[关键词]
[摘要]
目的: 探讨人工合成的BH3-HIV-TAT肽对前列腺癌细胞株LNCaP增殖的影响。方法: 以25、50、1 000 μmol/L的BH3-HIV-TAT处理LNCaP细胞12或24 h,以激光共聚焦显微镜观察BH3-HIV-TAT肽的细胞定位;观察经BH3-HIV-TAT肽处理后,LNCap细胞凋亡过程中核染色质的形态学变化;用流式细胞仪分析不同时间不同浓度的短肽对细胞凋亡程度和周期的影响;以MTS/PMS测定,分析短肽对LNCaP细胞的增殖是否有浓度依赖性的影响。结果: 激光共聚焦检测结果显示BH3-HIV-TAT肽主要定位在细胞核内;LNCap细胞经短肽处理后,可观察到分为2期的细胞凋亡过程;流式细胞仪检测结果证实了短肽对细胞凋亡程度和周期的影响;MTS/PMS测定表明该短肽具有浓度依赖性的抗前列腺癌细胞的活性。〖HT5W〗结论〖HT5"SS〗: BH3 HIV-TAT肽可有效地诱导前列腺癌细胞株LNCaP的凋亡,从而影响其增殖,实验结果为癌症靶向性治疗提供了依据。
[Key word]
[Abstract]
Objective: To investigate the influence of BH3-HIV-TAT peptide on proliferation of prostate cancer cell line LNCap.Methods: LNCap cells were treated with BH3-HIV-TAT (25, 50, and 100 μmol/L) for 12 or 24 h. Then, laser scanning confocal fluorescence microscope was used to observe the cellular localization of BH3-HIV-TAT; Hoechst33258 staining was used to observe the morphological changes of LNCap cells during apoptosis; flow cytometry (FCM) was applied to study cell apoptosis and cell cycle of LNCap cells; and MTS/PMS assay was used to analyze whether the effect of BH3-HIV-TAT peptide was concentration dependent.Results: 〖WTBZ〗It was found that BH3-HIV-TAT peptide was mainly located in cell nucleus. The apoptosis of LNCap cells had 2 stages after BH3-HIV-TAT peptide treatment. Flow cytometry confirmed the peptide arrested cell cycle and induced apoptosis of LNCap cells. MTS/PMS assay showed BH3-HIV-TAT peptide had a concentration-dependent anti-LNCap cell effect. Conclusion: BH3-HIV-TAT peptide can effectively induce apoptosis of LNCap cells, influencing the proliferation of the cells, which provide an evidence for targeted therapy of cancer.
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[基金项目]
国家高新技术发展规划(“863”计划)(2001AA215321)