Tuftelin Is Required for NGF-Induced Differentiation of PC12 Cells

被引:13
作者
Shilo, Dekel [1 ,2 ]
Cohen, Gadi [3 ]
Blumenfeld, Anat [1 ]
Goren, Koby [1 ]
Hanhan, Salem [1 ]
Sharon, Shay [1 ]
Haze, Amir [1 ]
Deutsch, Dan [1 ]
Lazarovici, Philip [3 ]
机构
[1] Hebrew Univ Jerusalem, Inst Dent Sci, Fac Med Dent, Dent Res Lab, Jerusalem, Israel
[2] Rambam Hlth Care Campus, 8 HaAliyah St, IL-35254 Haifa, Israel
[3] Hebrew Univ Jerusalem, Inst Drug Res, Sch Pharm, Fac Med, Jerusalem, Israel
关键词
Nerve growth factor; NGF; PC12; cells; Cell differentiation; Neuronal outgrowth; Tuftelin; Tuft1; NERVE GROWTH-FACTOR; GENE-EXPRESSION; MESSENGER-RNAS; IDENTIFICATION; HYPOXIA; PROTEIN; TRKA;
D O I
10.1007/s12031-019-01292-1
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Nerve growth factor (NGF) promotes pleiotropic gene transcription-dependent biological effects, in neuronal and non-neuronal cells, including survival, proliferation, differentiation, neuroprotection, pain, and angiogenesis. It is hypothesized that during odontogenesis, NGF may be implicated in morphogenetic and mineralization events by affecting proliferation and/or differentiation of dental cells. Tuftelin belongs to the enamel associated teeth proteins and is thought to play a role in enamel mineralization. We previously reported that tuftelin transcript and protein, which are ubiquitously expressed in various tissues of embryos, adults, and tumors, were significantly upregulated during NGF-induced PC12 differentiation. To further confirm the involvement of tuftelin in the differentiation process, we established a tuftelin-knockdown neuronal PC12 cell model, using a non-cytotoxic siRNA directed towards sequences at the 3 UTR of the tuftelin gene. Using real-time PCR, we quantified tuftelin mRNA expression and found that tuftelin siRNA, but not scrambled siRNA or transfection reagents, efficiently depleted about 60% of NGF-induced tuftelin mRNA transcripts. The effect of tuftelin siRNA was quantified up to 6days of NGF-induced differentiation. Using immunofluorescence and western blot analyses, we also found a direct correlation between reduction of 60-80% in tuftelin protein expression and inhibition of about 50-70% in NGF-induced differentiation of the cells, as was detected after 3-6days of treatment. These results demonstrate an important role for tuftelin in NGF-induced differentiation of PC12 cells. Tuftelin could be a useful target for drug development in disease where neurotrophin therapy is required.
引用
收藏
页码:135 / 143
页数:9
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