【作者】 王红霞；

【导师】 李国利； 陈钢；

【作者基本信息】 扬州大学 ， 病理学与病理生理学， 2009， 硕士

【摘要】 目的:通过检测bcl-2、bax、survivin、smac和fas在卵巢黏液性肿瘤中的表达,分析卵巢黏液性肿瘤发生发展过程中凋亡调控相关基因的作用,进而揭示凋亡调控机制的紊乱是卵巢黏液性肿瘤恶变过程中的一个重要环节。方法:应用免疫组化SABC法,检测bcl-2、bax、survivin、smac和fas在55例原发性卵巢黏液性肿瘤中的表达;应用免疫电镜胶体金标记法检测smac在原发性卵巢黏液性肿瘤中的亚细胞表达部位及表达趋势。结果:(1)Bcl-2在良性、交界性和恶性组中的阳性表达率分别为10%、46.67%和85%;良性与交界性、交界性与恶性比较均具有统计学差异(P<0.05),良性与恶性比较有显著统计学差异(P<0.01)。三组中bax阳性表达率分别为80%、60%和10%;良性与交界性比较无统计学差异(P>0.05),交界性与恶性、良性与恶性比较有显著统计学差异(P<0.01)。Bcl-2/bax比率在良性、交界性和恶性组平均值分别为0.24±0.42、0.85±1.56、15.46±19.58;良性与交界性比较无统计学差异(P>0.05);而交界性与恶性、良性与恶性比较均有统计学差异(P<0.05)。(2)Survivin在良性、交界性和恶性组中的胞浆阳性表达率分别为10%、80%和100%;良性与交界性、良性与恶性比较均有显著统计学差异(P<0.01),而交界性与恶性比较无统计学差异(P>0.05)。Survivin胞核阳性表达率分别为5%、40%、25%;良性与交界性组比较有统计学差异(P<0.05),良性与恶性以及交界与恶性比较均无统计学差异(P>0.05)。三组中smac阳性表达率分别为95%、60%和15%;良性与交界性、交界性与恶性比较均有统计学差异(P<0.05) ,良性与恶性比较有显著统计学差异(P<0.01)。(3) Fas在良性、交界性和恶性组中的阳性表达率分别为65%、53.33%和45%;良性与交界性、交界性与恶性、良性与恶性比较均无统计学差异(P>0.05)。(4) Bcl-2、bax、survivin、smac基因间存在不同程度的相关性。Bcl-2和survivin呈正相关(r=0.870, P<0.01);bax和smac呈正相关(r=0.944, P<0.01);bcl-2和bax呈负相关(r=-0.208, P<0.01);survivin和smac呈负相关(r= -0.153, P<0.01);bcl-2和smac呈负相关(r= -0.146,P<0.01);bax和survivin呈负相关(r= -0.185,P<0.01)。(5) Smac金标颗粒在良性、交界性和恶性组中计数分别如下:其在胞浆内分别为28.79±6.68、13.91±1.98和6.98±0.96;良性与交界性以及良性与恶性比较有显著统计学差异(P<0.01),交界与恶性组间比较有统计学差异(P<0.05);线粒体内计数分别为24.11±7.18、11.06±1.87和5.24±1.72,良性与交界组以及良性与恶性组间比较有显著统计学差异(P<0.01),交界与恶性组间比较有统计学差异(P<0.05);线粒体外分别为4.67±2.98、2.86±0.95和1.74±1.25,组间比较均无统计学差异(P>0.05)。结论:1、在卵巢粘液性肿瘤的良恶性演变过程中存在凋亡调控的紊乱,其中线粒体途径可能为诱导凋亡紊乱的主要通路。Survivin的表达调节是一早期事件,但其表达部位的改变则是一连续性过程,survivin的胞核表达可能与肿瘤的良性生物学行为有关;而bax则是一晚期事件,交界性卵巢粘液性囊腺瘤患者检测到bax的缺如提示有潜在恶变的可能;bcl-2和smac的表达异常在卵巢粘液性肿瘤的良恶性演变过程中是一连续性事件。2、Bcl-2/bax比率可能与卵巢粘液性肿瘤的良恶性程度密切相关。3、在卵巢粘液性肿瘤的恶性演变过程中,下调的smac蛋白主要为储存在线粒体膜间的非活性形式蛋白,而非释放到胞浆中的活性形式蛋白。更多还原

【Abstract】 Objective: To analysis the function of apoptosis regulation related genes during the development and progress of ovarian mucinous tumors, by detecting the expressions of bcl-2、bax、survivin、smac and fas in ovarian mucinous tumors,so as to reveal the dysfunction of apoptosis regulation mechanism is an important link during the canceration of ovarian mucinous tumors.Methods: 55 paraffin specimen of primary ovarian mucinous tumors were collected. IHC were used to detect the proteins of bcl-2、bax、survivin、smac and fas;the immune electron microscope marked by colloidal gold were used to detect the deuto-cell expression position and the trend of smac protein .Results: (1)The positive expression rates of bcl-2 in benign、borderline and malignant ovarian mucinous tumors were 10%、46.67% and 85%; and there were different significances of bcl-2 between benign and boderline(P<0.05),boderline and malignant(P<0.05),benign and malignant(P<0.01).Bax were 80%、60% and 10%;and there were different significances of bax between boderline and malignant(P<0.01), benign and malignant(P<0.01),while no significance between benign and borderline (P>0.05).The ratios of bcl-2/bax were 0.24±0.42、0.85±1.56、15.46±19.58 respectively in benign、borderline and malignant ovarian mucinous tumors;and there were different significances between boderline and malignant(P<0.05), benign and malignant (P<0.05),while no significance between benign and borderline(P>0.05).(2)The cytoplasm positive expression rates of survivin in benign、borderline and malignant ovarian mucinous tumors were 10%、80% and 100%; and there were different significances of survivin between benign and boderline(P<0.01), benign and malignant(P<0.01),while no significance between boderline and malignant (P>0.05). The nucleus positive expression rates of survivin in benign、borderline and malignant ovarian mucinous tumors were5%、40%、25%;and there was different significance of survivin between benign and boderline(P<0.05),while no significance between boderline and malignant, benign and malignant(P>0.05).Smac were 95%、60% and 15%;and there were different significances of smac between benign and boderline (P<0.05),boderline and malignant(P<0.05), benign and malignant(P<0.01).(3) The positive expression rates of fas in benign, borderline and malignant ovarian mucinous tumors were 65%、53.33% and 45%;and there were no different significance of fas among benign、borderline and malignant groups(P>0.05).(4) There were correlations among bcl-2、bax、survivin and smac proteins. There was postive correlation between bax and smac(r=0.944,P<0.01);bcl-2 and survivin(r=0.870,P<0.01).There was negative correlation between bcl-2 and bax (r=-0.208,P<0.01);survivin and smac(r=-0.153,P<0.01);bcl-2 and smac(r=-0.146, P<0.01);bax and survivin(r= -0.185,P<0.01).(5) The smac granules marked by colloidal counted in the three groups were as follows: the intracytoplasm smac granules were 28.79±6.68、13.91±1.98 and 6.98±0.96;and there were different significances between benign and boderline (P<0.01),boderline and malignant(P<0.05),benign and malignant(P<0.01).The smac granules storing between the two layer membrane of mitochondrion were 24.11±7.18、11.06±1.87、5.24±1.72 respectively;and there were different significances between benign and boderline(P<0.01),boderline and malignant(P<0.05),benign and malignant (P<0.01).The smac granules located out of mitochondrial membrane were 4.67±2.98、2.86±0.95 and 1.74±1.25;and there were no different significances among them(P>0.05).Conclusions: (1) The dysfunction of apoptosis regulation exists in ovarian mucinous tumors during the progress from benign to malignant,and the mitochondrion path mignt be the major pathway inducing dysfunction of apoptosis.The regulation of the expression of survivin is a pristine event while the change of the expression position is a continuous event ,and the nucleus expression is positive correlation with the benign biological behaviors;however,bax is a late event ,detecting the absence of bax protein in borderline ovarian mucinous tumor patient maybe suggest a potential canceration;the abnormal expressions of bcl-2 and smac protein are a continuous event during the progress from benign、borderline to maligant groups (2)The ratio of bcl-2/bax may be more related with the benign or malignant extents in ovarian mucinous tumors (3)The down regulated smac proteins mainly are the inactive patterns storing between the two layer membrane of mitochondrion,nor the releasing active patterns out of mitochondrial membrane during the progress from benign to malignant ovarian mucinous tumors.更多还原