Limits...
High-dose chemotherapeutics of intravesical chemotherapy rapidly induce mitochondrial dysfunction in bladder cancer-derived spheroids.

Yoshida T, Okuyama H, Nakayama M, Endo H, Nonomura N, Nishimura K, Inoue M - Cancer Sci. (2014)

Bottom Line: Two hours after exposure to MMC, the mitochondrial membrane potential decreased and the mitochondria were fragmented, while plasma membrane integrity was maintained.ATP levels rapidly decreased in CTOS after exposure to EPI or MMC.In the CTOS prepared directly from 19 surgical specimens exposed to EPI and MMC, the decrease of ATP levels varied among patients.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Osaka Medical Center for Cancer and Cardiovascular Diseases, Osaka, Japan; Department of Urology, Osaka University Graduate School of Medicine, Osaka, Japan.

Show MeSH

Related in: MedlinePlus

Delivery of chemotherapeutic agents into cancer-tissue originated spheroids (CTOS), and plasma membrane integrity of CTOS after exposure to high-dose chemotherapeutics. (a) Intrinsic fluorescence of epirubicin (EPI) (red) was captured on formalin-fixed, paraffin-embedded CTOS sections. CTOS were exposed to EPI (1 mg/mL) for the indicated time. Nuclei were counterstained with DAPI (blue). Scale bar, 100 μm. (b) Flow cytometric analysis of the intrinsic fluorescence of EPI in CTOS. CTOS were exposed to EPI (1 mg/mL) for the indicated time. (c, d) Loss of plasma membrane integrity was evaluated by propidium iodide (PI) (red) using fluorescence microscopy (c [scale bar, 100 μm]) and flow cytometry (d). CTOS were exposed to control medium or mitomycin C (MMC) (1 mg/mL) for 2 h, and were washed and incubated for an additional 48 h.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4317779&req=5

fig02: Delivery of chemotherapeutic agents into cancer-tissue originated spheroids (CTOS), and plasma membrane integrity of CTOS after exposure to high-dose chemotherapeutics. (a) Intrinsic fluorescence of epirubicin (EPI) (red) was captured on formalin-fixed, paraffin-embedded CTOS sections. CTOS were exposed to EPI (1 mg/mL) for the indicated time. Nuclei were counterstained with DAPI (blue). Scale bar, 100 μm. (b) Flow cytometric analysis of the intrinsic fluorescence of EPI in CTOS. CTOS were exposed to EPI (1 mg/mL) for the indicated time. (c, d) Loss of plasma membrane integrity was evaluated by propidium iodide (PI) (red) using fluorescence microscopy (c [scale bar, 100 μm]) and flow cytometry (d). CTOS were exposed to control medium or mitomycin C (MMC) (1 mg/mL) for 2 h, and were washed and incubated for an additional 48 h.

Mentions: We studied the responses of CTOS to high-dose chemotherapeutics, which are used in IVC for patients with NMIBC after TUR. To provide a reproducible setting, we first analyzed CTOS from a patient-derived xenograft of bladder cancer, BC23.24 We investigated delivery of EPI into CTOS after exposure to high-dose EPI (1 mg/mL), which is used in clinical settings. It is clinically recommended that high-dose chemotherapeutics be retained for 1–2 h after they are instilled into the patient's bladder. Thus, we conducted a 2-h time-course experiment. We took advantage of the intrinsic fluorescence of EPI to monitor drug delivery. Fluorescence microscopy analysis (Fig.2a) and flow cytometric analysis (Fig.2b) revealed that EPI was promptly and homogeneously distributed into the cancer cells comprising CTOS. According to flow cytometric analysis, EPI levels in cancer cells reached a plateau within 1 h. Notably, spheroidal shape was maintained intact even after 2 h exposure to high-dose EPI.


High-dose chemotherapeutics of intravesical chemotherapy rapidly induce mitochondrial dysfunction in bladder cancer-derived spheroids.

Yoshida T, Okuyama H, Nakayama M, Endo H, Nonomura N, Nishimura K, Inoue M - Cancer Sci. (2014)

Delivery of chemotherapeutic agents into cancer-tissue originated spheroids (CTOS), and plasma membrane integrity of CTOS after exposure to high-dose chemotherapeutics. (a) Intrinsic fluorescence of epirubicin (EPI) (red) was captured on formalin-fixed, paraffin-embedded CTOS sections. CTOS were exposed to EPI (1 mg/mL) for the indicated time. Nuclei were counterstained with DAPI (blue). Scale bar, 100 μm. (b) Flow cytometric analysis of the intrinsic fluorescence of EPI in CTOS. CTOS were exposed to EPI (1 mg/mL) for the indicated time. (c, d) Loss of plasma membrane integrity was evaluated by propidium iodide (PI) (red) using fluorescence microscopy (c [scale bar, 100 μm]) and flow cytometry (d). CTOS were exposed to control medium or mitomycin C (MMC) (1 mg/mL) for 2 h, and were washed and incubated for an additional 48 h.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4317779&req=5

fig02: Delivery of chemotherapeutic agents into cancer-tissue originated spheroids (CTOS), and plasma membrane integrity of CTOS after exposure to high-dose chemotherapeutics. (a) Intrinsic fluorescence of epirubicin (EPI) (red) was captured on formalin-fixed, paraffin-embedded CTOS sections. CTOS were exposed to EPI (1 mg/mL) for the indicated time. Nuclei were counterstained with DAPI (blue). Scale bar, 100 μm. (b) Flow cytometric analysis of the intrinsic fluorescence of EPI in CTOS. CTOS were exposed to EPI (1 mg/mL) for the indicated time. (c, d) Loss of plasma membrane integrity was evaluated by propidium iodide (PI) (red) using fluorescence microscopy (c [scale bar, 100 μm]) and flow cytometry (d). CTOS were exposed to control medium or mitomycin C (MMC) (1 mg/mL) for 2 h, and were washed and incubated for an additional 48 h.
Mentions: We studied the responses of CTOS to high-dose chemotherapeutics, which are used in IVC for patients with NMIBC after TUR. To provide a reproducible setting, we first analyzed CTOS from a patient-derived xenograft of bladder cancer, BC23.24 We investigated delivery of EPI into CTOS after exposure to high-dose EPI (1 mg/mL), which is used in clinical settings. It is clinically recommended that high-dose chemotherapeutics be retained for 1–2 h after they are instilled into the patient's bladder. Thus, we conducted a 2-h time-course experiment. We took advantage of the intrinsic fluorescence of EPI to monitor drug delivery. Fluorescence microscopy analysis (Fig.2a) and flow cytometric analysis (Fig.2b) revealed that EPI was promptly and homogeneously distributed into the cancer cells comprising CTOS. According to flow cytometric analysis, EPI levels in cancer cells reached a plateau within 1 h. Notably, spheroidal shape was maintained intact even after 2 h exposure to high-dose EPI.

Bottom Line: Two hours after exposure to MMC, the mitochondrial membrane potential decreased and the mitochondria were fragmented, while plasma membrane integrity was maintained.ATP levels rapidly decreased in CTOS after exposure to EPI or MMC.In the CTOS prepared directly from 19 surgical specimens exposed to EPI and MMC, the decrease of ATP levels varied among patients.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Osaka Medical Center for Cancer and Cardiovascular Diseases, Osaka, Japan; Department of Urology, Osaka University Graduate School of Medicine, Osaka, Japan.

Show MeSH
Related in: MedlinePlus