Resveratrol contributes to chemosensitivity of malignant mesothelioma cells with activation of p53.
Resveratrol is a naturally occurring polyphenolic phytoalexin with chemopreventive properties. We previously reported a synergistic anti-proliferative effect of resveratrol and clofarabine against malignant mesothelioma (MM) cells. Here, we further investigated molecular mechanisms involved in the synergistic interaction of these compounds in MM MSTO-211H cells. Resveratrol, in combination with clofarabine, time-dependently induced a strong cytotoxic effect with the nuclear accumulation of phospho-p53 (p-p53) in MSTO-211H cells, but not in normal mesothelial MeT-5A cells. Combination treatment up-regulated the levels of p-p53, cleaved caspase-3, and cleaved PARP proteins. Gene silencing with p53-targeting siRNA attenuated the sensitivity of cells to the combined treatment of two compounds. Analyses of p53 DNA binding assay, p53 reporter gene assay, and RTP-CR toward p53-regulated genes, including Bax, PUMA, Noxa and p21, demonstrated that induced p-p53 is transcriptionally active. These results were further confirmed by the siRNA-mediated knockdown of p53 gene. Combination treatment significantly caused the accumulation of cells at G1 phase with the increases in the sub-G0/G1 peak, DNA ladder, nuclear fragmentation, and caspase-3/7 activity. Taken together, these results demonstrate that resveratrol and clofarabine synergistically elicit apoptotic signal via a p53-dependent pathway, and provide a scientific rationale for clinical evaluation of resveratrol as a promising chemopotentiator in MM.
Soonchunhyung Environmental Health Center for Asbestos, College of Medicine, Soonchunhyang University, Cheonan Hospital, Cheonan 330-930, Republic of Korea; Division of Molecular Cancer Research, Soonchunhyang Medical Research Institute, Soonchunhyang University, Cheonan 330-930, Republic of Korea.
Yoon-Jin
YJ
Lee
Department of Pediatrics, Soonchunhyang University Cheonan Hospital, Cheonan 330-721, Republic of Korea.
Ihl-Sung
IS
Park
Soonchunhyung Environmental Health Center for Asbestos, College of Medicine, Soonchunhyang University, Cheonan Hospital, Cheonan 330-930, Republic of Korea.
Yong-Jin
YJ
Lee
Department of Biochemistry, College of Medicine, Soonchunhyang University, Cheonan 330-930, Republic of Korea.
Jung-Hyun
JH
Shim
Division of Molecular Cancer Research, Soonchunhyang Medical Research Institute, Soonchunhyang University, Cheonan 330-930, Republic of Korea.
Moon-Kyun
MK
Cho
Division of Molecular Cancer Research, Soonchunhyang Medical Research Institute, Soonchunhyang University, Cheonan 330-930, Republic of Korea.
Hae-Seon
HS
Nam
Division of Molecular Cancer Research, Soonchunhyang Medical Research Institute, Soonchunhyang University, Cheonan 330-930, Republic of Korea.
Ji Woong
JW
Park
Department of Pediatrics, Soonchunhyang University Cheonan Hospital, Cheonan 330-721, Republic of Korea.
Myung-Ho
MH
Oh
Soonchunhyung Environmental Health Center for Asbestos, College of Medicine, Soonchunhyang University, Cheonan Hospital, Cheonan 330-930, Republic of Korea; Department of Pediatrics, Soonchunhyang University Cheonan Hospital, Cheonan 330-721, Republic of Korea. Electronic address: m1037624@sch.ac.kr.
Sang-Han
SH
Lee
D000227:Adenine Nucleotides / Q000494:pharmacology; D016588:Anticarcinogenic Agents / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000494:pharmacology*; D001087:Arabinonucleosides / Q000494:pharmacology; D001483:Base Sequence; D045744:Cell Line, Tumor; D000077866:Clofarabine; D017931:DNA Primers; D016193:G1 Phase; D016158:Genes, p53; D006801:Humans; D008654:Mesothelioma / Q000188:drug therapy / Q000473:pathology*; D015513:Oncogene Proteins / Q000235:genetics; D016133:Polymerase Chain Reaction; D000077185:Resveratrol; D013267:Stilbenes / Q000494:pharmacology*
D000227:Adenine Nucleotides; D016588:Anticarcinogenic Agents; D000970:Antineoplastic Agents; D001087:Arabinonucleosides; D017931:DNA Primers; D015513:Oncogene Proteins; D013267:Stilbenes; D000077866:Clofarabine; D000077185:Resveratrol
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolum bromide; 4′,6-diamidino-2-phenylindole; Apoptosis; Chemopotentiator; Clofarabine; DAPI; ECL; HRP; MM; MTT; Mesothelioma; RT-PCR; Resveratrol; Reverse transcription-polymerase chain reaction; enhanced chemiluminescence; horseradish peroxidase; malignant mesothelioma; p-p53; p53; phospho-p53