中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2012年
9期
1759-1762
,共4页
许隽颖%杨洁%陈敏斌%李江%王润洁%陆培华
許雋穎%楊潔%陳敏斌%李江%王潤潔%陸培華
허준영%양길%진민빈%리강%왕윤길%륙배화
腺苷酸活化蛋白激酶%冬凌草甲素%结肠癌
腺苷痠活化蛋白激酶%鼕凌草甲素%結腸癌
선감산활화단백격매%동릉초갑소%결장암
AMP activated protein kinase%Oridonin%Colon carcinoma
目的 观察腺苷酸活化蛋白激酶(AMPK)在冬凌草甲素体外诱导结肠癌HT-29细胞凋亡中的作用.方法 浓度为1 ～ 50 μmol/L的冬凌草甲素分别作用结肠癌HT-29细胞,组蛋白-DNA酶联免疫吸附试验(ELISA)法检测冬凌草甲素诱导的结肠癌细胞凋亡率.分光光度法检测作用后的结肠癌细胞半胱氨酰天冬氨酸特异性蛋白酶( Caspase)-3活性,Western blot法测定冬凌草甲素作用的肿瘤细胞AMPK及其他凋亡相关蛋白表达.结果 不同浓度冬凌草甲素作用于结肠癌细胞后,结肠癌细胞发生凋亡.随着药物浓度增加,细胞凋亡率也逐渐增加(P＜0.05),且结肠癌细胞Caspase-3的活性也逐渐增加(P＜0.05).随着作用时间增加,肿瘤细胞p-AMPKα蛋白条带逐渐变深变粗.冬凌草甲素作用的转染AMPKα小干扰RNA(siRNA)的细胞凋亡率(26.33±5.03)％低于转染错义siRNA的细胞凋亡率(84.40 ±9.70)％,差异有统计学意义(P＜0.05).结论 冬凌草甲素诱导结肠癌HT-29细胞凋亡,活化AMPK直接参与其诱导的肿瘤细胞凋亡,其机制可能与Caspase-3的活性表达有关.
目的 觀察腺苷痠活化蛋白激酶(AMPK)在鼕凌草甲素體外誘導結腸癌HT-29細胞凋亡中的作用.方法 濃度為1 ～ 50 μmol/L的鼕凌草甲素分彆作用結腸癌HT-29細胞,組蛋白-DNA酶聯免疫吸附試驗(ELISA)法檢測鼕凌草甲素誘導的結腸癌細胞凋亡率.分光光度法檢測作用後的結腸癌細胞半胱氨酰天鼕氨痠特異性蛋白酶( Caspase)-3活性,Western blot法測定鼕凌草甲素作用的腫瘤細胞AMPK及其他凋亡相關蛋白錶達.結果 不同濃度鼕凌草甲素作用于結腸癌細胞後,結腸癌細胞髮生凋亡.隨著藥物濃度增加,細胞凋亡率也逐漸增加(P＜0.05),且結腸癌細胞Caspase-3的活性也逐漸增加(P＜0.05).隨著作用時間增加,腫瘤細胞p-AMPKα蛋白條帶逐漸變深變粗.鼕凌草甲素作用的轉染AMPKα小榦擾RNA(siRNA)的細胞凋亡率(26.33±5.03)％低于轉染錯義siRNA的細胞凋亡率(84.40 ±9.70)％,差異有統計學意義(P＜0.05).結論 鼕凌草甲素誘導結腸癌HT-29細胞凋亡,活化AMPK直接參與其誘導的腫瘤細胞凋亡,其機製可能與Caspase-3的活性錶達有關.
목적 관찰선감산활화단백격매(AMPK)재동릉초갑소체외유도결장암HT-29세포조망중적작용.방법 농도위1 ～ 50 μmol/L적동릉초갑소분별작용결장암HT-29세포,조단백-DNA매련면역흡부시험(ELISA)법검측동릉초갑소유도적결장암세포조망솔.분광광도법검측작용후적결장암세포반광안선천동안산특이성단백매( Caspase)-3활성,Western blot법측정동릉초갑소작용적종류세포AMPK급기타조망상관단백표체.결과 불동농도동릉초갑소작용우결장암세포후,결장암세포발생조망.수착약물농도증가,세포조망솔야축점증가(P＜0.05),차결장암세포Caspase-3적활성야축점증가(P＜0.05).수착작용시간증가,종류세포p-AMPKα단백조대축점변심변조.동릉초갑소작용적전염AMPKα소간우RNA(siRNA)적세포조망솔(26.33±5.03)％저우전염착의siRNA적세포조망솔(84.40 ±9.70)％,차이유통계학의의(P＜0.05).결론 동릉초갑소유도결장암HT-29세포조망,활화AMPK직접삼여기유도적종류세포조망,기궤제가능여Caspase-3적활성표체유관.
Objective To investigate the apoptosis of HT-29 cells induced by oridonin and the action mechanism.Methods After administration of 1-50 μmol/L oridonin,the enzyme linked immunosorbent assay (ELISA) was used to investigate the apoptosis rate of HT-29 cells induced by oridonin.The expression levels of C-caspase-3 and Amp activated protein kinase (AMPK) proteins were detected by using Western blotting.The caspase-3 activity was measured by using Spectrophotometric assay.Results Different concentrations of oridonin could induce the apoptosis of HT-29 cells and increase the expression of Caspase-3 in a concentration-dependent manner (P ＜ 0.05).With prolonged time,the expression of pAMPKa protein in HT-cells were gradually increased.The apoptosis rate of oridonin-induced HT-29 cells transfected with AMPK small interfering RNA (siRNA) was (26.33 ± 5.03 ) ％,significantly lower than that in the cells transfected with cramble siRNA [ ( 84.40 ± 9.70 ) ％ ] ( P ＜ 0.05 ).Conclusion Apoptosis of HT-29 cells induced by oridonin may be associated with the activation of AMPK,which was probable related with the activation of Caspase-3.
