PMID- 27911920
OWN - NLM
STAT- MEDLINE
DCOM- 20170630
LR  - 20231111
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 11
IP  - 12
DP  - 2016
TI  - SiRNA Targeting mTOR Effectively Prevents the Proliferation and Migration of 
      Human Lens Epithelial Cells.
PG  - e0167349
LID - 10.1371/journal.pone.0167349 [doi]
LID - e0167349
AB  - Posterior capsule opacification (PCO) is the most common complication that causes 
      visual decrease after extracapsular cataract surgery. The primary cause of PCO 
      formation is the proliferation of the residual lens epithelial cells (LECs). The 
      mammalian target of rapamycin (mTOR) plays an important role in the growth and 
      migration of LECs. In the current study, we used small interfering RNA (siRNA) to 
      specifically attenuate mTOR in human lens epithelial B3 cells (HLE B3). We aimed 
      to examine the effect of mTOR-siRNA on the proliferation, migration and 
      epithelial-to-mesenchymal transition (EMT) of HLE B3 cells and explore the 
      underlying mechanisms. The mTOR-siRNA was transfected into HLE B3 cells using 
      lipofectamine 2000. The mRNA and protein levels of mTOR were examined to confirm 
      the efficiency of mTOR-siRNA. The levels of mRNA and protein as well as the 
      activity of mTOR down-stream effectors p70 ribosomal protein S6 kinase (p70S6K) 
      and protein kinase B (PKB, AKT) were examined using real-time PCR or Western 
      blot, respectively. The cell proliferation was determined using cell counting kit 
      (CCK) 8 and cell growth curve assay. The cell migration was examined using 
      Transwell system and Scratch assay. MTOR-siRNA effectively eliminated mTOR mRNA 
      and protein. The proliferation and migration were significantly suppressed by 
      mTOR-siRNA transfection. mTOR-siRNA reduced the mRNA of p70S6K and AKT in a 
      time-dependent manner. Furthermore, the phosphorylation of p70S6K and AKT was 
      decreased by mTOR-siRNA. MTOR-siRNA also eliminated the formation of mTORC1 and 
      mTORC2 protein complex and blocked the transforming growth factor (TGF)-beta-induced 
      EMT. Our results suggested that mTOR-siRNA could effectively inhibit the 
      proliferation, migration and EMT of HLE B3 cells through the inhibition of p70S6K 
      and AKT. These results indicated that mTOR-siRNA might be an effective agent 
      inhibiting HLE cells growth and EMT following cataract surgery and provide an 
      alternative therapy for preventing PCO.
FAU - Zhang, Chunmei
AU  - Zhang C
AD  - Department of Ophthalmology, the First Affiliated Hospital of Harbin Medical 
      University, Harbin, P.R. China.
FAU - Liu, Jingjing
AU  - Liu J
AD  - Department of Ophthalmology, the First Affiliated Hospital of Harbin Medical 
      University, Harbin, P.R. China.
FAU - Jin, Na
AU  - Jin N
AD  - Department of Ophthalmology, the First Affiliated Hospital of Harbin Medical 
      University, Harbin, P.R. China.
FAU - Zhang, Guiming
AU  - Zhang G
AD  - Department of Ophthalmology, No.2 Hospital of Xiamen, Fujian, P.R. China.
FAU - Xi, Yahui
AU  - Xi Y
AD  - Department of Ophthalmology, the First Affiliated Hospital of Harbin Medical 
      University, Harbin, P.R. China.
FAU - Liu, Hongling
AU  - Liu H
AD  - Department of Ophthalmology, the First Affiliated Hospital of Harbin Medical 
      University, Harbin, P.R. China.
LA  - eng
PT  - Journal Article
DEP - 20161202
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (Multiprotein Complexes)
RN  - 0 (RNA, Small Interfering)
RN  - EC 2.7.1.1 (MTOR protein, human)
RN  - EC 2.7.11.1 (Mechanistic Target of Rapamycin Complex 1)
RN  - EC 2.7.11.1 (Proto-Oncogene Proteins c-akt)
RN  - EC 2.7.11.1 (Ribosomal Protein S6 Kinases, 70-kDa)
RN  - EC 2.7.11.1 (TOR Serine-Threonine Kinases)
SB  - IM
MH  - Capsule Opacification/genetics/metabolism/pathology
MH  - Cell Line
MH  - Cell Movement/*drug effects
MH  - Cell Proliferation/*drug effects
MH  - Epithelial Cells/*metabolism/pathology
MH  - Epithelial-Mesenchymal Transition/drug effects/genetics
MH  - Gene Expression Regulation/drug effects/genetics
MH  - Humans
MH  - Lens Capsule, Crystalline/*metabolism/pathology
MH  - Mechanistic Target of Rapamycin Complex 1
MH  - Multiprotein Complexes/genetics/metabolism
MH  - Proto-Oncogene Proteins c-akt/genetics/metabolism
MH  - RNA, Small Interfering/genetics/*pharmacology
MH  - Ribosomal Protein S6 Kinases, 70-kDa/genetics/metabolism
MH  - TOR Serine-Threonine Kinases/*biosynthesis/genetics/metabolism
PMC - PMC5135089
COIS- The authors have declared that no competing interests exist.
EDAT- 2016/12/03 06:00
MHDA- 2017/07/01 06:00
PMCR- 2016/12/02
CRDT- 2016/12/03 06:00
PHST- 2016/06/29 00:00 [received]
PHST- 2016/11/12 00:00 [accepted]
PHST- 2016/12/03 06:00 [entrez]
PHST- 2016/12/03 06:00 [pubmed]
PHST- 2017/07/01 06:00 [medline]
PHST- 2016/12/02 00:00 [pmc-release]
AID - PONE-D-16-26056 [pii]
AID - 10.1371/journal.pone.0167349 [doi]
PST - epublish
SO  - PLoS One. 2016 Dec 2;11(12):e0167349. doi: 10.1371/journal.pone.0167349. 
      eCollection 2016.