A Cancer Reprogramming Method Using MicroRNAs as a Novel Therapeutic Approach against Colon Cancer: Research for Reprogramming of Cancer Cells by MicroRNAs.
BACKGROUND
We previously generated induced pluripotent stem cells by reprograming adipose stem cells through the introduction of microRNAs targeting four transcription factors (Oct3/4, Sox2, c-Myc, and Klf4). In this study, we aimed to reprogram cancer cells using microRNAs to explore their therapeutic potential.


METHODS
Mature microRNAs (mir-302a-d, 369-3p and 5p, and mir-200c, as needed) were introduced into colon cancer cells (DLD-1, RKO, and HCT116) using lipofection.


RESULTS
The transfected cells exhibited an embryonic stem cell-like morphology and expressed the undifferentiated marker genes Nanog, Oct3/4, SOX2, and Klf4, as well as tumor-related antigen-1-60. These cells expressed neurogenic or adipogenic markers, indicating that reprogramming of the cancer cells was partially successful. Moreover, we found that miRNA-expressing DLD-1 cells showed low proliferative activity in vitro and in vivo, accompanied by increased expression of the tumor suppressor genes p16 (ink4a) and p21 (waf1) . miRNA-expressing DLD-1 cells also exhibited enhanced sensitivity to 5-fluorouracil, possibly through the downregulation of multidrug-resistant protein 8. The reprogrammed cells from DLD-1, RKO, and HCT116 cells exhibited reduced c-Myc expression, in contrast to the high c-Myc expression in the induced pluripotent cancer cells that were generated using four transcription factors.


CONCLUSIONS
Our cancer reprogramming method employing simple lipofection of mature microRNAs is safe and well suited for clinical application, because it avoids integration of exogenous genes into the host genome and allows escape from augmentation of c-Myc gene expression.
Department of Gastroenterological Surgery, Graduate School of Medicine, Osaka University, Osaka, Japan.
Susumu
S
Miyazaki
Department of Gastroenterological Surgery, Graduate School of Medicine, Osaka University, Osaka, Japan. hyamamoto@gesurg.med.osaka-u.ac.jp.
Hirofumi
H
Yamamoto
Department of Surgery, Osaka Medical Center for Cancer and Cardiovascular Diseases, Osaka, Japan.
Norikatsu
N
Miyoshi
Department of Gastroenterological Surgery, Graduate School of Medicine, Osaka University, Osaka, Japan.
Xin
X
Wu
Department of Gastroenterological Surgery, Graduate School of Medicine, Osaka University, Osaka, Japan.
Hisataka
H
Ogawa
Department of Gastroenterological Surgery, Graduate School of Medicine, Osaka University, Osaka, Japan.
Mamoru
M
Uemura
Department of Gastroenterological Surgery, Graduate School of Medicine, Osaka University, Osaka, Japan.
Junichi
J
Nishimura
Department of Gastroenterological Surgery, Graduate School of Medicine, Osaka University, Osaka, Japan.
Taishi
T
Hata
Department of Gastroenterological Surgery, Graduate School of Medicine, Osaka University, Osaka, Japan.
Ichiro
I
Takemasa
Department of Gastroenterological Surgery, Graduate School of Medicine, Osaka University, Osaka, Japan.
Tsunekazu
T
Mizushima
Department of Frontier Science for Cancer and Chemotherapy, Graduate School of Medicine, Osaka University, Osaka, Japan.
Masamitsu
M
Konno
Department of Gastroenterological Surgery, Graduate School of Medicine, Osaka University, Osaka, Japan.
Yuichiro
Y
Doki
Department of Gastroenterological Surgery, Graduate School of Medicine, Osaka University, Osaka, Japan.
Masaki
M
Mori
Department of Frontier Science for Cancer and Chemotherapy, Graduate School of Medicine, Osaka University, Osaka, Japan.|Department of Cancer Profiling Discovery, Graduate School of Medicine, Osaka University, Osaka, Japan.
Hideshi
H
Ishii
D000818:Animals; D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D002454:Cell Differentiation / Q000187:drug effects; D019070:Cell Lineage / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D003110:Colonic Neoplasms / Q000188:drug therapy / Q000235:genetics* / Q000473:pathology; D004353:Drug Evaluation, Preclinical; D053595:Embryonic Stem Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D005472:Fluorouracil / Q000494:pharmacology; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D000090062:Kruppel-Like Factor 4; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D035683:MicroRNAs / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured
D000964:Antimetabolites, Antineoplastic; C000715467:KLF4 protein, human; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D035683:MicroRNAs; D012333:RNA, Messenger; D005472:Fluorouracil