Pankaj Suman Ph.D., Nachiket Shembekar M.Sc., Satish Kumar Gupta Ph.D.
Volume 99, Issue 2, Pages 533-542.e2, February 2013
To identify the invasion associated molecules during leukemia inhibitory factor (LIF) mediated increase in the invasion of trophoblast cells.
Cultured trophoblastic HTR-8/SVneo cells were treated with LIF.
Main Outcome Measure(s):
Matrigel matrix based invasion assay of HTR-8/SVneo cells. Signaling molecules associated with LIF mediated increase in invasion were investigated by Western blot, cDNA microarray, qRT-PCR, immunofluorescence, immunohistochemistry and gene silencing by siRNA.
Treatment of HTR-8/SVneo cells with LIF (50 ng/ml) led to a significant increase in invasion. Treatment with LIF also led to an increase in nuclear localization of activated STAT1 and STAT3. Amongst 237 differentially expressed genes following LIF treatment, expression of pappalysin 1, SERPINB3, podoplanin, integrin β3, ID1, ICAM1 etc went up while, tissue inhibitor of matrix metalloproteinase 1 (TIMP1), TIMP2 and TIMP3 went down significantly. Presence of several of these proteins has also been demonstrated in human trophoblast cells. Silencing of pappalysin 1 led to a significant reduction in basal as well as LIF mediated invasiveness of HTR-8/SVneo cells.
Identification of novel molecules associated with LIF mediated increase in trophoblastic cell invasion may facilitate our understanding of implantation biology.
Read the full text at: http://www.fertstert.org/article/S0015-0282(12)02304-7/fulltext