BACKGROUND AND OBJECTIVERadiotherapy (RT) is a major non-surgical modality in the comprehensive treatment for colorectal adenocarcinoma. The radioresistance of cancer stem cells (CSCs) is a key factor that influences therapeutic effectiveness. This study was to investigate the effects of specific chromosome structure and histone modification in CSCs in colorectal adenocarcinoma radioresistance.

METHODSSamples were collected from resected human colorectal adenocarcinomas. Subcutaneous colorectal cancer model was established in nude mice. Immunohistochemistry showed that xenografts generated from bulk colorectal cancer cells resembled the original tumor specimen. Flow cytometry was performed to sort CSCs (CD133+) and non-CSCs (CD133-) from both resected samples of colorectal adenocarcinoma and xenograft before and after high single-dose radiation. The markers labeling heterochromatin (H3K9me3, HP1-alpha and H3K4me1) and euchromatin (H3K4me3) in CD133+ and CD133- nucleus were detected by immunofluorescence.

RESULTSThere was distinct difference in chromatin structure between colorectal CSCs (CD133+) and non-CSCs (CD133-). The chromatin displayed compact patches in CD133+ nucleus, but loosely latticed structure in CD133- nucleus; immunofluorescence verified that the compact patches existing in CSCs was generated from heterochromatin construction. In addition, the vacuole-like defect in heterochromatin regions of CSCs was observed within 24 h after exposure to 10 gray (Gy) single-dose RT. Interestingly, this phenomenon was repaired from 96 h, and recovered to dense plaque structure in heterochromatin regions of CSCs after 144 h. However, no significant difference in non-CSCs was observed after RT exception for a loose chromatin structure.

CONCLUSIONSCSCs play a role in radiosensitivity in colorectal cancer. The mechanism may be related to heterochromatin formation and histone methylation.

AC133 Antigen ; Adenocarcinoma ; pathology ; radiotherapy ; Adult ; Aged ; Aged, 80 and over ; Animals ; Antigens, CD ; metabolism ; Cell Nucleus ; genetics ; pathology ; Colorectal Neoplasms ; pathology ; radiotherapy ; Female ; Glycoproteins ; metabolism ; Heterochromatin ; metabolism ; Histones ; metabolism ; Humans ; Male ; Methylation ; Mice ; Mice, Nude ; Middle Aged ; Neoplasm Transplantation ; Neoplastic Stem Cells ; pathology ; radiation effects ; Particle Accelerators ; Peptides ; metabolism ; Radiation Dosage ; Radiation Tolerance

14-3-3zeta is related to many cancer survival cellular processes. In a previous study, we showed that silencing 14-3-3zeta decreases the resistance of hepatocellular carcinoma (HCC) to chemotherapy. In this study, we investigated whether silencing 14-3-3zeta affects the radioresistance of cancer stem-like cells (CSCs) in HCC. Knockdown of 14-3-3zeta decreased cell viability and the number of spheres by reducing radioresistance in CSCs after gamma-irradiation (IR). Furthermore, the levels of pro-apoptotic proteins were upregulated in CSCs via silencing 14-3-3zeta after IR. These results suggest that 14-3-3zeta knockdown enhances radio-induced apoptosis by reducing radioresistance in liver CSCs.
14-3-3 Proteins/genetics/*metabolism ; Antigens, CD/genetics/*metabolism ; Apoptosis Regulatory Proteins/genetics/metabolism ; Carcinoma, Hepatocellular/genetics/metabolism ; Cell Line, Tumor ; *Gamma Rays ; Glycoproteins/genetics/*metabolism ; Humans ; Liver Neoplasms/genetics/metabolism ; Neoplastic Stem Cells/metabolism/*radiation effects ; Peptides/genetics/*metabolism ; *Radiation Tolerance