【作者】 赵红兵；

【导师】 李惠翔； 吕新全；

【作者基本信息】 郑州大学 ， 病理学与病理生理学， 2008， 硕士

【摘要】 背景与目的:在全球性女性发生的恶性肿瘤中,乳腺癌是最常见的恶性肿瘤,居女性恶性肿瘤的首位。近年来乳腺癌发病率呈上升趋势,发病年龄日趋年轻化。目前,临床上首次就诊的乳腺癌病人中,因早期诊断、判断预后的生物指标缺乏,大多数为中、晚期。检测乳腺癌标志物可反映肿瘤的生物学特性,对辅助诊断、指导治疗、判断预后均有重要意义。乳腺癌的发生、发展是一个多阶段多步骤的复杂过程,抗凋亡是乳腺癌癌细胞实现侵袭、转移、增殖的关键步骤之一。Survivin作为迄今发现最强的凋亡抑制因子,其组织分布有明显的细胞选择性,主要表达于胚胎、发育的胎儿组织和大多数肿瘤组织内。Survivin具有抑制细胞凋亡、促进细胞转化并且参与细胞的有丝分裂、血管的生成和肿瘤细胞耐药性的产生等作用。Bcl-2作为凋亡抑制因子之一,通过高表达所介导的细胞凋亡障碍,使遗传物质的突变率增加,从而参与多种肿瘤的发生过程。PCNA作为细胞增殖因子之一,与细胞周期中的DNA合成期(S期)有密切相关性,其表达可作为乳腺癌的一种辅助诊断指标。本研究拟采用免疫组织化学方法(S-P法)检测Survivin、Bcl-2和PCNA在乳腺癌组织中的变化特征,探讨Survivin、Bcl-2和PCNA三者与乳腺癌的发生、发展和预后的相关性,增加对乳腺癌多阶段演进的分子机理的了解,为临床乳腺癌生物防治提供重要的理论基础和手段。材料和方法:60例乳腺癌患者标本来自河南省郑州大学第一附属医院2006年3月至2007年1月手术切除标本,均为女性,年龄33～75岁,中位年龄51岁,患者术前均未接受化疗、放疗及其它治疗。所有标本经组织病理学检查证实为乳腺癌浸润性导管癌。所有手术切除标本均经10%甲醛固定、石蜡包埋、连续切片,进行组织病理学和免疫组化检查。免疫组化采用S-P法检测乳腺癌组织中Survivin、Bcl-2和PCNA蛋白的表达。采用SPSS11.5统计软件处理,各组间率的比较采用x~2检验,α=0.05作为检验标准。结果:1.Survivin表达主要位于细胞浆,阳性标志为乳腺癌癌细胞的胞浆中出现棕黄色颗粒,Survivin阳性表达率为70.00%(42/60)。按组织学分级Survivin蛋白在乳腺癌组织中免疫反应阳性率分别为:Ⅰ+Ⅱ级61.90%(26/42)、Ⅲ级38.10%(16/42),各组之间无显著性差异(P＞0.05)。Survivin蛋白在≤45岁组的阳性表达率为69.23%(18/26);在＞45岁组的阳性表达率为70.59%(24/34);在不同年龄组患者中Survivin蛋白的阳性表达无显著性差异(P＞0.05)。Survivin蛋白在发生局部淋巴结转移组的阳性表达率为89.29%(25/28);明显高于未发生局部淋巴结转移组,二者有显著性差异(P＜0.01)。Survivin蛋白在ER、PR阳性中的阳性表达率分别为:61.11%(22/36);76.92%(20/26)。Survivin蛋白免疫表达在ER、PR阴、阳性中无显著差异(P＞0.05)。2.Bcl-2表达定位于胞浆或胞膜,阳性标志为胞浆或胞膜出现棕黄色颗粒,Bcl-2阳性表达率为66.67%(40/60)。按组织学分级Bcl-2蛋白乳腺癌免疫反应阳性的细胞阳性率分别为:Ⅰ+Ⅱ级30.00%(12/40)、Ⅲ级70.00%(28/40),两组之间有显著差异(P＜0.01)。Bcl-2蛋白在≤45岁组的阳性表达率为65.38%(17/26);在＞45岁组的阳性表达率为67.65%(23/34)。乳腺癌组织Bcl-2蛋白在不同年龄患者中的阳性表达差异无显著性(P＞0.05)。Bcl-2蛋白在发生局部淋巴结转移组的阳性表达率为82.14%(23/28);明显高于未发生局部淋巴结转移组,二者有显著性差异(P＜0.05)。Bcl-2蛋白在ER、PR阳性中的阳性表达率分别为:61.11%(22/36);73.08%(19/26)。Bcl-2蛋白免疫表达在ER、PR中无显著差异(P＞0.05)。3.在乳腺癌组织中Survivin蛋白的表达与Bcl-2蛋白的表达具有显著的相关性(P＜0.01)。PCNA阳性表达主要位于肿瘤细胞核,胞浆可见弱阳性,阳性表达率为71.67%(43/60)。Survivin蛋白表达阳性的乳腺癌PCNA阳性明显高于Survivin蛋白表达阴性的乳腺癌细胞PCNA,二者之间差异有统计学意义(P＜0.01)。Bcl-2蛋白表达阳性的乳腺癌PCNA阳性明显高于Bcl-2蛋白表达阴性的乳腺癌细胞PCNA,二者之间差异有统计学意义(P＜0.01)。结论:1.Survivin、Bcl-2蛋白在乳腺癌组织中的阳性表达率高,表明二者参与了乳腺癌的发生、发展过程。二者在有淋巴结转移的乳腺癌组织中的阳性表达率高于无淋巴结转移的乳腺癌组织,提示Survivin和Bcl-2可能是判断乳腺癌预后的重要标志物。二者在乳腺癌组织中的表达有明显相关性,提示Survivin与Bcl-2的共同表达可能在乳腺癌组织的发生、发展、浸润、转移中发挥着重要的协同作用。2.联合检测Survivin、Bcl-2与PCNA在乳腺癌组织中的表达可能在判断乳腺癌预后方面起重要作用。更多还原

【Abstract】 Background and Purpose:In the global female occurs in the malignant tumor, breast cancer is the most common malignant tumor,occupies the feminine malignant tumor the first place.In recent years breast cancer disease incidence rate assumed the trend of escalation,the morbidity age is youth oriented day by day.At present,on clinical receives a medical examination for the first time in breast cancer patient, because the early diagnosis,the judgment prognosis’s biological indicator lacks, majority for intermediate stage,later period.Breast cancer designated object examination may reflect that the tumor the biology characteristic,to the auxiliary diagnosis,the instruction treatment,the judgment prognosis has the important meaning.Breast cancer’s occurrence of,the development are more than stage multi-step complex processes,anti-perishes is weakly breast cancer cancer cell realizes one of the attack,the shift,multiplication committed steps.Survivin took discovered until now strongest perished weakly the inhibiting factor,its organization distribution had the obvious cell selectivity,main expression in embryo,in growth embryo organization and majority tumor organizations.Survivin has the suppressor cell to perish weakly,the promotion cell transformation and the participation cell’s mitosis,blood vessel’s production and functions and so on tumour cell drug resistance production.bcl-2 takes perishes weakly one of inhibiting factors,lies between the cell which through the high expression leads to perish weakly the barrier,makes the genetic material the rate of mutation to increase,thus participates in many kinds of tumors the occurrence processes.PcNA one of as cell multiplication factors,(the S time) has the close relevance with mitotic cycle’s in DNA synthesis time,its expression may take breast cancer’s one kind of auxiliary diagnosis target.This research plans to use the immunity histochemical method(the S-P law) to examine Survivin,Bcl-2 and PCNA,in breast cancer organizes the change characteristic.Discusses Survivin,Bcl-2 and PCNA three and breast cancer’s occurrence,the development and the prognosis relevance.Increases to breast cancer multi-phased evolution molecular mechanism’s understanding.Provides the important rationale and the method for the clinical breast cancer biological controls.Materials and Methods:60 example breast cancer patient specimen surgery excises the specimen from the Henan Province Zhengzhou University first affiliated hospital March,2006 to January,2007.all Is the female,the age 33～75 years old.Before the patient technique,has not accepted the chemotherapy,the radiotherapy and other treatments.All specimens after organization pathology inspection confirmation for breast cancer wetting quality drive pipe cancer.All surgeries excise the specimen to pass through 85%alcohol fixation,the paraffin wax embedding,the serial section,carries on the organization pathology and the immunity group inspection.The immunity group uses the S-P law examination breast cancer to organize Survivin,bcl-2 and the PCNA protein expression.Uses SPSS11.5 statistics software processing,each group of rate comparisons use x~2 the examination,the inspection standards takeα=0.05.Results:1.The Survivin expression is mainly located at the cytoplasma,the masculine gender symbolized that presents the yellowish brown pellet for in breast cancer cancer cell’s cytoblastema,the Survivin masculine gender expression rate is 70.00%(42/60). Grades the survivin protein according to the histology,in breast cancer organizes the immune response masculine gender rate respectively is:Ⅰ+ⅡLevel 61.90% (26/42),ⅢLevel 38.10%(16/42),each group of between non-significance difference (P＞0.05).The survivin protein in≤45 year-old group’s masculine expression rate is 69.23(18/26);In the＞45 year-old group’s masculine expression rate is 70.59 (24/34).Breast cancer organizes the survivin protein not to have the significance in disparity in age patient masculine expression difference(P＞0.05) the survivin protein is having the partial lymph node shift group’s masculine expression rate is 89.29% (25/28);Is higher than obviously has not had the partial lymph node shift group,the two have the significance difference(P＜0.01).The Survivin protein in ER,in the PR masculine gender’s masculine expression rate respectively is:61.11%(22/36);76.92% (20/26).The Survivin protein immunity expression in ER,PR are cloudy,in masculine gender not remarkable difference(P＞0.05).2.The Bcl-2 expression locates in the cytoblastema either the amnion,the masculine gender symbolized that presents the yellowish brown pellet for the cytoblastema or the amnion(picture 6.7),the Bcl-2 masculine gender expression rate is 66.67% (40/60).Grades the Bcl-2 protein according to the histology,in breast cancer organizes the immune response masculine gender rate respectively is:Ⅰ+ⅡLevel 30.00%(12/40),ⅢThe level 70.00%(28/40),between two groups has the remarkable difference(P＜0.01).The Bcl-2 protein in≤45 year-old group’s masculine expression rate is 65.38(17/26);In the＞45 year-old group’s masculine expression rate is 67.65 (23/34).Breast cancer organizes the Bcl-2 protein not to have the significance in disparity in age patient masculine expression difference(P＞0.05).The Bcl-2 protein is having the partial lymph node shift group’s masculine expression rate is 82.14% (23/28);Is higher than obviously has not had the partial lymph node shift group,the two have the significance difference(P＜0.05).The Bcl-2 protein in ER,in the PR masculine gender’s masculine expression rate respectively is:61.11%(22/36);73.08% (19/26).Bcl-2 protein immunity expression in ER,PR not remarkable difference (P＞0.05).3.In breast cancer organizes the Survivin protein expression and the Bcl-2 protein expression has the remarkable relevance(P＜0.01).The PCNA masculine gender expression is mainly located at the tumor cell nucleus,the cytoblastema obviously weak masculine gender,the masculine expression rate is 71.67%(43/60).Survivin the protein expression masculine breast cancer PCNA masculine gender is higher than Survivin obviously protein expression negative mammary gland cancer cell PCNA, between the two difference has statistics significance(P＜0.01).Bcl-2 the protein expression masculine breast cancer PCNA masculine gender is higher than Bcl-2 obviously protein expression negative mammary gland cancer cell PCNA,between the two difference has statistics significance(P＜0.01).Conclusions:1.Survivin,Bcl-2 protein,in breast cancer organizes the masculine expression rate to be high,indicated that the two participated in breast cancer’s occurrence,the developing process.The two in have breast cancer which the lymph node shifts to organize the masculine expression rate to be higher than the non-lymph node shift breast cancer organization,prompts Survivin and Bcl-2 possibly is judges breast cancer prognosis the important designated object.The two in breast cancer organizes the expression to have the obvious relevance,prompts Survivin and the Bcl-2 common expression in the occurrence which,the development,the infiltration,the shift organizes at breast cancer is possibly playing the important synergism.2.Examines Survivin,Bcl-2 and PCNA jointly,in breast cancer organizes the expression possibly to play the influential role in the judgment breast cancer prognosis aspect.更多还原