Effects of heterochromatin in colorectal cancer stem cells on radiosensitivity.
OBJECTIVE
Radiotherapy (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.


METHODS
Samples 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.


RESULTS
There 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.


CONCLUSIONS
CSCs play a role in radiosensitivity in colorectal cancer. The mechanism may be related to heterochromatin formation and histone methylation.
Chen
Ting
T
Department of Thoracic Oncology, Cancer Center, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P. R. China.
Zhang
Yu
Y
Guo
Wen-Hao
WH
Meng
Mao-Bin
MB
Mo
Xian-Ming
XM
Lu
You
Y
D000071916:AC133 Antigen; D000230:Adenocarcinoma / Q000473:pathology* / Q000532:radiotherapy; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D002467:Cell Nucleus / Q000235:genetics / Q000473:pathology; D015179:Colorectal Neoplasms / Q000473:pathology* / Q000532:radiotherapy; D005260:Female; D006023:Glycoproteins / Q000378:metabolism; D006570:Heterochromatin / Q000378:metabolism*; D006657:Histones / Q000378:metabolism; D006801:Humans; D008297:Male; D008745:Methylation; D051379:Mice; D008819:Mice, Nude; D008875:Middle Aged; D009368:Neoplasm Transplantation; D014411:Neoplastic Stem Cells / Q000473:pathology / Q000528:radiation effects*; D010315:Particle Accelerators; D010455:Peptides / Q000378:metabolism; D011829:Radiation Dosage; D011836:Radiation Tolerance
D000071916:AC133 Antigen; D015703:Antigens, CD; D006023:Glycoproteins; D006570:Heterochromatin; D006657:Histones; C000606023:PROM1 protein, human; D010455:Peptides; C000606024:Prom1 protein, mouse