宫颈癌SiHa细胞株进行体外培养，经氧化苦参碱处理的细胞组为实验组，未经处理组为对照组。采用MTT细胞存活实验检测氧化苦参碱对人宫颈鳞癌SiHa细胞的增殖影响，并计算IC50；倒置相差显微镜下观察不同浓度的氧化苦参碱作用48h后SiHa细胞的形态改变；Hoechst33258染色法和 Western blot法检测氧化苦参碱对 SiHa 细胞核及凋亡相关蛋白（P53、Bax及Bcl-2）表达水平的影响。结果 MTT结果显示氧化苦参碱剂量-时间依耐性抑制人宫颈癌SiHa细胞的体外增殖（P＜0.05），计算24 h、48 h和72 h的IC50分别为（1028.41±3.57）μg/ml、（701.72±6.01）μg/ml和（406.88±2.15）μg/ml；氧化苦参碱处理48 h后，SiHa细胞的形态特征及数目发生显著变化，且随氧化苦参碱浓度的增加而愈加明显；Hoechst33258染色证实氧化苦参碱处理后SiHa细胞发生凋亡，可见典型的凋亡特征；Western blot结果证实氧化苦参碱（700μg/ml、1600μg/ml）处理48 h后，和对照组比较，药物处理组细胞凋亡周期蛋白P53、Bax表达上调（P＜0.05），而Bcl-2表达下调（P＜0.05），Bax/Bcl-2的比率明显增加（P＜0.05）。结论氧化苦参碱可抑制体外人宫颈癌SiHa细胞的体外增殖活性发挥其抗肿瘤作用，其作用机制可能诱导SiHa细胞的凋亡有关。
目的：研究氧化苦參堿對體外人宮頸癌SiHa細胞株增殖活性及凋亡的影響。方法對人宮頸癌SiHa細胞株進行體外培養，經氧化苦參堿處理的細胞組為實驗組，未經處理組為對照組。採用MTT細胞存活實驗檢測氧化苦參堿對人宮頸鱗癌SiHa細胞的增殖影響，併計算IC50；倒置相差顯微鏡下觀察不同濃度的氧化苦參堿作用48h後SiHa細胞的形態改變；Hoechst33258染色法和 Western blot法檢測氧化苦參堿對 SiHa 細胞覈及凋亡相關蛋白（P53、Bax及Bcl-2）錶達水平的影響。結果 MTT結果顯示氧化苦參堿劑量-時間依耐性抑製人宮頸癌SiHa細胞的體外增殖（P＜0.05），計算24 h、48 h和72 h的IC50分彆為（1028.41±3.57）μg/ml、（701.72±6.01）μg/ml和（406.88±2.15）μg/ml；氧化苦參堿處理48 h後，SiHa細胞的形態特徵及數目髮生顯著變化，且隨氧化苦參堿濃度的增加而愈加明顯；Hoechst33258染色證實氧化苦參堿處理後SiHa細胞髮生凋亡，可見典型的凋亡特徵；Western blot結果證實氧化苦參堿（700μg/ml、1600μg/ml）處理48 h後，和對照組比較，藥物處理組細胞凋亡週期蛋白P53、Bax錶達上調（P＜0.05），而Bcl-2錶達下調（P＜0.05），Bax/Bcl-2的比率明顯增加（P＜0.05）。結論氧化苦參堿可抑製體外人宮頸癌SiHa細胞的體外增殖活性髮揮其抗腫瘤作用，其作用機製可能誘導SiHa細胞的凋亡有關。
목적：연구양화고삼감대체외인궁경암SiHa세포주증식활성급조망적영향。방법대인궁경암SiHa세포주진행체외배양，경양화고삼감처리적세포조위실험조，미경처리조위대조조。채용MTT세포존활실험검측양화고삼감대인궁경린암SiHa세포적증식영향，병계산IC50；도치상차현미경하관찰불동농도적양화고삼감작용48h후SiHa세포적형태개변；Hoechst33258염색법화 Western blot법검측양화고삼감대 SiHa 세포핵급조망상관단백（P53、Bax급Bcl-2）표체수평적영향。결과 MTT결과현시양화고삼감제량-시간의내성억제인궁경암SiHa세포적체외증식（P＜0.05），계산24 h、48 h화72 h적IC50분별위（1028.41±3.57）μg/ml、（701.72±6.01）μg/ml화（406.88±2.15）μg/ml；양화고삼감처리48 h후，SiHa세포적형태특정급수목발생현저변화，차수양화고삼감농도적증가이유가명현；Hoechst33258염색증실양화고삼감처리후SiHa세포발생조망，가견전형적조망특정；Western blot결과증실양화고삼감（700μg/ml、1600μg/ml）처리48 h후，화대조조비교，약물처리조세포조망주기단백P53、Bax표체상조（P＜0.05），이Bcl-2표체하조（P＜0.05），Bax/Bcl-2적비솔명현증가（P＜0.05）。결론양화고삼감가억제체외인궁경암SiHa세포적체외증식활성발휘기항종류작용，기작용궤제가능유도SiHa세포적조망유관。
Objective To evaluate the proliferative effect and induction of apoptosis by oxymatrine in human cervical cancer cell line SiHa in vitro.Methods SiHa were cultured in vitro.The experimental groups were treated with oxymatrine,the oth-ers were in the control groups.Proliferation rates and IC50 values were assessed by methyl thiazolyl tetrazolium (MTT).In-verted phase contrast microscope was used to observed the morphological alterations of SiHa human cervical cancer cells at five concentrations of oxymatrine at 48 h.The morphological changes accompanying cell apoptosis were observed using an inverted microscope after Hoechst 33258 staining.In addition,expression changes of apoptosis-related proteins involved in the P53,Bax and Bcl-2 were determined by western blotting.Results The results determined by MTT assay showed that the viability of human cervical cancer SiHa cells with oxymatrine was markedly attenuated in a concentration and time depend-ent manner(P<0.05).The IC50 values were calculated as (1 028.41 ±3.57)μg/ml,(701.72 ±6.01)μg/ml and (406.88 ±2.15)μg/ml at 24 h,48 h and 72 h.The morphological alterations of SiHa human cervical cancer cells changed significantly with the increase of the concentration of oxymatrine at 48 h.Also the oxymatrine-treated SiHa human cervical cancer cells displayed typical apoptosis morphological changes by the Hoechst 33258 staining at 48h.Compared with the control groups,the experimental groups resulted in remarkable increase of both P53 and Bax(P<0.05 ),but significantly decrease of Bcl-2(P<0.05)in Western blot.Conclusion These results demonstrate that oxymatrine exerts antitumor effects on SiHa cells through its anti-proliferative role,and suggest the mechanism for the effect may be related to its induction of apoptosis effects.
