TY - JOUR
T1 - Upregulation of BMI1-suppressor miRNAs (miR-200c, miR-203) during terminal differentiation of colon epithelial cells
AU - Hisamori, Shigeo
AU - Mukohyama, Junko
AU - Koul, Sanjay
AU - Hayashi, Takanori
AU - Rothenberg, Michael Evan
AU - Maeda, Masao
AU - Isobe, Taichi
AU - Valencia Salazar, Luis Enrique
AU - Qian, Xin
AU - Johnston, Darius Michael
AU - Qian, Dalong
AU - Lao, Kaiqin
AU - Asai, Naoya
AU - Kakeji, Yoshihiro
AU - Gennarino, Vincenzo Alessandro
AU - Sahoo, Debashis
AU - Dalerba, Piero
AU - Shimono, Yohei
N1 - Publisher Copyright:
© 2022, Japanese Society of Gastroenterology.
PY - 2022/6
Y1 - 2022/6
N2 - Background: MicroRNAs (miRNAs) are key regulators of stem cell functions, including self-renewal and differentiation. In this study, we aimed to identify miRNAs that are upregulated during terminal differentiation in the human colon epithelium, and elucidate their role in the mechanistic control of stem cell properties. Methods: “Bottom-of-the-crypt” (EPCAM+/CD44+/CD66alow) and “top-of-the-crypt” (EPCAM+/CD44neg/CD66ahigh) epithelial cells from 8 primary colon specimens (6 human, 2 murine) were purified by flow cytometry and analyzed for differential expression of 335 miRNAs. The miRNAs displaying the highest upregulation in “top-of-the-crypt” (terminally differentiated) epithelial cells were tested for positive correlation and association with survival outcomes in a colon cancer RNA-seq database (n = 439 patients). The two miRNAs with the strongest “top-of-the-crypt” expression profile were evaluated for capacity to downregulate self-renewal effectors and inhibit in vitro proliferation of colon cancer cells, in vitro organoid formation by normal colon epithelial cells and in vivo tumorigenicity by patient-derivedxenografts (PDX). Results: Six miRNAs (miR-200a, miR-200b, miR-200c, miR-203, miR-210, miR-345) were upregulated in “top-of-the-crypt” cells and positively correlated in expression among colon carcinomas. Overexpression of the three miRNAs with the highest inter-correlation coefficients (miR-200a, miR-200b, miR-200c) associated with improved survival. The top two over-expressed miRNAs (miR-200c, miR-203) cooperated synergistically in suppressing expression of BMI1, a key regulator of self-renewal in stem cell populations, and in inhibiting proliferation, organoid-formation and tumorigenicity of colon epithelial cells. Conclusion: In the colon epithelium, terminal differentiation associates with the coordinated upregulation of miR-200c and miR-203, which cooperate to suppress BMI1 and disable the expansion capacity of epithelial cells.
AB - Background: MicroRNAs (miRNAs) are key regulators of stem cell functions, including self-renewal and differentiation. In this study, we aimed to identify miRNAs that are upregulated during terminal differentiation in the human colon epithelium, and elucidate their role in the mechanistic control of stem cell properties. Methods: “Bottom-of-the-crypt” (EPCAM+/CD44+/CD66alow) and “top-of-the-crypt” (EPCAM+/CD44neg/CD66ahigh) epithelial cells from 8 primary colon specimens (6 human, 2 murine) were purified by flow cytometry and analyzed for differential expression of 335 miRNAs. The miRNAs displaying the highest upregulation in “top-of-the-crypt” (terminally differentiated) epithelial cells were tested for positive correlation and association with survival outcomes in a colon cancer RNA-seq database (n = 439 patients). The two miRNAs with the strongest “top-of-the-crypt” expression profile were evaluated for capacity to downregulate self-renewal effectors and inhibit in vitro proliferation of colon cancer cells, in vitro organoid formation by normal colon epithelial cells and in vivo tumorigenicity by patient-derivedxenografts (PDX). Results: Six miRNAs (miR-200a, miR-200b, miR-200c, miR-203, miR-210, miR-345) were upregulated in “top-of-the-crypt” cells and positively correlated in expression among colon carcinomas. Overexpression of the three miRNAs with the highest inter-correlation coefficients (miR-200a, miR-200b, miR-200c) associated with improved survival. The top two over-expressed miRNAs (miR-200c, miR-203) cooperated synergistically in suppressing expression of BMI1, a key regulator of self-renewal in stem cell populations, and in inhibiting proliferation, organoid-formation and tumorigenicity of colon epithelial cells. Conclusion: In the colon epithelium, terminal differentiation associates with the coordinated upregulation of miR-200c and miR-203, which cooperate to suppress BMI1 and disable the expansion capacity of epithelial cells.
KW - BMI1
KW - Cell differentiation
KW - Colon
KW - Tumor-suppressor
KW - miRNA
UR - http://www.scopus.com/inward/record.url?scp=85125582038&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85125582038&partnerID=8YFLogxK
U2 - 10.1007/s00535-022-01865-9
DO - 10.1007/s00535-022-01865-9
M3 - Article
C2 - 35244768
AN - SCOPUS:85125582038
SN - 0944-1174
VL - 57
SP - 407
EP - 422
JO - Journal of Gastroenterology
JF - Journal of Gastroenterology
IS - 6
ER -
