流式细胞术联合共聚焦显微镜对G_2和M期作用位点的鉴别意义

Discrimination of Anticancer Agent Action Loci at G_2 and M phases by Flow Cytometry and Confocal Microscopic Imaging

本研究探讨抗癌药物在细胞周期G2期和M期作用位点的鉴别方法。用G2期和M期细胞阻滞剂的甲安吖啶(m-AMSA)和长春花碱(VBL),分别诱导MOLT-4细胞产生细胞G2期和M期阻滞,用流式细胞术分析DNA直方图含量变化,共聚焦显微镜观察阻滞后细胞形态,并寻找上述两种药物诱导阻滞成功后的差异。结果表明:流式细胞术有检测显示,诱导阻滞成功的MOLT-4细胞均表现为G2/M峰增高,但仅凭流式细胞术单独检测无法判别两者差异。共聚焦显微镜观察显示甲安吖啶诱导G2期阻滞的细胞呈现G2期形态特征,VBL诱导M期阻滞的细胞呈现M期形态特征,共聚焦显微镜对细胞形态学的观察有助于区分两者的差异。因此,在流式细胞术验证诱导成功的基础上进一步经共聚焦显微镜观察有助于鉴别G2和M期的阻滞剂。结论:流式细胞术与共聚焦显微镜技术结合是判别抗癌药物的G期和M期细胞阻滞剂类型的有效方法之一。

This study was purposed to evaluate a method to discriminate the action loci of anticancer agents in G2 and M phases of cell cycle. The methamsacrine （m-AMSA） and vinblastine （VBL） , already known as G2 and M phase arrest agent respectively, were used to induce the arrest of MOLT-4 cells at G2 and M phases, the change of DNA content was detected by flow cytometry, the morphology of arrested cells was observed by confocal microscopy so as to find the arrest efficacy difference of 2 anticancer agents. As a result, the flow cytometric detection showed that the arrested MOLT-4 cells displayed the raise of peaks in G2 and M phases, but flow cytometric detection None can not discriminate the difference between them. The observation with confocal mioroscopy showed that the MOLT-4 cells arrested by m-AMSA displayed the morphologic features in G2 phase, while the MOLT-4 cells arrested by VBL displayed the morphologic features in M phase. This observation with confocal microscopy is helpful to discriminate the difference between them. In conclusion, the combination of flow cytometry with confocN microscopy is one of the effective methods to discriminate the kind of G2 or M phase arresting agent of anticancer drugs.