PMID- 23643161
OWN - NLM
STAT- MEDLINE
DCOM- 20130730
LR  - 20211203
IS  - 1007-8738 (Print)
IS  - 1007-8738 (Linking)
VI  - 29
IP  - 4
DP  - 2013 Apr
TI  - [Effect of Kruppel like factor 4 gene silencing on apoptosis and phagocytosis of 
      murine RAW264.7 macrophages].
PG  - 341-4
AB  - OBJECTIVE: To investigate the effect of down-regulation of Kruppel like factor 4 
      (KLF4) by RNA interference on proliferation, apoptosis and phagocytosis in murine 
      RAW264.7 macrophage cell line. METHODS: Stable KLF4 silencing in RAW264.7 cells 
      was achieved by recombinant shRNA plasmid targeting murine KLF4 gene via liposome 
      mediated transfection, followed by G418 selection. The efficacy of KLF4 silencing 
      in G418 resistant cells was verified by fluorescent microcopy and Western 
      blotting, respectively. Proliferative activity was analyzed by CCK-8 assay. 
      Apoptosis and phagocytosis were evaluated by annexinV-FITC/PI staining and flow 
      cytometry with assistant use of fluorescein-labeled E.coli K-12 particles, 
      respectively. RESULTS: KLF4 protein expression was significantly down-regulated 
      by the recombinant shRNA plasmid as compared with negative control (NC) plasmid 
      transfection, inhibition rate being over 76%. From the third day, KLF4-silencing 
      cells exhibited lower proliferative activity as compared with NC RAW264.7 cells 
      (P<0.05). In resting state, apoptosis rate in wide type, NC and KLF4-silencing 
      cells were 1.73%, 6.85% and 12.76%, respectively, and KLF4-silencing resulted in 
      a statistical difference in apoptosis as compared with NC cells (P<0.05). 
      Finally, phagocytic capability in wide type, NC and KLF4-silencing cells revealed 
      by the mean fluorescence intensity of engulfed FITC-labeled E.coli particles were 
      122.0 +/- 2.80, 48.97 +/- 5.69 and 80.10 +/- 4.61, respectively, and compared with NC 
      cells, KLF4-silencing cells had a significant increase in phagocytosis (P<0.05). 
      CONCLUSION: KLF4 gene silencing inhibits RAW264.7 macrophage proliferation, 
      increase apoptosis and enhance phagocytic function in resting state, which 
      provides a novel tool for revealing the role of KLF4 in macrophage immunity.
FAU - Ji, Wenjie
AU  - Ji W
AD  - Department of Respiration and Critical Care Medicine, Pingjin Hospital, Logistics 
      University of Chinese People's Armed Police Forces, Tianjin 300162, China. 
      wenjieji@gmail.com
FAU - Chen, Xuefen
AU  - Chen X
FAU - Ma, Yongqiang
AU  - Ma Y
FAU - Hu, Daochuan
AU  - Hu D
FAU - Lu, Ruiyi
AU  - Lu R
FAU - Zhou, Xin
AU  - Zhou X
FAU - Wei, Luqing
AU  - Wei L
LA  - chi
PT  - Journal Article
PL  - China
TA  - Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi
JT  - Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular 
      immunology
JID - 101139110
RN  - 0 (Klf4 protein, mouse)
RN  - 0 (Kruppel-Like Factor 4)
RN  - 0 (Kruppel-Like Transcription Factors)
SB  - IM
MH  - Animals
MH  - Apoptosis/*genetics
MH  - Cell Growth Processes/physiology
MH  - Cell Line
MH  - Down-Regulation
MH  - Gene Silencing
MH  - Kruppel-Like Factor 4
MH  - Kruppel-Like Transcription Factors/*genetics
MH  - Macrophages/cytology/*physiology
MH  - Mice
MH  - Phagocytosis/*genetics
MH  - RNA Interference
EDAT- 2013/05/07 06:00
MHDA- 2013/07/31 06:00
CRDT- 2013/05/07 06:00
PHST- 2013/05/07 06:00 [entrez]
PHST- 2013/05/07 06:00 [pubmed]
PHST- 2013/07/31 06:00 [medline]
PST - ppublish
SO  - Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2013 Apr;29(4):341-4.