title: [Knock-down of CARMA1 in K562 cells inhibits the invasion and metastasis of K562 cells].
abstract: OBJECTIVE
  To study the effects of caspase recruitment domain membrane-associated guanylate kinase protein 1 (CARMA1) knock-down using RNAi technology on cell proliferation, colony forming, invasion and metastasis of the K562 cells.
  
  
  METHODS
  K562 cells with stably-silenced CARMA1 gene was constructed by lentivirus-mediated RNAi technology. CARMA1 mRNA and protein levels were respectively detected by reverse transcription PCR (RT-PCR) and Western blotting. Cell proliferation was analyzed by trypan blue exclusion; colony forming was detected by colony forming assay; invasion and metastasis in vitro were determined with Transwell® assay with or without matrigel. Signaling pathway molecules were detected by RT-PCR and Western blotting.
  
  
  RESULTS
  Six cell lines with knocked-down gene were successfully constructed. Of them, one was negative control (K562/sh-eGFP), and the other five were CARMA1-silenced cells. K562/shCARMA 1-93 showed the greatest inhibition of CARMA 1 gene and protein expressions. Trypan blue exclusion and colony forming assay showed that the ability of cell proliferation and colony forming were significantly suppressed in K562/shCARMA 1-93 (P<0.01). Compared with blank control K562 and negative control K562/sh-eGFP, the growth inhibition rates were respectively 29.3% and 28.6%; the colony forming inhibition rates were respectively 37.6% and 34.1%; Transwell® assay with or without matrigel showed that the number of K562/shCARMA1-93 which passed through the membrane was obviously lower than that of the control group (P<0.01). The expressions of nucleic transcription factor-κB (NF-κB) and early growth response 1 (EGR1) were reduced following the inhibition of CARMA1.
  
  
  CONCLUSIONS
  CARMA1 gene knock-down had an impact on cell proliferation, colony forming, invasion and metastasis of K562 cells, which is possibly related to the inhibition of NF-κB and JNK/EGR-1 signaling pathways.
authors.affiliation: Immuno-Oncology Laboratory, Fujian Provincial Cancer Hospital, Teaching Hospital, Fujian Medical University; Fujian Provincial Key Laboratory of Translational Cancer Medicine, Fuzhou 350014, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Liu
authors.affiliation: Immuno-Oncology Laboratory, Fujian Provincial Cancer Hospital, Teaching Hospital, Fujian Medical University; Fujian Provincial Key Laboratory of Translational Cancer Medicine, Fuzhou 350014, China.
authors.forename: Lingyan
authors.initials: L
authors.lastname: Wang
authors.affiliation: Immuno-Oncology Laboratory, Fujian Provincial Cancer Hospital, Teaching Hospital, Fujian Medical University; Fujian Provincial Key Laboratory of Translational Cancer Medicine, Fuzhou 350014, China.
authors.forename: Jieyu
authors.initials: J
authors.lastname: Li
authors.affiliation: Immuno-Oncology Laboratory, Fujian Provincial Cancer Hospital, Teaching Hospital, Fujian Medical University; Fujian Provincial Key Laboratory of Translational Cancer Medicine, Fuzhou 350014, China.
authors.forename: Shuping
authors.initials: S
authors.lastname: Chen
authors.affiliation: Immuno-Oncology Laboratory, Fujian Provincial Cancer Hospital, Teaching Hospital, Fujian Medical University; Fujian Provincial Key Laboratory of Translational Cancer Medicine, Fuzhou 350014, China.
authors.forename: Yunbin
authors.initials: Y
authors.lastname: Ye
mesh_terms: D015153:Blotting, Western; D053468:CARD Signaling Adaptor Proteins / Q000235:genetics* / Q000378:metabolism; D002465:Cell Movement / Q000235:genetics*; D049109:Cell Proliferation; D051766:Early Growth Response Protein 1 / Q000235:genetics / Q000378:metabolism; D006162:Guanylate Cyclase / Q000235:genetics* / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D020014:K562 Cells; D009361:Neoplasm Invasiveness; D009362:Neoplasm Metastasis; D034622:RNA Interference; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000235:genetics; D051996:Transcription Factor RelA / Q000235:genetics / Q000378:metabolism
chemical_list: D053468:CARD Signaling Adaptor Proteins; C497325:EGR1 protein, human; D051766:Early Growth Response Protein 1; D051996:Transcription Factor RelA; C426242:CARD11 protein, human; D006162:Guanylate Cyclase