Piscidinol A对人肝癌HepG-2细胞增殖、凋亡的影响

目的探讨匹西狄醇A(Piscidinol A)对人肝癌Hep G-2细胞增殖及凋亡的影响。方法取对数生长期经胰酶消化后的Hep G-2细胞,培养24 h后分为药物组及对照组,药物组加入不同浓度Piscidinol A(6.25、12.5、25、50、100 mg/L),对照组加入等体积的含10%胎牛血清RPMI1640培养液,12、24、48 h后采用MTT法检测细胞增殖抑制率,计算Piscidinol A的半数抑制浓度(IC50)值;于荧光显微镜下通过Hoechst 33258染色观察凋亡细胞形态,采用AnnexinⅤ-FITC/PI双标记法及流式细胞仪检测细胞凋亡率,Western blo...

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Published in山东医药 Vol. 57; no. 25; pp. 5 - 7
Main Author 肖雪琴 张敏鸿 刘海 杨建琼
Format Magazine Article
LanguageChinese
Published 赣南医学院第一附属医院,江西赣州,341000%赣南医学院 2017
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Summary:目的探讨匹西狄醇A(Piscidinol A)对人肝癌Hep G-2细胞增殖及凋亡的影响。方法取对数生长期经胰酶消化后的Hep G-2细胞,培养24 h后分为药物组及对照组,药物组加入不同浓度Piscidinol A(6.25、12.5、25、50、100 mg/L),对照组加入等体积的含10%胎牛血清RPMI1640培养液,12、24、48 h后采用MTT法检测细胞增殖抑制率,计算Piscidinol A的半数抑制浓度(IC50)值;于荧光显微镜下通过Hoechst 33258染色观察凋亡细胞形态,采用AnnexinⅤ-FITC/PI双标记法及流式细胞仪检测细胞凋亡率,Western blotting法检测B细胞淋巴瘤/白血病基因伴随蛋白x(Bax)蛋白表达。结果药物组Piscidinol A作用后Hep G-2细胞增殖受到抑制且呈量效和时效关系,24 h时IC50值为24.90 mg/L;细胞凋亡染色显示,药物组有典型的细胞凋亡形态出现。药物组6.25、12.5、25、50、100 mg/L的Piscidinol A作用Hep G-2细胞24 h后,细胞凋亡率分别为11.83%、17.49%、21.72%、34.17%、48.51%,对照组为7.23%,随着药物浓度增大,药物组细胞凋亡率相应升高,与对照组相比,P均〈0.05。药物组6.25、12.5、25、50、100 mg/L的Piscidinol A作用Hep G-2细胞24 h后,细胞凋亡相关蛋白Bax表达量分别为0.29、0.33、0.35、0.39、0.43μg/μL,对照组为0.24μg/μL,药物组随着Piscidinol A浓度升高,Bax蛋白表达增加,与对照组相比,P均〈0.05。结论 Piscidinol A能抑制人肝癌Hep G-2细胞增殖并诱导细胞凋亡。
Bibliography:hepatoma;HepG-2 cells;piscidinol A;cells proliferation;apoptosis;B-cell lymphoma/leukemia-2 associated x
Objective To investigate the effects of piscidinol A on apoptosis and proliferation of human liver cancer cell line Hep G-2 and the mechanism. Methods Hep G-2 cells in the logarithmic phase were cultivated for 24 h and then were divided into the drug group and the control group after pancreatic enzyme digesting. The drug group was treated by different concentrations of piscidinol A( 6. 25,12. 5,25,50,and 100 mg/L) and the control group was treated by the same volume RPMI1640 culture which was contained 10% fetal bovine serum. MTT assay was applied to detect the proliferation inhibitory rate at 12,24,and 48 h. IC50 of piscidinol A was calculated,and the morphology of apoptotic cells was observed with Hoechst 33258 fluorescence staining. Flow cytometry was applied to detect apoptosis rate of Hep G-2 cell by using Annexin Ⅴ-FITC/PI double staining. Western blotting was used to detect the B-cell lymphoma/leukem
ISSN:1002-266X
DOI:10.3969/j.issn.1002-266X.2017.25.002