| Some ovarian high grade serous adenocarcinoma (HGSC) may have originated in the fallopian tube fimbria epithelial cells (mainly oviduct epithelial secretory cells). Under the environment with a variety of factors such as the stimulation, "P53 signature" appeared, followed by tubal intraepithelial carcinoma (serous tubal intraepithelial carcinoma, STIC), and finally became invasive serous adenocarcinoma [4,5].Epithelial-mesenchymal transitions (EMT) refers to the morphology of epithelial cells occuring fibroblasts and mesenchymal’s phenotype change and obtaining the ability to migrate. It was correlate with regeneration of tissue reconstruction, tumor development and metastasis. Studies have shown that normal epithelial cells can generate EMT under the action of various factors, participating in tumorigenicity [3]. High mobility group proteinA2 (HMGA2) is a member of HMGA family and it was correlate with the expression level of the degree of malignancy, metastasis and prognosis.Currently, many studies have focused on what happened after "P53 signature" appears, especially STIC. but there were few studies for the change of cells before "P53 signature". However, and studies of events that occur at this stage can conduce on the pathogenesis, early diagnosis and targeted intervention of HGSC.For the purpose of above, the study explored whether the FTE cells occurred EMT after the FSH stiumulation, and the effcet of HMGA2 in this process.Part I Biomarker expression in normal fimbriae:Comparison ofhigh- and low-grade serous ovarian carcinomaObject:The objective of this study was to assess the difference in fimbriae of high-and low-grade ovarian serous carcinoma (OSC).Methods:The fimbriae of normal appearance [without serous tubal intraepithelial carcinoma (STIC)] from 28 patients with high-grade OSCs and 24 patients with low-grade OSCs were assessed for the expression of 6 markers [E-cadherin, matrix metalloproteinase-2 (MMP-2), phospho-AKT (pAKT), cyclooxygenase-2 (COX-2), vascular endothelial growth factor (VEGF) and p53] using immunohistochemistry. Sectioning and extensively examining the fimbria (SEE-FIM) was performed to exclude fimbrial involvement for all the cases.Results:The immunostaining levels of pAKT and COX-2 were significantly higher in the fimbriae of normal appearance from high-grade OSCs compared with low-grade OSCs (61vs.8% and 71 vs.21%; P=0.005 and 0.007, respectively). The immu nostaining of E-cadherin was significantly higher in the fimbriae of low-grade OSCs compared with high-grade OSCs (83 vs.21%; P=0.003). The remaining 3 markers (MMP-2, VEGF and p53) had similar expression in low-and high-grade OSCs (21 vs.13%; 25 vs.21%; and 14 vs.8%; P=0.78,0.86 and 0.82, respectively).Conclusion:Our results suggest marked biological differences in the behavior of the fimbriae in high- and low-grade OSCs and indicate that proliferation, cell adhesion and the inflammatory microenvironment of fimbriae in high-grade OSCs without STIC had changed prior to p53 mutation.Part Ⅱ FSH promoted the occurrence of EMT in FTE cells of HGSCObjective:To understand whether FSH can promote EMT by simulating FTE of HGSC.Methods:Based on primary culture of FTE cell without tumor involving and P53(-) of HGSC and LGSC, FSH stimulated the FTE cells with different concentrations (0,10,20,40,80,160 mIU/mL) and at different times (respectively 0,12,24,48,72 h). Cell morphology changes were observated and the protein expression of EMT markers E-Cadherin, N-Cadherin, vimentin, MMP-2, P53 were detected by Western-blot, and the mRNA expression were detected by RT-PCR.Results:After 48h with FSH with concentrations 40mIU/ml acting on FTE cells, the appearance of epithelial cells evolved into spindle fiber morphology. EMT markers, E-Cadherin was significantly reduced, while the N-Cadherin, vimentin, MMP-2 significantly increased, which confirmed the occurrence of EMT. These changes were not seen in the FTE cells of LGSC.Conclusion:FSH promoted the occurrence of EMT in FTE cells of HGSC.Part Ⅲ FSH promoted the occurrence of EMT in FTE cells of HGSC by regulating HMGA2Object:The objective of this study was to assess the role of HMGA2 in the occurrence of EMT in FTE cells of HGSC by FSH stimulating.Methods:Based on primary culture of FTE cell without tumor involving and P53(-) of HGSC, FSH stimulated the FTE cells with different concentrations (0,10,20,40,80 mIU/mL) and at different times (respectively 0,12,24,48,72 h).The protein expression of HMGA2 were detected by Western-blot, and the mRNA expression of let-7b were detected by RT-PCR.Transfected with HMGA2 shRNA, the FTE cells were stimulated by FSH. EMT and the protein expression of EMT markers E-Cadherin, N-Cadherin, vimentin, MMP-2 were observed. The expression of HMGA2 was detected by knockdown of let-7b.Results:Treatment with FSH signifi cantly increased HMGA2 expression in a time-dependent manner and the let-7 expression levels decreased gradually over time in the FTE cells of HGSCs. Transfected with HMGA2 shRNA, the FTE cells did not occur EMT by treatment with FSH. And the changes of expression of EMT markers, E-Cadherin, N-Cadherin, vimentin, MMP-2 were not detected. Knockdown of let-7b stimulated HMGA2 expression.Conclusion:FSH promoted the occurrence of EMT in FTE cells of HGSC by regulating HMGA2. Let-7b regulated HMGA2 expression and FSH stimulates HMGA2 expression by down regulating let-7b.In summary, FSH promoted the occurrence of EMT in FTE cells of HGSC, and the optimum conditions were concentration 40mIU/ml and time 48h. FSH can downregulated the expression of let-7b and upregulated the expression of HMGA2 by it. expression. FSH promoted the occurrence of EMT in FTE cells of HGSC by regulating HMGA2 and the phenomenon were not found in LGSC. The findings suggest FSH promoted the occurrence of EMT in FTE cells of HGSC by regulating HMGA2 and explored associated molecular regulatory mechanisms, It’s helpful for the new theoretical basis and therapeutic targets for the occurrence, early diagnosis and treatment of HGSC. |
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