【作者】 史玉林；

【导师】 张淑兰；

【作者基本信息】 中国医科大学 ， 妇产科， 2005， 博士

【摘要】 目的 检测KLK9和KLK14基因mRNA及其蛋白在正常卵巢、卵巢交界性瘤和卵巢癌组织中的表达差异,并分析KLK9和KLK14基因与卵巢癌临床病理特征的关系及其临床意义。观察雌、孕和雄激素对卵巢癌细胞系中KLK9和KLK14基因mRNA及蛋白表达的影响,以及诱导卵巢癌细胞增殖和凋亡作用。探讨KLK9、KLK14基因和雌、孕及雄激素在卵巢癌发生和侵袭中作用及相关分子机理,为卵巢癌的基因诊断和药物治疗提供科学的实验依据。 方法 一.应用半定量RT-PCR方法,检测55例卵巢癌及其正常对照组织中KLK9和KLK14基因mRNA表达,探讨并分析其与卵巢癌临床病理特征的关系。 二.采用SP免疫组化和/或Western Blot印迹分析法,检测正常卵巢(n=55)、卵巢交界性肿瘤(n=16)、卵巢癌(n=55)组织中KLK9和KLK14蛋白的表达。分析KLK9和KLK14蛋白表达与卵巢癌临床病理特征的关系。 三.雌、孕和雄激素作用SKV卵巢癌细胞系24、48、72小时后,应用Bacton Dickonson FACScan流式细胞仪,测定其细胞周期和细更多还原

【Abstract】 ObjectiveTo test the expression differences of KLK9 and KLK14 gene mRNA and proteins in normal,borderline and malignant ovarian tissues.To analyze the relationship between KLK9 、 KLK14 gene and clinicopathological features of ovarian cancer. To observe the effect of estrogen,progestogen and androgen on the expression of KLK9 and KLK14 gene mRNA in ovarian cancer cell lines; and the effect of induction to the cancer cell proliferation and apoptosis. To explore their roles in tumorgenesis and metastasis of ovarian cancers,to supply an experimental basis for the gene diagnosis and medical treatment of the ovarian cancers.Methods1 、 KLK9 and KLK14 gene mRNA were examined in ovarian cancers (n=55)and the control group (n=55 ) by semiquantitative RT-PCR.2、 KLK9 and KLK14 protein expression were examined in normal ovaries(n=55),borderline ovarian diseases(n=16) and ovarian cancers (n=55) by means of immunohistochemical SP method and Western Blot.3、 We use Bacton Dickonson FACS to test the cell cycles and apoptosis of the SKV cell lines after the use of estrogen、 progestogen andandrogen for 24、 48、 72 hours.4、 We examine the different expression of KLK9 and KLK14gene mRNA and protein in SKV cell lines after the use of estrogen 、 proestogen and androgen by RT-PCR and Western Blot.Results1、 The expression of KLK9 gene mRNA and protein was significantly lower in ovarian cancers than that in normal ovarians (p<0.05 ) .The expression of KLK9 gene mRNA and protein in early stage of ovarian cancer was significantly higher than that in late stage .(p<0.01), but the expression has no relationship with the pathological stages and histological differentiation of ovarian cancers.2、 The expression of KLK14 gene mRNA and protein increased gradually in normal ovaries,borderline tumors and ovarian cancers;and it is higher in borderline and carcinomatous tissues than that in normal ovary tissues (P<0.05). The expression of KLK14 gene mRNA was closely related with the clinicopathological features of ovarian cancers. The experiments showed that the expression of KLK14 gene mRNA and protein was higher in late stage than that in early stage of ovarian cancers, and higher in low differentiated ones than that of in high differentiated cancers. And the expression levels have no relationship with the different histological type.3、 Compared with the control group ,after the induction of estrogen, progestogen and androgen to the ovarian cancer call lines, in estrogen group cells in S phase increased obviously while cells in G2/M phase decreased.(P<0.0l)4、 compared with the control group, the cell lines showed increased expression of KLK9 gene mRNA and protein after the use of estrogen and increased expression of KLK14 gene mRNA and protein after the use of progestogen and increased expression of KLK14gene mRNA and protein after the use of androgen.Conclusions1 、 The down-regulated expression of KLK9 gene mRNA and protein existed in the ovarian cancers. It was correlated with the clinicopathological stage of the ovarian cancer and can be a monitoring marker for the prognosis of ovarian cancer2、 The up-regulated expression of KLK14 gene mRNA and protein existed in the ovarian cancers.It was closely related with the clinicopathological stage and differentiation degree of the cancer,and can be a biomarker to predict the biological behavior.3、 Estrogen,progestogen and androgen are closely related with the progression of the ovarian cancers.Progestogen can enhance the apoptosis of the ovarian cancer cells and inhibit the growth of the cancer cells.The induction of these three hormones to the cancer cells can be a guidance to更多还原