Comparison On Effects Of EGF, TNF-α,IL-1α, And IL-10 On The Invasiveness Between Human Trophoblastic Cells And Ovarian Cancer Cell Line HO-8910

Objective:To investigate effects of EGF, TNF- a , IL-1 a , and IL-10 on the invasiveness of primary cultured human trophoblastic cells and ovarian cancer cell line HO-8910 in vitro. To compare the mRNA experssion, the protein content of the genes associated with invasion, and the number of the cells which invaded through the Matrigel Matrix covered on the membrane with 8 micron pores. Methods:Samples of first-trimester placentae, provided by department of obstetrics and gynecology in Nanfang Hospital, were obtained from women who were at elective termination of selected pregnancies performed by evacuation between the 6th and 8th weeks, with the permission of the local ethical committee. Tissue-culture method was used to get the primary human trophoblastic cells of first trimester pregnancy. The trophoblastic cells and their purity were identified by immunocytochemistry with specific monoclonal antibodies. The cell line HO-8910 was cultured in vitro. Then the cells were incubated with EGF, TNF- a , IL-1 a , and IL-10 for 48 hours in vitro, respectively. Levels of the mRNA expression of MMP-2, MMP-9, MTA1, and KAI1 was detected by RT-PCR method. Cellular ELISA was applied to detecte the protein content of MMP-2 and CD44v6. The invading cells through the matrigel were stained and counted under microscope. All data were analyzed by statistical analysis. Results:1. MTA1 mRNA and KAI1 mRNA were detected in human trophoblasticcell by RT-PCR, and CD44v6 was also found to express in human trophoblastic cell by cellular ELISA.2. EGF significantly increased the expression of MMP-2 mRNA and MMP-9 mRNA in these two kinds of the cells at dose-dependent; TNF- a also significantly increased the expression of MMP-2 mRNA in human trophoblastic cell and the cell line HO-8910, but only up-regulated MMP-9 mRNA in the cell line. And the effect decreased as the concentration of TNF-a increased. IL-1 a and IL-10 reduced MMP-2 mRNA content in human trophoblast, but increased the expression of MMP-2 mRNA and MMP-9 mRNA in the cell line. All cytokines used inhibited MAI1 mRNA and KAI1 mRNA expression, except that EGF promoted KAI1 mRNA expression in human trophoblastic cell.3. Both EGF and TNF-a increased MMP-2 and CD44v6 expression in human trophoblast, but IL-1 a and IL-10 reduced them. In the cell line HO-8910, MMP-2 was up-regulated by EGF and down-regulated by IL-10, EGF, TNF- a , and IL-1 a promoted CD44v6 expression.4. The number of invading human trophoblastic cells was greater than that of the cell line HO-8910 cultured in normal medium. TNF- a could restrain the invading cell number of either human trophoblast or the cell line when its concentration increased. EGF could significantly increase the invading number of the cell line. And a dose-dependent effect existed. Conclusions:1. Human trophoblastic cell could express MTA1 mRNA, KAI1 mRNA, and CD44v6. It suggested that MTA1, KAI1, and CD44v6 participate in the regulation of human trophoblastic cell invasion. All of these showed the similarity of invasiveness between human trophoblastic cells and invasive tumor cells.2. EGF, TNF- a , IL-1 a , and IL-10 would have different effects on the regulation of invasion for human trophoblastic cell and the cell line HO-8910.3. The invasiveness of human trophoblastic cell would be stronger thanthat of the ovarian cancer cell line HO-8910.