TY - JOUR
T1 - MicroRNA-141-3p regulates cellular proliferation, migration, and invasion in esophageal cancer by targeting tuberous sclerosis complex 1
AU - Phatak, Pornima
AU - Noe, Michael
AU - Asrani, Kaushal
AU - Chesnick, Ingrid E.
AU - Greenwald, Bruce D.
AU - Donahue, James M.
N1 - Funding Information:
The authors thank Dr Jeffrey T. Mason for providing the droplet digital PCR system and Dr Paul W. Sanders for endowing the Plate reader. They also thank Ms E. Parker for her assistance in compiling the references and Mr. Damian Schwartz for his aassistance in lab administration. This work was supported by the Department of Veterans Affairs, U.S.A. Merit Award (to JMD; 5101CX001434‐02).
Publisher Copyright:
© 2020 Wiley Periodicals LLC
PY - 2021/2
Y1 - 2021/2
N2 - MicroRNA (miR)−141-3p, which functions as an oncogene in multiple malignancies, has been shown to be highly overexpressed in esophageal cancer cells in our previous work. miR-141-3p is predicted to bind the messenger RNA (mRNA) of tuberous sclerosis complex 1 (TSC1), a tumor suppressor, with high affinity. In this study, we investigated the expression and functional interaction between miR-141-3p and TSC1 in esophageal cancer cells. Experiments were conducted in four esophageal cancer lines and in tumor cells isolated from human esophageal cancer specimens by laser capture microdissection. miR-141-3p expression was measured by real time and droplet digital PCR. Biotinylated RNA pull-down and luciferase reporter assays were used to assess binding. miR-141-3p function was tested by assessing proliferation, migration, invasion, and induction of autophagy following its silencing. We found that miR-141-3p levels were increased in TE7, OE33, and TE10 esophageal cancer cells compared to FLO-1 cells, with similar heterogeneity observed in human esophageal cancer specimens. Silencing of miR-141-3p led to increased TSC1 protein expression in these cells and was associated with increased TSC1 translation. Binding studies reveal that miR-141-3p binds to each of the predicted binding sites in the 3′-untranslated region of TSC1 mRNA. Following miR-141-3p silencing, TE7, OE33, and TE10 cells exhibited decreased proliferation, migration, and invasion, as well as enhanced autophagy. Importantly, these phenotypic effects were replicated by overexpression of TSC1 alone in these cells. Our results indicate that miR-141-3p functions in an oncogenic capacity in a subset of esophageal cancer cells, in part by suppressing TSC1 expression.
AB - MicroRNA (miR)−141-3p, which functions as an oncogene in multiple malignancies, has been shown to be highly overexpressed in esophageal cancer cells in our previous work. miR-141-3p is predicted to bind the messenger RNA (mRNA) of tuberous sclerosis complex 1 (TSC1), a tumor suppressor, with high affinity. In this study, we investigated the expression and functional interaction between miR-141-3p and TSC1 in esophageal cancer cells. Experiments were conducted in four esophageal cancer lines and in tumor cells isolated from human esophageal cancer specimens by laser capture microdissection. miR-141-3p expression was measured by real time and droplet digital PCR. Biotinylated RNA pull-down and luciferase reporter assays were used to assess binding. miR-141-3p function was tested by assessing proliferation, migration, invasion, and induction of autophagy following its silencing. We found that miR-141-3p levels were increased in TE7, OE33, and TE10 esophageal cancer cells compared to FLO-1 cells, with similar heterogeneity observed in human esophageal cancer specimens. Silencing of miR-141-3p led to increased TSC1 protein expression in these cells and was associated with increased TSC1 translation. Binding studies reveal that miR-141-3p binds to each of the predicted binding sites in the 3′-untranslated region of TSC1 mRNA. Following miR-141-3p silencing, TE7, OE33, and TE10 cells exhibited decreased proliferation, migration, and invasion, as well as enhanced autophagy. Importantly, these phenotypic effects were replicated by overexpression of TSC1 alone in these cells. Our results indicate that miR-141-3p functions in an oncogenic capacity in a subset of esophageal cancer cells, in part by suppressing TSC1 expression.
KW - esophageal cancer
KW - miR-141-3p
KW - tuberous sclerosis complex 1
UR - http://www.scopus.com/inward/record.url?scp=85098446863&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85098446863&partnerID=8YFLogxK
U2 - 10.1002/mc.23274
DO - 10.1002/mc.23274
M3 - Article
C2 - 33382472
AN - SCOPUS:85098446863
SN - 0899-1987
VL - 60
SP - 125
EP - 137
JO - Molecular Carcinogenesis
JF - Molecular Carcinogenesis
IS - 2
ER -