Synchronization of Plodia interpunctella lepidopteran cells and effects of 20-hydroxyecdysone

被引：7

作者：

Siaussat, D ^{[1
]}

Mottier, V ^{[1
]}

Bozzolan, F ^{[1
]}

Porcheron, P ^{[1
]}

Debernard, S ^{[1
]}

机构：

[1] Univ Paris 06, Lab Physiol Cellulaire Invertebras, F-75005 Paris, France

来源：

INSECT MOLECULAR BIOLOGY | 2004年 / 13卷 / 02期

关键词：

Plodia cell cycle; hydroxyurea; synchronization; Plodia HR3 and E75; 20-hydroxyecdysone;

D O I：

10.1111/j.0962-1075.2004.00475.x

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：

We have investigated the molecular and cellular mechanisms involved in the control of insect cell cycle by 20-hydroxyecdysone (20E) using the IAL-PID2 cell line established from imaginal wing discs of Plodia interpunctella. We first defined conditions for use of hydroxyurea, a reversible inhibitor of DNA synthesis, in order to synchronize the IAL-PID2 cells in their division cycle. A high degree of synchrony was reached when cells were exposed to two consecutive hydroxyurea treatments at 1 mM for 36 h spaced 16 h apart. Under these conditions, flow cytometry analysis demonstrated that 20E at 10(-6) M induced an inhibition of cell growth by an arrest of 90% of the cells in G2/M phase. Using cDNA probes specifically designed from E75 and HR3 nuclear receptors of Plodia interpunctella, we showed that PiE75 and PHR3 were highly induced by 20E through S and G2 phases with maximal enhancement just before the G2/M arrest of cells. These findings suggest that PiE75 and PHR3 could be involved in a 20E-induced genetic cascade leading to G2/M arrest.

引用

页码：179 / 187

页数：9

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