| Since the occult onset, high degree of malignancy, the resistance to chemotherapeutic drugs and other reasons, ovarian cancer has been a serious threat to the lives and health of women. Usually ovarian cancer is found late and sometimes with the pelvic or distant metastasis, which lead to the unsatisfied treatment results and the lower 5-year survival rate. So it is particularly important to explore the mechanism for the development and transfer of ovarian cancer. Transforming growth factorβ(TGF-β) plays a negative role in normal cells growth regulation, but in tumor cells is generally considered to have inhibitory and promoting effects as a double-edged sword. TGF-βplays its biological effects in a very wide range, not only the regulation of cell growth and differentiation, but also the apoptosis, immune suppression, angiogenesis, and close relationship with a variety of oncogenes. TGF-β1 is the focus of our experiment not only because it was found first and now the most studied subtype in TGF-βfamily, but also its wide existence in the human body and as the most important control factor in the occurrence and development of ovarian tumors. Fas (Apo-1/CD95) is a kind of apoptosis-related proteins in the human body, which is related with apoptosis and tumor occurrence, development and metastasis. Fas as an important member of death receptor family, transmit signals into cells through the combination with its receptor FasL, and lead the apoptosis of the cells which express Fas on its surface. In this paper, we will detect the expression of TGF-β1 and Fas in ovarian tumors and to explore its relationship with the development and metastasis of ovarian cancer. Through follow-up investigation of patients we try to explore the relationship of TGF-β1 and Fas with the ovarian cancer recurrence and prognosis.[Materials and methods]Select surgical resection of ovarian tumor tissues from the patient with ovarian cancer from Second Hospital of Jilin University,detect the expression of TGF-β1 and Fas protein and mRNA in ovarian tumor tissues through immunohistochemistry(IHC) and In situ hybridization(ISH), and the expression on the surface by flow cytometry(FCM).[Results]1. The expression of TGF-β1 protein in malignant ovarian cancer was higher than benign ovarian tumors and normal ovarian tissue, while the TGF-β1 mRNA showed higher in malignant ovarian cancer than than benign ones but no difference with the normal ovarian tissue; The expression Fas in malignant ovarian cancer was lower than ovarian tumors and normal ovaries,while no difference between tumers and normal tissue. TGF-β1 and Fas protein positive expression were higher in ascites volume of≥500ml group than that in the ascites volume <500ml group (P <0.05), but no significangt difference between the groups of patient's age, clinical stage, pathological type, histological grade and pelvic lymph node metastasis (P> 0.05); TGF-β1 and Fas proteins in malignant ovarian cancer tissue show significantly difference byχ~2 test (χ~2 = 11.5294, P = 0.0000).2. Through FCM detection, the rate of positive expression of TGF-β1 on malignant ovarian cancer cells surface was higher than ovarian tumor cells (t '= 2.5033, P <0.0500), but showed no significant difference in mean fluorescence intensity (t '=- 1.7005, P> 0.0500).The positive expression rate of Fas protein was not significant different on cell surface of the two kinds of cells(t = 0.8434, P = 0.4235), while the average fluorescence intensity on malignant cells was significantly lower than the expression of ovarian tumors (t = 2.3920, P = 0.0437).3. Through IHC and ISH we observed that in the central gland cavity of cancer nest which is negative there were a large number of necrotic tissue with scattered cancer cells with TGF-β1 protein and mRNA strong positive expression. This phenomenon is considered the negative growth regulation effects of TGF-β1 in the ovarian cancer. In some cases we can see the existence of necrotic tissue with a large number of cancer cells in agglomeran surrounding cancer nests showed TGF-β1 strong positive expression, with the sharp contrast to negative expression of cancer nests the tumor, and this situation may suggest TGF-β1 promotes cancer cells stromal invasion. The two phenomena are not seen in Fas protein or mRNA expression.4. In IHC and ISH of ovarian cancer, we can see there are a large number of nucleated cells in vascular especially neutrophils and mononuclear cells, which were strongly positive TGF-β1 expression. This may suggests relationship between TGF-β1 and tumor angiogenesis and invasion. Similer phenomenon was also found in Fas IHC and ISH, which may suggest the attack to the cancer cells.5. According to the situation of patients recurrence make Kaplan-Meier curve and Log-Rank test proved lower relapse rate in TGF-β1 protein positive patients than the TGF-β1-negative ones(P <0.05), Fas expression and relapse rate showed no correlation (P > 0.05).[Conclusions and innovation]Through the comparison of IHC and ISH results we consider that TGF-β1 in ovarian cancer cells may be the results of abnormal expression of gene expression; While the abnormal expression of Fas protein level does not involve genetic abnormalities, but may be caused by other reasons such as Fas protein structure and function changes. Combined analysis of two methods is the first innovation of this article. Also in this experiment by flow cytometry we detected TGF-β1 and Fas protein on ovarian tumor cell surface, both of which play their roles in transmission of signals through a combination with cell surface receptors, and found consistent with the results of IHC. This experiment using flow cytometry, compared with the traditional immunohistochemical methods, which may more objectively reflect cytokines in ovarian tumor cell surface, and avoid subjective factors interference on the experimental results. In addition, the experimental procedures and steps of low cytometry are more simplified, which can avoid some injury antigens caused bu some procedures, such as the fixation of the tissue. So it enhanced the authenticity and reliability of the experimental results which is the second innovation. By a short follow-up of patients we make a relapse curve to analysis the situation of recent recurrence with TGF-β1 and Fas. It is only a reference, and we still need to do a lot of work.In IHC and ISH we observed in the center of nest cavity there were a large quantities of necrotic tissue with the cancer cells strong positive expression of TGF-β1, which have not been reported previously as the third innovation of our paper. This may be a new evidence that suggest TGF-β1 play a role in promoting tumor cells invasion and metastasis into adjacent stromal tissue. Forth, we found there are lots of nucleated cells in the vascular of ovarian cancer tissue, in which TGF-β1 and Fas express strong positively may also mean that the two factors play roles in tumor through a leukocyte-related mechanism. Of course, only according to the expression of strong positive in immunohistochemistry, we can only make speculate reasonably. To explore effect of TGF-β1 in the ovarian cancer metastasis still need more deep work.
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