PMID- 17403315
OWN - NLM
STAT- MEDLINE
DCOM- 20081021
LR  - 20211203
IS  - 0376-2491 (Print)
IS  - 0376-2491 (Linking)
VI  - 87
IP  - 1
DP  - 2007 Jan 2
TI  - [Effect of RNA interference targeting mammalian target of rapamycin on the 
      proliferation of vascular smooth muscle cell and the intimal hyperplasia].
PG  - 58-62
AB  - OBJECTIVE: To construct RNA interference (RNAi) vector of mammalian target of 
      rapamycin (mTOR) and study the effect on the proliferation of vascular smooth 
      muscle cell (VSMC) and the intimal hyperplasia (IH). METHODS: The sequence of 
      short hairpin RNA (shRNA) targeting mTOR was designed and synthesized by chemical 
      method and inserted into a retroviral vector pLXIN, then the vector was packaged 
      in PT67 cells. The efficiency of inhibition was verified by Northern blot and 
      Western blot after transfection to VSMC. The changes of p70s 6k and 4E-BP1 were 
      also determined. The proliferation activity of VSMC was determined by 
      flowcytometry and MTT. Autogenous vein graft model were established in 54 rats. 
      Three groups were divided, including gene therapy group, control group and vein 
      graft group. The grafted veins were harvested 7 days, 14 days and 28 days after 
      transplanting respectively. The expression of mTOR was determined by 
      immunohistochemistry stain and Western blot. IH was also observed. The apoptotic 
      VSMC was determined by TUNEL. RESULTS: The shRNA fragments was inserted into 
      pLXIN vector. The shRNA vector was successfully constructed. Additionally, the 
      vector targeting mTOR gene could efficiently decrease the mRNA and protein 
      expression of mTOR and p70s 6k in VSMC, while the expression of 4E-BP1 increased 
      significantly. The cell cycle of VSMC was stunned in phase G(0)/G(1). The protein 
      expression of mTOR in vein graft was inhibited by transfection of pLXIN-shRNA 
      targeting mTOR (P < 0.01) and the IH was also inhibited (P < 0.01), but the 
      apoptosis of VSMC increased significantly (P < 0.01). CONCLUSION: The mTOR RNAi 
      retroviral vector has been constructed successfully, which can significantly 
      inhibit the proliferation of VSMC and the IH, and promote apoptosis after vein 
      grafting.
FAU - Hu, Xin-Hua
AU  - Hu XH
AD  - Department of Vascular Surgery, First Affiliated Hospital, China Medical 
      University, Shenyang, China. huxinhuacmu@hotmail.com
FAU - Zhang, Qiang
AU  - Zhang Q
FAU - Yang, Jun
AU  - Yang J
FAU - Liu, Cheng-Wei
AU  - Liu CW
FAU - Zhang, Zhi-Shen
AU  - Zhang ZS
FAU - Yang, De-Hua
AU  - Yang DH
LA  - chi
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - China
TA  - Zhonghua Yi Xue Za Zhi
JT  - Zhonghua yi xue za zhi
JID - 7511141
RN  - 0 (RNA, Small Interfering)
RN  - EC 2.7.- (Protein Kinases)
RN  - EC 2.7.11.1 (TOR Serine-Threonine Kinases)
SB  - IM
MH  - Animals
MH  - Apoptosis
MH  - Blood Vessels/transplantation
MH  - Blotting, Western
MH  - Cell Differentiation
MH  - *Cell Proliferation
MH  - Female
MH  - Genetic Therapy
MH  - Hyperplasia
MH  - Immunohistochemistry
MH  - Male
MH  - Muscle, Smooth, Vascular/cytology/*metabolism
MH  - Myocytes, Smooth Muscle/cytology/metabolism
MH  - Protein Kinases/genetics/*metabolism
MH  - *RNA Interference
MH  - RNA, Small Interfering/genetics
MH  - Rats
MH  - Rats, Wistar
MH  - TOR Serine-Threonine Kinases
MH  - Transfection
MH  - Tunica Intima/metabolism/*pathology/surgery
EDAT- 2007/04/04 09:00
MHDA- 2008/10/22 09:00
CRDT- 2007/04/04 09:00
PHST- 2007/04/04 09:00 [pubmed]
PHST- 2008/10/22 09:00 [medline]
PHST- 2007/04/04 09:00 [entrez]
PST - ppublish
SO  - Zhonghua Yi Xue Za Zhi. 2007 Jan 2;87(1):58-62.