Institute of Atomic and Molecular Sciences, Academia Sinica, Taipei 10617, Taiwan.
Int J Mol Sci. 2022 Dec 13;23(24):15799. doi: 10.3390/ijms232415799.
Time-gated fluorescence lifetime imaging microscopy with the -BMVC fluorescent probe provides a visualizing method for the study of exogenous G-quadruplexes (G4s) in live cancer cells. Previously, imaging results showed that the parallel G4s are accumulated and that nonparallel G4s are not detected in the lysosomes of CL1-0 live cells. In this work, the detection of the G4 signals from exogenous GTERT-d(FN) G4s in the lysosomes may involve a structural change in live cells from intramolecular nonparallel G4s to intermolecular parallel G4s. Moreover, the detection of the G4 signals in the lysosomes after the 48 h incubation of HT23 G4s with CL1-0 live cells indicates the occurrence of structural conversion from the nonparallel G4s to the parallel G4s of HT23 in the live cells. In addition, the detection of much stronger G4 signals from ss-GTERT-d(FN) than ss-HT23 in the lysosomes of CL1-0 live cells may be explained by the quick formation of the intermolecular parallel G4s of ss-GTERT-d(FN) and the degradation of ss-HT23 before its intramolecular parallel G4 formation. This work provides a new approach to studying G4-lysosome interactions in live cells.
基于 -BMVC 荧光探针的时间门控荧光寿命成像显微镜为研究活细胞中外源性 G-四链体 (G4s) 提供了一种可视化方法。以前的成像结果表明，平行 G4s 在 CL1-0 活细胞的溶酶体中积累，而非平行 G4s 则无法检测到。在这项工作中，溶酶体中来自外源性 GTERT-d(FN) G4s 的 G4 信号的检测可能涉及活细胞中从分子内非平行 G4s 到分子间平行 G4s 的结构变化。此外，在 HT23 G4s 与 CL1-0 活细胞孵育 48 小时后，在溶酶体中检测到 G4 信号表明 HT23 中的非平行 G4s 在活细胞中发生了结构转换为平行 G4s。此外，在 CL1-0 活细胞的溶酶体中，ss-GTERT-d(FN) 的 G4 信号比 ss-HT23 强得多，这可能是由于 ss-GTERT-d(FN) 快速形成分子间平行 G4s 以及 ss-HT23 在形成分子内平行 G4s 之前降解所致。这项工作为研究活细胞中的 G4-溶酶体相互作用提供了一种新方法。
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