肿瘤研究与临床
腫瘤研究與臨床
종류연구여림상
CANCER RESEARCH AND CLINIC
2012年
2期
76-79
,共4页
张帅%郭卫%任婷婷%卢新昌%彭显博%娄志远%卢群山
張帥%郭衛%任婷婷%盧新昌%彭顯博%婁誌遠%盧群山
장수%곽위%임정정%로신창%팽현박%루지원%로군산
砷剂%肉瘤,Ewing%细胞迁移分析%肿瘤浸润
砷劑%肉瘤,Ewing%細胞遷移分析%腫瘤浸潤
신제%육류,Ewing%세포천이분석%종류침윤
Arsenicals%Sarcoma,Ewing's%Cell migration assays%Neoplasm invasiveness
目的 研究三氧化二砷在体外对尤文肉瘤细胞系迁移和侵袭能力的抑制作用.方法 通过噻唑蓝(MTT)实验选择对尤文肉瘤细胞系(A-673、RD-ES)无毒性的三氧化二砷浓度(＜2μmol/L)后,Transwell迁移和侵袭实验检测三氧化二砷作用下尤文肉瘤细胞的迁移和侵袭能力.Western blot实验及明胶酶谱实验检测PI3K-AKT通路相关作用蛋白及基质金属蛋白酶-9(MMP-9)的表达变化.结果 迁移和侵袭穿过Transwell基底膜的尤文肉瘤细胞的数量随着三氧化二砷浓度的增加而逐渐降低,经不同浓度三氧化二砷处理过的A-673迁移平均数量分别占未处理组的54.3％、49.0％和17.0％,差异有统计学意义(F=112.78,P＜ 0.01),在侵袭实验中分别为52.7％、32.3％和10.3％,差异有统计学意义( F=183.76,P＜ 0.01);经过不同浓度三氧化二砷处理后的RD-ES细胞迁移平均数量分别占未处理组的46.0％、39.0％和8.0％,差异有统计学意义(F=408.25,P＜0.01),在侵袭实验中分别为58.7％、22.3％和9.0％,差异有统计学意义( F=373.25,P＜ 0.01).PI3K-AKT通路相关蛋白和MMP-9表达水平下降.结论 低浓度三氧化二砷可以通过下调PI3K-AKT信号通路的表达来抑制尤文肉瘤细胞系的转移能力.
目的 研究三氧化二砷在體外對尤文肉瘤細胞繫遷移和侵襲能力的抑製作用.方法 通過噻唑藍(MTT)實驗選擇對尤文肉瘤細胞繫(A-673、RD-ES)無毒性的三氧化二砷濃度(＜2μmol/L)後,Transwell遷移和侵襲實驗檢測三氧化二砷作用下尤文肉瘤細胞的遷移和侵襲能力.Western blot實驗及明膠酶譜實驗檢測PI3K-AKT通路相關作用蛋白及基質金屬蛋白酶-9(MMP-9)的錶達變化.結果 遷移和侵襲穿過Transwell基底膜的尤文肉瘤細胞的數量隨著三氧化二砷濃度的增加而逐漸降低,經不同濃度三氧化二砷處理過的A-673遷移平均數量分彆佔未處理組的54.3％、49.0％和17.0％,差異有統計學意義(F=112.78,P＜ 0.01),在侵襲實驗中分彆為52.7％、32.3％和10.3％,差異有統計學意義( F=183.76,P＜ 0.01);經過不同濃度三氧化二砷處理後的RD-ES細胞遷移平均數量分彆佔未處理組的46.0％、39.0％和8.0％,差異有統計學意義(F=408.25,P＜0.01),在侵襲實驗中分彆為58.7％、22.3％和9.0％,差異有統計學意義( F=373.25,P＜ 0.01).PI3K-AKT通路相關蛋白和MMP-9錶達水平下降.結論 低濃度三氧化二砷可以通過下調PI3K-AKT信號通路的錶達來抑製尤文肉瘤細胞繫的轉移能力.
목적 연구삼양화이신재체외대우문육류세포계천이화침습능력적억제작용.방법 통과새서람(MTT)실험선택대우문육류세포계(A-673、RD-ES)무독성적삼양화이신농도(＜2μmol/L)후,Transwell천이화침습실험검측삼양화이신작용하우문육류세포적천이화침습능력.Western blot실험급명효매보실험검측PI3K-AKT통로상관작용단백급기질금속단백매-9(MMP-9)적표체변화.결과 천이화침습천과Transwell기저막적우문육류세포적수량수착삼양화이신농도적증가이축점강저,경불동농도삼양화이신처리과적A-673천이평균수량분별점미처리조적54.3％、49.0％화17.0％,차이유통계학의의(F=112.78,P＜ 0.01),재침습실험중분별위52.7％、32.3％화10.3％,차이유통계학의의( F=183.76,P＜ 0.01);경과불동농도삼양화이신처리후적RD-ES세포천이평균수량분별점미처리조적46.0％、39.0％화8.0％,차이유통계학의의(F=408.25,P＜0.01),재침습실험중분별위58.7％、22.3％화9.0％,차이유통계학의의( F=373.25,P＜ 0.01).PI3K-AKT통로상관단백화MMP-9표체수평하강.결론 저농도삼양화이신가이통과하조PI3K-AKT신호통로적표체래억제우문육류세포계적전이능력.
Objective To investigate the effect of arsenic trioxide (As2O3) on the metastasis capability of Ewing's sarcoma ceils. Methods MTT assays were performed to choose appropriate concentrations of As2O3 (＜ 2 μmol/L) for the experiments.Migration and invasion assays were performed to assess the effect of As2O3 on the metastasis of Ewing's sarcoma cells. Changes in matrix metalloproteinase (MMP)-9 expressions were detected by gel zymography assay and the phosphoinositide 3-kinase/AKT (PI3K-AKT) pathway was investigated using Western blot. Results The amount of Ewing's sarcoma cells across basal membrane of Transwell in migration and invasion assay decreased gradually with the increase in As2O3 concentration. The average quantities of A-673 across the membrane after treatment by gradual concentrations accounted for 54.3 ％,49.0 ％ and 17.0 ％ of that of untreated group respectively in migration assay (F=112.78,P ＜ 0.01), while 52.7 ％, 32.3 ％ and 10.3 ％ in invasion assay (F =183.76, P ＜ 0.01). Similarly, the percentage of RD-ES was 46.0 ％,39.0 ％ and 8.0 ％ in migration assay (F =408.25,P ＜ 0.01) and 58.7 ％,22.3 ％ and 9.0 ％ in invasion assay (F =373.25, P ＜ 0.01) respectively. The difference had statistics significance.The expression of MMP-9 was suppressed by As2O3 treatment according to gel zymography assay.Western blot assay showed that PI3K-AKT pathway was inhibited and nuclear factor kappa B (NF-κB) was inactivated.Conclusion Low-concentration As2O3 may inhibit metastasis capability of Ewing's sarcoma cells.
