Hematoporphyrin monomethyl ether combined with He-Ne laser irradiation-induced apoptosis in canine breast cancer cells through the mitochondrial pathway.
10.4142/jvs.2016.17.2.235
- Author:
Huatao LI
1
;
Jinjin TONG
;
Jun BAO
;
Damu TANG
;
Wenru TIAN
;
Yun LIU
Author Information
1. Department of Clinic Veterinary Medicine, College of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China. abliuyun@yeah.net
- Publication Type:Original Article
- Keywords:
CHMm cells;
apoptosis;
hematoporphyrin monomethyl ether;
mitochondrial pathway;
promising photodynamic therapy
- MeSH:
Apoptosis*;
Blotting, Western;
Breast Neoplasms*;
Breast*;
Caspase 3;
Caspase 9;
Cell Death;
Cytochromes c;
Cytoplasm;
DNA;
DNA Fragmentation;
DNA Nucleotidylexotransferase;
Ether*;
Fluorescein;
Genes, p53;
Hematoporphyrins*;
In Vitro Techniques;
Mitochondria;
Photochemotherapy;
RNA, Messenger;
Vacuoles
- From:Journal of Veterinary Science
2016;17(2):235-242
- CountryRepublic of Korea
- Language:English
-
Abstract:
Hematoporphyrin monomethyl ether (HMME) combined with He-Ne laser irradiation is a novel and promising photodynamic therapy (PDT)-induced apoptosis that can be applied in vitro on canine breast cancer cells. However, the exact pathway responsible for HMME-PDT in canine breast cancer cells remains unknown. CHMm cells morphology and apoptosis were analyzed using optical microscope, terminal deoxynucleotidyl transferase dUTP nick end labeling fluorescein staining and DNA ladder assays. Apoptotic pathway was further confirmed by Real-time-polymerase chain reaction and Western blotting assays. Our results showed that HMME-PDT induced significant changes in cell morphology, such as formation of cytoplasmic vacuoles and the gradual rounding of cells coupled with decreased size and detachment. DNA fragmentation and cell death was shown to occur in a time-dependent manner. Furthermore, HMME-PDT increased the activities of caspase-9 and caspase-3, and released cytochrome c from mitochondria into the cytoplasm. HMME-PDT also significantly increased both mRNA and protein levels of Bax and decreased P53 gene expression in a time-dependent manner, while the mRNA and protein expression of Bcl-2 were repressed. These alterations suggest that HMME-PDT induced CHMm cell apoptosis via the mitochondrial apoptosis pathway and had anti-canine breast cancer effects in vitro.
