中国临床药理学杂志
中國臨床藥理學雜誌
중국림상약이학잡지
THE CHINESE JOURNAL OF CLINICAL PHARMACOLOGY
2015年
11期
938-941
,共4页
成志勇%颜晓燕%李琳%谢旭磊%王凤云%李继业%王素云
成誌勇%顏曉燕%李琳%謝旭磊%王鳳雲%李繼業%王素雲
성지용%안효연%리림%사욱뢰%왕봉운%리계업%왕소운
青蒿琥酯%K562细胞系%10号染色体缺失的磷酸酶基因%凋亡
青蒿琥酯%K562細胞繫%10號染色體缺失的燐痠酶基因%凋亡
청호호지%K562세포계%10호염색체결실적린산매기인%조망
Artesunate%K562 cell line%phosphatase and tensin hemology deleted on chromosome ten gene%apoptosis
目的:探讨青蒿琥酯经10号染色体缺失的磷酸酶及张力蛋白同源基因( PTEN)介导的信号通路诱导人白血病K562细胞凋亡作用机制。方法将5个浓度(0,6.25,12.5,25,50μg· mL-1)浓度的青蒿琥酯作用K562细胞0-72 h,通过噻唑蓝比色法观察其对K562细胞生长抑制作用;罗丹明123检测线粒体膜电位变化;荧光定量聚合酶链反应检测PTEN mRNA水平变化,试剂盒检测半胱天冬酶(Caspase)3/7活性;蛋白印迹法检测PTEN、蛋白激酶B(p-Akt)蛋白水平。结果青蒿琥酯能显著抑制K562细胞增殖,72 h最大抑制率为91.5%,与剂量和时间呈正相关。青蒿琥酯明显降低K562细胞线粒体膜电位,增加PTEN表达,抑制p-Akt表达,增强Caspase3/7蛋白活性。结论青蒿琥酯可能通过PTEN/Akt/Caspase通路诱导K562细胞凋亡。
目的:探討青蒿琥酯經10號染色體缺失的燐痠酶及張力蛋白同源基因( PTEN)介導的信號通路誘導人白血病K562細胞凋亡作用機製。方法將5箇濃度(0,6.25,12.5,25,50μg· mL-1)濃度的青蒿琥酯作用K562細胞0-72 h,通過噻唑藍比色法觀察其對K562細胞生長抑製作用;囉丹明123檢測線粒體膜電位變化;熒光定量聚閤酶鏈反應檢測PTEN mRNA水平變化,試劑盒檢測半胱天鼕酶(Caspase)3/7活性;蛋白印跡法檢測PTEN、蛋白激酶B(p-Akt)蛋白水平。結果青蒿琥酯能顯著抑製K562細胞增殖,72 h最大抑製率為91.5%,與劑量和時間呈正相關。青蒿琥酯明顯降低K562細胞線粒體膜電位,增加PTEN錶達,抑製p-Akt錶達,增彊Caspase3/7蛋白活性。結論青蒿琥酯可能通過PTEN/Akt/Caspase通路誘導K562細胞凋亡。
목적:탐토청호호지경10호염색체결실적린산매급장력단백동원기인( PTEN)개도적신호통로유도인백혈병K562세포조망작용궤제。방법장5개농도(0,6.25,12.5,25,50μg· mL-1)농도적청호호지작용K562세포0-72 h,통과새서람비색법관찰기대K562세포생장억제작용;라단명123검측선립체막전위변화;형광정량취합매련반응검측PTEN mRNA수평변화,시제합검측반광천동매(Caspase)3/7활성;단백인적법검측PTEN、단백격매B(p-Akt)단백수평。결과청호호지능현저억제K562세포증식,72 h최대억제솔위91.5%,여제량화시간정정상관。청호호지명현강저K562세포선립체막전위,증가PTEN표체,억제p-Akt표체,증강Caspase3/7단백활성。결론청호호지가능통과PTEN/Akt/Caspase통로유도K562세포조망。
Objective To investigate the effective mechanism of Artesu-nate inducing apoptosis in human leukemia K562 cells through phospha-tase and tensin hemology deleted on chromosome ten gene ( PTEN) path-way.Methods The K562 cells were treated with concentrations of 0 , 6.25,12.5,25,50μg.mL-1 Artesunate within 0-72 hours.The growth inhibition of Artesunate on K562 cells was detected by thiazole blue assay.The mitochondrial membrane potential was assessed by rhodamine 123 with flow cytometry.The PTEN mRNA expression levels were detec-ted by real-time fluorescent quantitative reverse transcription polymerase chain reaction.Cysteine aspartic acid specific protease ( Caspase ) -3/7 protein activitives were tested by kits.PTEN and protein kinase B ( p-Akt) protein levels weres detected by Western Blotting.Results Arte-sunate has significant proliferation inhibition effect on K562 cells.The maximum growth inhibition ratio was 91.5% in 72 hours and the anti-proliferative effect was in a time and dose dependent manner.The mito-chondrial membrane potential was significantly decreased.PTEN was up regulated.The p-Akt was down regulated and caspase-3/7 protein ac-tivity was enhanced by Artesunate.Conclusion Artesunate possiblely induced K562 cells apoptosis via PTEN/Akt/Caspase pathway.