Induction of G1 cell cycle arrest and P15INK4b expression by ECRG1 through interaction with Miz-1.

ECRG1 is a novel candidate of tumor suppressor gene identified from human esophagus. To study the biological role of ECRG1 gene, we performed a GAL4-based yeast two-hybrid screen of a human fetal liver cDNA library. Using the ECRG1 cDNA as bait, we identified two putative clones as associated proteins, Miz-1 and FLNA (Filamin A). The interaction of ECRG1 and Miz-1 was confirmed by glutathione-S-transferase (GST)-pull-down assays in vitro and co-immunoprecipitation experiments in vivo. ECRG1 was co-localized with Miz-1 in nucleus, as shown by confocal microscopy. Transfection of ECRG1 gene into the esophageal cancer (EC) cells inhibited cell proliferation and induced G1 phase arrest of cell cycle. In the co-transfection of ECRG1 and Miz-1 assays, we found inhibition of cell proliferation and G1/S phase in EC cells, but the levels of cell proliferation inhibition and G1/S phase arrest were more strongly compared with the transfection of ECRG1 or Miz-1 alone. In addition, the interaction of ECRG1 and Miz-1 could induce expression of P15(INK4b) gene in esophageal cancer 9706 (EC9706) cells. However, the transfection of ECRG1 or Miz-1 alone was not revealed the expressions of P15(INK4b) gene. When antisense ECRG1 interdicted expression of endogenous ECRG1 in Balb/c-3T3 cells, Transfection of Miz-1 couldn't induce P15(INK4b) expression. The results provide evidences that ECRG1 and Miz-1 in EC cells may be acting as a co-functional protein associated with regulation of cell cycle and induction of P15(INK4b) expression. It suggests that ECRG1 may inhibit tumor cell growth by affecting cell cycle, and that expression of P15(INK4b) may be likely to enhance G1 cell cycle arrest during the interaction of ECRG1 and Miz-1. The physical interaction of ECRG1 and Miz-1 may play an important role in carcinogenesis of EC.

Zhao N ，Wang J ，Cui Y ，Guo L ，Lu SH ... -  《JOURNAL OF CELLULAR BIOCHEMISTRY》

[Search for interacting proteins of esophageal cancer related gene-1 encoded protein through the yeast two-hybrid system].

Wang J ，Fan Y ，Guo L ，Lu S ... -  《-》

Repression of p15INK4b expression by Myc through association with Miz-1.

Staller P ，Peukert K ，Kiermaier A ，Seoane J ，Lukas J ，Karsunky H ，Möröy T ，Bartek J ，Massagué J ，Hänel F ，Eilers M ... -  《NATURE CELL BIOLOGY》

Suppression of tumorigenesis and induction of p15(ink4b) by Smad4/DPC4 in human pancreatic cancer cells.

Peng B ，Fleming JB ，Breslin T ，Grau AM ，Fojioka S ，Abbruzzese JL ，Evans DB ，Ayers D ，Wathen K ，Wu T ，Robertson KD ，Chiao PJ ... -  《CLINICAL CANCER RESEARCH》

Consistent inactivation of p19(Arf) but not p15(Ink4b) in murine myeloid cells transformed in vivo by deregulated c-Myc.

Haviernik P ，Schmidt M ，Hu X ，Wolff L ... -  《ONCOGENE》