Effects Of Lysophosphatidic Aicd On The Expression Of P120 CATENIN In Human Ovarian Cancer Cells

Objective:1. To investigate the effects of lysophosphatidic aicd (LPA) on the expression of P120 catenin(P120ctn) in human serous ovarian cancer SKOV3 cells.2. To study the reverse effect of Genistein as protein tyrosine kinase inhibitor on the expression of P120ctn in SKOV3 cells induced by LPA, and to approach the effectting mechanisms of LPA and P120ctn on the carcinogenesis, development, infiltrating and metastasis of ovarian cancer.Methods:1. After 6,12,24h treatting the SKOV3 cells in logarithmic growth phase with final concentration of 0, 0.1, 1, 10, 20, 40, 80μmol LPA /L and 0,10,20,50,100,200μmol Genistein /L, the proliferation rate and proliferation-inhibition rate of SKOV3 cells were detected by MTT assay.2. After 6,12,24h treatting the SKOV3 cells with 20μmol/L LPA and 200μmol/L Genistein, the expression of P120ctn in membrane and cytoplasm of SKOV3 cells was marked by double-label immunofluore- scence technique, and analysed by confocal laser scanning microscopy (CLSM); The changes of expression P120ctn was detected by RT-PCR and Western blot; The growth cycles of the SKOV3 cells was examined with Flow cytometry(FCM); Finally, the invasive and imgration ability of SKOV3 cells was detected by Millicell small chamber invasion assay in vitro.Results:1. The results of MTT showed that the proliferation of SKOV3 cells effected by 20μmol /L LPA for 24h reched a climax(P<0.05); but this effect of LPA could be obviously repressed by 200μmol/L Genistein(P<0.05).2. The results of CLSM and Western blot analysis showed that LPA induced the decreased expression of P120ctn in cell membrane,but increased in cytoplasm of SKOV3 cell. After acted by Genistein and LPA, the expression of P120ctn in SKOV3 cell membrane and cytoplasm show an opposite results. However, the total cellular protein of P120ctn of SKOV3 cells detected by RT-PCR was unchanged.The results of Flow cytometry analysis showed that LPA could decreased the proportion of SKOV3 cells in the G1 phase of the cell cycle, increased in the S phase of the cell cycle(P<0.05); Whereas, Genistein combining LPA in culture, the proportion of SKOV3 cells in the S phase was decreased, G2 phase of the cell cycle was rised(P<0.05).The detection of Millicell small chamber invasion showed that LPA could increased the migration and invasion capacity of SKOV3 cell(sP<0.05); Whereas, Genistein combining LPA in culture, cell migration and invasion capacity were decreased(P<0.05).Conclusions:1. The proliferation of SKOV3 cells could be promoted by LPA; but this effect of LPA could be obviously repressed by Genistein.2. LPA could decreased the expression of P120ctn in cell membrane,but increased in cytoplasm of SKOV3 cell. After acted by Genistein and LPA, the expression of P120ctn in SKOV3 cell membrane and cytoplasm show an opposite results. It indicated that Genistein could inhibit the P120ctn expression of SKOV3 cells induced by LPA.3. LPA could decreased the proportion of SKOV3 cells in the G1 phase of the cell cycle, increased in the S phase of the cell cycle. It suggests that LPA promoted the proliferation of SKOV3 cells, but Genistein combining LPA in culture, the proportion of SKOV3 cells in the G2 phase of the cell cycle was rised, S phase was decreased. It suggests that Genistein inhibited SKOV3 cells proliferation.4. The detection of Millicell small chamber invasion showed that LPA could increased the migration and invasion capacity of SKOV3 cells, however, Genistein combining LPA in culture, cell migration and invasion capacity were decreased. This result suggested that Genistein could weaken the migration and invasion capacity of SKOV3 cells.