Visualizing cell-cycle kinetics after hypoxia/reoxygenation in HeLa cells expressing fluorescent ubiquitination-based cell cycle indicator (Fucci)

Goto, Tatsuaki; Kaida, Atsushi; Miura, Masahiko

doi:10.1016/j.yexcr.2015.10.019

Cited by 18 publications

(23 citation statements)

References 18 publications

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“…To further verify the ability of CaM‐PB to mediate the physiological status of the TME, cell cycles were detected using flow cytometry. [ 17 ] Compared with the Ca‐PB (26.2% cells in S phase) and CaM groups (28.3% cells in S phase), 51.2% and 47.4% of cells were found in S phase after treatment with CaM‐PB and SiM‐PB, respectively, suggesting hypoxia relief by MnO 2 and efficient cellular uptake mediated by BSA (Figure 2H; Figure S5, Supporting Information). HCPT is a cell cycle‐specific drug that acts on tumor cells in the S phase, and the improvement of hypoxia that promotes cells into the S phase may significantly amplify the chemotherapeutic effect.…”

Section: Resultsmentioning

confidence: 99%

Smart Tumor Microenvironment‐Responsive Nanotheranostic Agent for Effective Cancer Therapy

Guo

Sun

et al. 2020

Adv Funct Materials

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The tumor microenvironment (TME), which includes acidic and hypoxic conditions, severely impedes the therapeutic efficacy of antitumor agents. Herein, MnO 2 -loaded, bovine serum albumin, and PEG co-modified mesoporous CaSiO 3 nanoparticles (CaM-PB NPs) are developed as a nanoplatform with sequential theranostic functions for the engineering of TME. The MnO 2 NPs generate O 2 in situ by reacting with endogenous H 2 O 2 , relieving the hypoxic state of the TME that further modulates the cancer cell cycle status to S phase, which improves the potency of co-loaded S phase-sensitive chemotherapeutic drugs. After the hypoxia relief, CaM-PB can sustainably release drugs due to the enlarged pores of mesoporous CaSiO 3 in the acidic TME, preventing the drug pre-leakage into the blood circulation and insufficient drug accumulation at tumor sites. Moreover, the Mn 2+ released from the MnO 2 NPs at tumor sites can potentially serve as a diagnostic agent, enabling the identification of tumor regions by T 1 -weighted magnetic resonance imaging during therapy. In vivo pharmacodynamics results demonstrate that these synergetic effects caused by CaM-PB NPs significantly contribute to the inhibition of tumor progression. Therefore, the CaM-PB NPs with sequential theranostic functions are a promising system for effective cancer therapy. Keywordshypoxia relief, mesoporous calcium silicate, sequential functions, sustained drug release, synergetic effects

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Section: Resultsmentioning

confidence: 99%

Smart Tumor Microenvironment‐Responsive Nanotheranostic Agent for Effective Cancer Therapy

Guo

Sun

et al. 2020

Adv Funct Materials

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“…If G2 arrest was determined only by DSB-associated events, then the elongation of G2 arrest could be explained by the fact that DSB repair is slower after irradiation under hypoxia. Furthermore, DSB generation due to reoxygenation after hypoxia might also be involved [ 14 , 26 ]. However, our observation that G2 arrest is enhanced when cells were irradiated in red phase could not be explained by the notion proposed by Karanam et al [ 23 ], as described above.…”

Section: Discussionmentioning

confidence: 99%

“…Cell sorting and flow-cytometric analysis was performed as described previously [ 14 ]. Briefly, exponentially growing HeLa-Fucci cells were sorted on a MoFlo XDP (Beckman Coulter, Brea, CA, USA) according to their Fucci fluorescence.…”

Section: Methodsmentioning

confidence: 99%

Temporo-spatial cell-cycle kinetics in HeLa cells irradiated by Ir-192 high dose-rate remote afterloading system (HDR-RALS)

et al. 2016

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BackgroundIntracavitary irradiation plays a pivotal role in definitive radiotherapy for cervical cancer, and the Ir-192 high dose-rate remote afterloading system (HDR-RALS) is often used for this purpose. Under this condition, tumor tissues receive remarkably different absorption doses, with a steep gradient, depending on distance from the radiation source. To obtain temporo-spatial information regarding cell-cycle kinetics in cervical cancer following irradiation by Ir-192 HDR-RALS, we examined HeLa cells expressing the fluorescence ubiquitination-based cell cycle indicator (Fucci), which allowed us to visualize cell-cycle progression.MethodsHeLa-Fucci cells, which emit red and green fluorescence in G1 and S/G2/M phases, respectively, were grown on 35-mm dishes and irradiated by Ir-192 HDR-RALS under normoxic and hypoxic conditions. A 6 French (Fr) catheter was used as an applicator. A radiation dose of 6 Gy was prescribed at hypothetical treatment point A, located 20 mm from the radiation source. Changes in Fucci fluorescence after irradiation were visualized for cells from 5 to 20 mm from the Ir-192 source. Several indices, including first green phase duration after irradiation (FGPD), were measured by analysis of time-lapse images.ResultsCells located 5 to 20 mm from the Ir-192 source became green, reflecting arrest in G2, in a similar manner up to 12 h after irradiation; at more distant positions, however, cells were gradually released from the G2 arrest and became red. This could be explained by the observation that the FGPD was longer for cells closer to the radiation source. Detailed observation revealed that FGPD was significantly longer in cells irradiated in the green phase than in the red phase at positions closer to the Ir-192 source. Unexpectedly, the FGPD was significantly longer after irradiation under hypoxia than normoxia, due in large part to the elongation of FGPD in cells irradiated in the red phase.ConclusionUsing HeLa-Fucci cells, we obtained the first temporo-spatial information about cell-cycle kinetics following irradiation by Ir-192 HDR-RALS. Our findings suggest that the potentially surviving hypoxic cells, especially those arising from positions around point A, exhibit different cell-cycle kinetics from normoxic cells destined to be eradicated.Electronic supplementary materialThe online version of this article (doi:10.1186/s13014-016-0669-8) contains supplementary material, which is available to authorized users.

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“…Some very well vascularized large tumors may have a greater fraction of cycling cells. Haass et al [31] and Goto et al [32] demonstrated that FUCCI-expressing cancer cells were arrested in G 1 phase during hypoxia and restarted proliferation when oxygen was supplied. Chittajallu et al [33] also performed intravital FUCCI imaging, in their case, of HT1080 fibrosarcoma cells in a dorsal skin-fold chamber in nude mice and demonstrated that tumors in the chamber comprised approximately 60% of cancer cells in G 1 phase and approximately 29% of cells in S/G 2 /M phases, similar to the results described above.…”

Section: Real-time Fucci Imaging Of Cell-cycle Dynamics Of Cancer Celmentioning

confidence: 99%

FUCCI Real-Time Cell-Cycle Imaging as a Guide for Designing Improved Cancer Therapy: A Review of Innovative Strategies to Target Quiescent Chemo-Resistant Cancer Cells

Yano

Tazawa

Kagawa

et al. 2020

Cancers

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Progress in chemotherapy of solid cancer has been tragically slow due, in large part, to the chemoresistance of quiescent cancer cells in tumors. The fluorescence ubiquitination cell-cycle indicator (FUCCI) was developed in 2008 by Miyawaki et al., which color-codes the phases of the cell cycle in real-time. FUCCI utilizes genes linked to different color fluorescent reporters that are only expressed in specific phases of the cell cycle and can, thereby, image the phases of the cell cycle in real-time. Intravital real-time FUCCI imaging within tumors has demonstrated that an established tumor comprises a majority of quiescent cancer cells and a minor population of cycling cancer cells located at the tumor surface or in proximity to tumor blood vessels. In contrast to most cycling cancer cells, quiescent cancer cells are resistant to cytotoxic chemotherapy, most of which target cells in S/G2/M phases. The quiescent cancer cells can re-enter the cell cycle after surviving treatment, which suggests the reason why most cytotoxic chemotherapy is often ineffective for solid cancers. Thus, quiescent cancer cells are a major impediment to effective cancer therapy. FUCCI imaging can be used to effectively target quiescent cancer cells within tumors. For example, we review how FUCCI imaging can help to identify cell-cycle-specific therapeutics that comprise decoy of quiescent cancer cells from G1 phase to cycling phases, trapping the cancer cells in S/G2 phase where cancer cells are mostly sensitive to cytotoxic chemotherapy and eradicating the cancer cells with cytotoxic chemotherapy most active against S/G2 phase cells. FUCCI can readily image cell-cycle dynamics at the single cell level in real-time in vitro and in vivo. Therefore, visualizing cell cycle dynamics within tumors with FUCCI can provide a guide for many strategies to improve cell-cycle targeting therapy for solid cancers.

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Visualizing cell-cycle kinetics after hypoxia/reoxygenation in HeLa cells expressing fluorescent ubiquitination-based cell cycle indicator (Fucci)

Cited by 18 publications

References 18 publications

Smart Tumor Microenvironment‐Responsive Nanotheranostic Agent for Effective Cancer Therapy

Smart Tumor Microenvironment‐Responsive Nanotheranostic Agent for Effective Cancer Therapy

Temporo-spatial cell-cycle kinetics in HeLa cells irradiated by Ir-192 high dose-rate remote afterloading system (HDR-RALS)

FUCCI Real-Time Cell-Cycle Imaging as a Guide for Designing Improved Cancer Therapy: A Review of Innovative Strategies to Target Quiescent Chemo-Resistant Cancer Cells

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