中国比较医学杂志
中國比較醫學雜誌
중국비교의학잡지
CHINESE JOURNAL OF COMPARATIVE MEDICINE
2014年
10期
7-11,17
,共6页
翟斡%沈一平%沈嫣婧%楼正青%丁志山
翟斡%瀋一平%瀋嫣婧%樓正青%丁誌山
적알%침일평%침언청%루정청%정지산
小豆蔻明%K562细胞%凋亡%PTEN%p-Akt%NF-κB%Bcl-2
小豆蔻明%K562細胞%凋亡%PTEN%p-Akt%NF-κB%Bcl-2
소두구명%K562세포%조망%PTEN%p-Akt%NF-κB%Bcl-2
Cardamonin%K562 cells%Apoptosis,PTEN,p-Akt,NF-κB,Bcl-2
目的:探讨小豆蔻明诱导K562细胞凋亡与PTEN、p-Akt、NF-κB、Bcl-2等抑凋亡蛋白及促凋亡蛋白表达量的关系。方法①普通光镜观察小豆蔻明作用K562细胞48 h后的细胞形态学变化。②采用MTT法检测小豆蔻明作用K562细胞48 h后的凋亡情况,计算IC50。③应用流式细胞仪检测小豆蔻明作用K562细胞48 h后的细胞凋亡率。④采用RT-PCR技术检测小豆蔻明作用K562细胞48 h后Bcl-2、Bax的mRNA表达量。⑤采用Western blot技术检测小豆蔻明作用K562细胞48 h后PTEN、p-Akt、NF-κB、Bcl-2的蛋白表达量。结果小豆蔻明作用K562细胞48 h后,形态学出现明显凋亡现象。 MTT提示K562细胞增殖抑制明显且呈量效和时效关系。早期凋亡率与空白组比有显著增加(P<0.05),呈剂量依赖关系。 Bcl-2 mRNA的表达较空白组明显下降(P<0.05), Bax mRNA的表达较空白组明显升高(P<0.05),呈现浓度依赖性。 PTEN蛋白表达量随小豆蔻明的浓度增加明显增加,而p-Akt、NF-κB、Bcl-2蛋白表达量则减少。结论小豆蔻明通过增加PTEN蛋白的表达量,同时下调p-Akt、NF-κB、Bcl-2蛋白的表达量促进K562细胞的凋亡,且呈剂量依赖性。
目的:探討小豆蔻明誘導K562細胞凋亡與PTEN、p-Akt、NF-κB、Bcl-2等抑凋亡蛋白及促凋亡蛋白錶達量的關繫。方法①普通光鏡觀察小豆蔻明作用K562細胞48 h後的細胞形態學變化。②採用MTT法檢測小豆蔻明作用K562細胞48 h後的凋亡情況,計算IC50。③應用流式細胞儀檢測小豆蔻明作用K562細胞48 h後的細胞凋亡率。④採用RT-PCR技術檢測小豆蔻明作用K562細胞48 h後Bcl-2、Bax的mRNA錶達量。⑤採用Western blot技術檢測小豆蔻明作用K562細胞48 h後PTEN、p-Akt、NF-κB、Bcl-2的蛋白錶達量。結果小豆蔻明作用K562細胞48 h後,形態學齣現明顯凋亡現象。 MTT提示K562細胞增殖抑製明顯且呈量效和時效關繫。早期凋亡率與空白組比有顯著增加(P<0.05),呈劑量依賴關繫。 Bcl-2 mRNA的錶達較空白組明顯下降(P<0.05), Bax mRNA的錶達較空白組明顯升高(P<0.05),呈現濃度依賴性。 PTEN蛋白錶達量隨小豆蔻明的濃度增加明顯增加,而p-Akt、NF-κB、Bcl-2蛋白錶達量則減少。結論小豆蔻明通過增加PTEN蛋白的錶達量,同時下調p-Akt、NF-κB、Bcl-2蛋白的錶達量促進K562細胞的凋亡,且呈劑量依賴性。
목적:탐토소두구명유도K562세포조망여PTEN、p-Akt、NF-κB、Bcl-2등억조망단백급촉조망단백표체량적관계。방법①보통광경관찰소두구명작용K562세포48 h후적세포형태학변화。②채용MTT법검측소두구명작용K562세포48 h후적조망정황,계산IC50。③응용류식세포의검측소두구명작용K562세포48 h후적세포조망솔。④채용RT-PCR기술검측소두구명작용K562세포48 h후Bcl-2、Bax적mRNA표체량。⑤채용Western blot기술검측소두구명작용K562세포48 h후PTEN、p-Akt、NF-κB、Bcl-2적단백표체량。결과소두구명작용K562세포48 h후,형태학출현명현조망현상。 MTT제시K562세포증식억제명현차정량효화시효관계。조기조망솔여공백조비유현저증가(P<0.05),정제량의뢰관계。 Bcl-2 mRNA적표체교공백조명현하강(P<0.05), Bax mRNA적표체교공백조명현승고(P<0.05),정현농도의뢰성。 PTEN단백표체량수소두구명적농도증가명현증가,이p-Akt、NF-κB、Bcl-2단백표체량칙감소。결론소두구명통과증가PTEN단백적표체량,동시하조p-Akt、NF-κB、Bcl-2단백적표체량촉진K562세포적조망,차정제량의뢰성。
ObjectiveTo observe the apoptotic effect of cardamonin on K562 cells and its relationship with the expressions of PTEN, p-Akt, NF-κB and Bcl-2.Methods K562 cells were treated with cardamonin for 48 h, and the following tests were performed:(1) The cell morphology was observed by light microscopy.(2)IC50 of the K562 cells was dtermined by MTT test.(3) The apoptosis rate was detected by flow cytometry.(4) The expressions of Bcl-2 and Bax mRNA were detected a by RT-PCR.(5) The expressions of PTEN, p-Akt, NF-κB and Bcl-2 proteins were detected by Western blot.Results Obvious apoptosis was observed in the K562 cells after treated with cardamonin for 48 h.MTT assay indicated that the proliferation of K562 cells was obviously inhibited in a dose-and time-dependent manner. Comparing with the blank group, the early apoptosis rate and expression of Bax mRNA were significantly increased.At the same time, the expression of Bcl-2 mRNA was significantly decreased.All of them presented a dose-dependent manner. The expression of PTEN obviously increased with the increasing dose of cardamonin and the expressions of p-Akt, NF-κB and Bcl-2 were decreased.Conclusions Cardamonin promotes the apoptosis in K562 cells in a dose-dependent manner by increasing the expression of PTEN and decreasing the expressions of p-Akt, NF-κB, and Bcl-2.