Chidamide and mitomycin C exert synergistic cytotoxic effects against bladder cancer cells in vitro and suppress tumor growth in a rat bladder cancer model

Wang, Shao-Chuan; Yu, Chih Teng; Wu, Yao‐Cheng; Chang, Ya-Chuan; Chen, Sung‐Lang; Sung, Wen‐Wei

doi:10.1016/j.canlet.2022.01.007

Cited by 19 publications

(22 citation statements)

References 24 publications

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“…The MTT reagent (0.5 mg/mL) was added to the wells and incubated for 3 h at 37 • C, followed by resolution in dimethyl sulfoxide. The absorbance (optical density) was measured at 570 nm with an ELISA plate reader [25,26].…”

Section: Cell Viability Assaymentioning

confidence: 99%

“…Cell cycle analysis and apoptosis analysis were performed using a FACSCanto TM II Cell Analyzer (BD Biosciences, Franklin Lakes, NJ, USA) [25,26]. The cell lines were first cultured in a six-well plate and treated with PNU-74654 at doses of 0, 50, and 150 µM for 72 h. For analysis of the cell cycle distribution, the harvested cells were fixed with cold 70% ethanol overnight, resuspended in phosphate buffered saline (PBS) containing 4 µg/mL propidium iodide (PI) and 0.5 mg/mL RNase A, and analyzed by flow cytometry.…”

Section: Flow Cytometrymentioning

confidence: 99%

“…After washing with PBS and staining with 10 µg/mL Hoechst 33,342 (Invitrogen, Carlsbad, CA, USA), apoptotic cells were detected by fluorescence microscopy (ImageXpress PICO, San Jose, CA, USA) at excitation wavelengths of 350-390 nm and emission wavelengths of 420-480 nm. The apoptotic cells were identified by their condensed and fragmented nuclei, and their percentages were calculated in five different fields at 20× magnification [25,26].…”

Section: Hoechst 33342 Stainingmentioning

confidence: 99%

“…A horseradish peroxidase-conjugated antibody was incubated with the membranes, followed by a chemiluminescent reagent, and the cells were examined with an ImageQuant LAS4000 instrument (GE Healthcare, Marlborough, MA, USA). ImageJ software was used to analyze the integrated density of each membrane [25,26].…”

Section: Human Oncology Array For Proteome Profilingmentioning

confidence: 99%

“…The membranes were washed with Tris-buffered saline containing Tween 20 (TBST), incubated with secondary antibodies for 1 h. Afterwards, the membranes were washed again with TBST and treated with Immobilon TM Western Chemiluminescent HRP Substrate (Merck Millipore, Burlington, MA, USA). The proteins were detected with a ImageQuant LAS4000 instrument (GE Healthcare, Marlborough, MA, USA) [25,26].…”

Section: Protein Extraction and Western Blottingmentioning

confidence: 99%

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The Therapeutic Role of PNU-74654 in Hepatocellular Carcinoma May Involve Suppression of NF-κB Signaling

Wang

Chang

et al. 2022

Medicina

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Background and Objectives: PNU-74654, a Wnt/β-catenin inhibitor, has reported antitumor activities; however, the therapeutic potential of PNU-74654 in hepatocellular carcinoma (HCC) has not been investigated in detail. The aim of this study was to clarify the cytotoxic effects of PNU-74654 against HCC and to uncover its molecular mechanism. Materials and Methods: HepG2 and Huh7 liver cancer cell lines were selected to determine the antitumor properties of PNU-74654. Survival of the liver cancer cells in response to PNU-74654 was assessed by cell viability assays, and the apoptosis effect of PNU-74654 was analyzed by flow cytometry and visualized by Hoechst staining. An oncology array was used to explore the underlying molecular routes of PNU-74654 action in the cells. The migration properties were examined with a wound healing assay, and western blotting was conducted to evaluate protein expression. Results: Treatment with PNU-74654 decreased cell viability and inhibited cell migration. The cell cycle analysis and Hoechst staining revealed an increase in the population of cells at the sub-G1 stage and apoptotic morphological changes in the nucleus. The oncology array identified 84 oncology-related proteins and a suppressed expression of Bcl-xL and survivin. Western blotting showed that PNU-74654 could interfere with cell cycle-related proteins through the NF-κB pathway. Conclusions: PNU-74654 shows antiproliferative and antimigration effects against HepG2 and Huh7 cells, and its antitumor activity may be attributable to its interference in cell cycle regulation and the NF-κB pathway.

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Section: Cell Viability Assaymentioning

confidence: 99%

Section: Flow Cytometrymentioning

confidence: 99%

Section: Hoechst 33342 Stainingmentioning

confidence: 99%

Section: Human Oncology Array For Proteome Profilingmentioning

confidence: 99%

Section: Protein Extraction and Western Blottingmentioning

confidence: 99%

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