[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.

CONCLUSION:

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: Fang Liu, Lingyan Wang, Jieyu Li, Shuping Chen, Yunbin Ye
• Journal: Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology (Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi) Vol. 30 Issue 7 Pg. 691-5 (Jul 2014) ISSN: 1007-8738 [Print] China
• PMID: 25001930 (Publication Type: English Abstract, Journal Article)

Chemical References

• CARD Signaling Adaptor Proteins
• EGR1 protein, human
• Early Growth Response Protein 1
• Transcription Factor RelA
• CARD11 protein, human
• Guanylate Cyclase

Topics

• Blotting, Western
• CARD Signaling Adaptor Proteins (genetics, metabolism)
• Cell Movement (genetics)
• Cell Proliferation
• Early Growth Response Protein 1 (genetics, metabolism)
• Guanylate Cyclase (genetics, metabolism)
• HEK293 Cells
• Humans
• K562 Cells
• Neoplasm Invasiveness
• Neoplasm Metastasis
• RNA Interference
• Reverse Transcriptase Polymerase Chain Reaction
• Signal Transduction (genetics)
• Transcription Factor RelA (genetics, metabolism)