| Objective: The cytotrophoblastic cells (pseudomalignancy) and malignanttumor cells have the similarities in many biological processes including for instance cell proliferation, differentiation and remodelation of vascularisation, immune escape, cell migration and apoptosis. So the crossing and confluence of methodology berween the cytotrophoblastic cells and malignant tumor cells will offer new idea for elucidating the intrinsic mechanisms and clinical therapy for the two complicated vital phenomena.So the model of mouse blastocysts co-cultured with ovarian carcinoma cells was established to explore dynamic interaction and difference in the invasive potential and its related gene expression between the two kinds of life cells under the same microenviroment in vitro. Its provided a research platform to explain the mechanisms of cell interaction and a crossing study between embryo implantation and invasion of tumor. To approach the effect of the uterine deciduas in peri-implantation on ovarian cancer cells invasion and metastasis, and its mechanisms. The trophoblastic cells orderly invading maternal endometrium in vivo was explored to elucidate the molecular mechanisms and therapeutic targets for malignant tumor cells form embryo implantation angle as an anti-tissue invasion model.Methods: The first part, the model of mouse blastocysts co-cultured withhighly metastatic ovarian carcinoma cell HO-8910 PM was established to observe the interaction and whole biological behaviors changes of the two kinds of life cells under the same microenviroment in vitro;MTT assay was used to examine changes of HO-8910 PM cell adhesion;transwell chamber assay was performed to determineits effect on invasion and migration capacities of HO-8910 PM;and mRNA levels of MMP-2, MMP-9, TIMP-1, TIMP-2 were assessed by RT-PCR. The second part, the crude extract from mice uterine deciduas during peri-implantation(dO^ d4~d8N dl3) was added to the medium of HO-8910 PM, then MTT assay was used to examine the growth and adhesive rate of those experimental and control groups;flow cytometry was performed to determine the cell life cycle and apoptosis rate of each group;transwell chamber assay was performed to examine its effect on invasion and migration capacities of every group;protein levels of MMP-2, MMP-9, TIMP-1, TIMP-2 were assessed by gelatin zymography and reverse-zymography;and mRNA levels of MMP-2, MMP-9, TIMP-1, TIMP-2 were assessed by RT-PCR.Results:1. After 24h of being co-cultured, mouse blastocyst escaped from zone pellucida and hatched on the underlying malignant ovarian tumor cells,then the partially hatched blastocysts attached to the co-culture cancer cells. The blastocysts inserted the gaps among tumor cells in finger shape, gradually push underlying highly proliferative and divisive tumor cells and then freely invaded them. However, the ovarian tumor cells only accumulated on the periphery of trophoblast cells as rotundity or ellipse shape.2.MTT assay revealed: compared with control group, the adhesive rate of co-culture group was significantly decreased. After co-culture with mouse blastocysts, number of HO-8910 PM cells to penetrate polycarbonates coated with or without Matrigel was significantly lower (P <0.01).3. After co-culture with mouse blastocysts, HO-8910 PM mRNA levels of MMP-2, MMP-9 were significantly lower, on the contrary, mRNA levels of TIMP-1, TIMP-2 were increased (no significance), ratios of MMP/TIMP were significantly lower.4. The crude extract from mice uterine deciduas (dO> d4~d8^ dl3) was addedto the medium of HO-8910 PM, the growth rates of d4, d5, d6 were lower than the rate of dO at 24, 48h (P <0.05);the growth rates of d4, d5, d6 were significantly lower than control group at 72h. Compared control group, the adhesive rates of d5 group were increased at 24,48h, and decreased at 72h, but all was lower than dO group.5. All kinds of proteins extracted from mouse uterine deciduas could up-regulate the percentage of G0/G1 phase and apoptosis rate at 48h (compared to control group 59.65%;0.38%), especially d5 group.6. Control group and experimental groups could penetrate polycarbonates coated with or without Matrigel , and number of experimental groups was significantly lower,especially d5 group. Followed co-culture time lengthening, the number decreased.7. When added all kinds of protein, the expression of MMP-2, MMP-9 mRNA of d4,d5,d6 reduced significantly at 24h,48h,72h, especially d5 group, at the same time, TIMP-1, TMP-2 mRNA of d5 group were up-regulated.8. Gelatin zymography and reverse-zymography showed: proteins of MMP-2, MMP-9 were secreted decreased significantly by d4,d5,d6 groups, especially d5 group, compared to dO group;meanwhile, TIMP-1, TIMP-2 of d5,d6 group were increased.Conclusion:1. When the model of mouse blastocysts co-cultured with highly metastatic ovarian carcinoma cell HO-8910 PM was established, trophoblast giant cells differentiated from blastocysts became aggressive "malignant cells", however, tumor cells lost their characteristic invasion.2. After being co-cultured, mouse blastocyst can make invasiveness of HO-8910 PM cells decrease,and change invasion-related adhesive and mobile capacity in vitro;blastocyst can also down-regulate the expression of MMP-2 ^ MMP-9 mRNA ofHO-8910 PM, depressed the ratio of MMP/TIMP.3.Crude extract from mice uterine deciduas pregnancy 5 days can make invasiveness of HO-8910 PM cells decrease,and change invasion-related adhesive and mobile capacity in vitro.4. Crude extract from mice uterine deciduas pregnancy 5 days can significantly lower the expression of MMP-2, MMP-9 protein and iriRNA, at the same time, up-regulate the expression of TIMP-1, TIMP-2.5.Our study revealed, mouse blastocyst and endometria during implantation stage could inhibit cancer metastasis, possessed the potential to be a kind of anticancer, but we need to investigate its mechanism of action.
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