摘要
目的:研究中药黄芩苷(baicalin)对人Burkitt淋巴瘤细胞株CA46细胞增殖、凋亡的影响并探讨其可能作用机制。方法:应用MTT法绘制细胞生长曲线观察黄芩苷对CA46细胞增殖的影响;Annexin V-FITC/PI双染流式细胞术、细胞DNA片段化、TdT酶介导的原位缺口末端标记(TUNEL)法检测黄芩苷诱导CA46细胞凋亡的能力;RT-PCR法检测黄芩苷作用前后c-myc、bcl-2mRNA表达水平的变化,Western blotting法检测c-Myc、Bcl-2、procaspase-3(caspase-3前体)、PARP(多聚ADP核糖聚合酶)蛋白水平的变化。结果:细胞生长曲线结果显示黄芩苷能明显抑制CA46细胞增殖,半数抑制浓度(IC50)约为10μmol/L;Annexin V-FITC/PI双染流式细胞术早期凋亡的检出、TUNEL晚期凋亡细胞的检出和细胞DNA片段化凋亡梯带的检出,均证实黄芩苷能有效诱导CA46细胞凋亡,细胞凋亡率呈现浓度依赖性递增。黄芩苷作用后CA46细胞c-myc、bcl-2mRNA和c-Myc、Bcl-2、procaspase-3、PARP(116kD)蛋白的表达呈现时间依赖性递减,而PARP(85kD)表达呈现时间依赖性递增。结论:黄芩苷能有效抑制CA46细胞增殖,诱导其凋亡;c-Myc、Bcl-2表达水平下调和caspase-3激活可能参与了这一作用过程。
AIM ： To investigate the effects of baicalin on proliferation inhibition and apoptosis induction in human Burkitt lymphoma cell line CA46 and to explore its underlying mechanisms. METHODS： CA46 cells were exposed to baicalin at different dosages and its proliferation inhibition was detected by MTF assay. The ability of baicalin to induce CA46 cell apoptosis was examined by Annexin V - FITC/PI double staining analysis, TUNEL labeling method and DNA fragmentation. The mRNA expressions of c- myc and bcl -2 were detected by RT- PCR, and the protein expressions of c- Myc, Bcl-2, caspase- 3 precursor （procaspase- 3） and poly ADP- ribese polymerase （PARP） were detected by Western blotting. RFSULTS ： Baicalin remarkably inhibited the CA46 cell proliferation, with an IC50 value of 10μmol/L. Apoptosis was remarkably induced by baicalin in a dose - dependent manner, and its earlier and later stages were detected by annexin V - FITC/PI double staining analysis, TUNEL labeling method and DNA fragmentation, respectively. Furthermore, RT - PCR showed that the mRNA expressions of c - myc and bcl - 2 in treated CA46 cells decreased in a time - dependent manner. Western blotting showed that the protein expressions of c - Myc, Bcl - 2, procaspase - 3 and PARP （116 kD） in baicalin treated CA46 cells were down - regulated in a time - dependent manner, while the expression of PARP（85 kD） was upregulated. CONCLUSION： Baicalin efficiently induces proliferation inhibition and apoptosis in CA46 cells, which may be related with the down - regulation of c - Myc and Bcl - 2 expressions, as well as the up - regulation of caspase - 3 activity.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2009年第5期888-892,共5页
Chinese Journal of Pathophysiology
基金
福建省高校新世纪优秀人才计划资助项目(No.NCETFJ-0604)
福建医科大学教授基金资助项目(No.JS06081)
