BAD、p-BAD及p-AKT蛋白在乳腺癌癌变过程中的表达及意义

用免疫组织化学S-P法检测131例乳腺石蜡包埋组织、Western Blotting法检测27例乳腺浸润性导管癌及癌旁乳腺组织中Bcl-xl/Bcl-2相关死亡启动子(BAD)、磷酸化Bcl-xl/Bcl-2相关死亡启动子(p-BAD)、磷酸化丝氨酸/苏氨酸蛋白激酶B(p-AKT)的表达情况,分析三种蛋白在乳腺癌组织中表达的意义及其与临床病理特征间的关系。结果免疫组化结果显示在乳腺癌癌变过程中三种蛋白表达阳性率呈递增趋势,三种蛋白在乳腺普通导管增生和重度不典型增生及原位癌间阳性表达率差异有统计学意义(P值分别为0.022、0.023、0.001);p-BAD蛋白在重度不典型增生及原位癌组与浸润性导管癌组间差异有明显统计学意义(P=0.004)。三种蛋白与乳腺浸润性导管癌患者的年龄、临床分期、肿瘤大小无明显关系,但均与组织学分级相关(P值分别为0.026、0.038、0.026)。p-AKT蛋白表达与腋窝淋巴结转移有关(P值为0.016)。Western Blotting结果显示乳腺癌组织中三种蛋白含量明显高于癌旁组织(P值分别为0.002、0.002、0.003)。在乳腺癌组织中BAD与p-BAD及p-BAD与p-AKT蛋白表达量呈正相关性(P值分别为0.011、0.032)。认为这三种蛋白表达水平可提示乳腺导管非典型增生向乳腺癌的转化潜能;PI3-K/Akt传导路径在乳腺癌癌变过程中具有重要作用。     

ERK信号蛋白在乳腺导管上皮癌变过程中的表达

目的 检测细胞外信号调节激酶（ERK）信号蛋白在人乳腺癌和乳腺增生症组织中的表达,探讨ERK在乳腺导管上皮细胞癌变过程中的作用.方法 采用免疫组织化学S-P法,检测67例乳腺组织中ERK信号蛋白的表达.结果 ERK信号蛋白在乳腺导管单纯性增生组织中呈弱表达,阳性率为25%（2/8）,35例乳腺导管不典型增生中26例ERK阳性表达,阳性率为74.3%（P＜0.05）.24例乳腺癌组织中22例ERK阳性表达,阳性率为91.6%,与增生组比较有显著差异（P＜0.05）.结论 ERK信号蛋白在乳腺癌的组织发生中起关键作用,其表达强度与乳腺导管上皮的恶性转化有关.     

浸润性乳腺癌组织中14-3-3σ、p73α蛋白表达及临床意义

目的探讨表皮细胞特异性标志物14-3-3σ、p73α蛋白在浸润性乳腺癌发生发展中的作用。方法选择我院手术切除的浸润性乳腺癌组织标本94份（浸润癌组）,导管原位癌15份（原位癌组）,上皮不典型增生15份（不典型增生组）,导管内乳头状瘤15份（乳头状瘤组）,乳腺增生症20份（增生症组）,乳腺癌旁5 cm组织10份（癌旁组）,采用免疫组化PV-9000两步法检测各组14-3-3σ、p73α蛋白表达情况,并分析两者间及与浸润性乳腺癌临床病理特征的相关性。结果浸润癌组14-3-3σ蛋白阳性率明显低于其余各组,p73α蛋白阳性率明显高于其余各组,P均〈0.05。14-3-3σ蛋白阳性表达与乳腺癌腋窝淋巴结转移、HER-2受体、组织学分级、TNM分期相关,p73α蛋白与腋窝淋巴结转移、ER受体、PR受体、TNM分期相关。14-3-3σ与p73α蛋白表达呈负相关。结论 14-3-3σ和p73α蛋白与浸润性乳腺癌的发生发展相关,两者联合检测可为乳腺癌早期诊断和治疗提供参考。     

DNA修复基因Hrad51和转移抑制基因nm23在乳腺肿瘤中的表达及生物学意义

目的 探讨DNA 修复基因Hrad51和转移抑制基因nm23 在乳腺肿瘤中的表达及生物学意义.方法 应用S-P免疫组化法,分别检测了Hrad51 蛋白和nm23蛋白在乳腺癌、乳腺纤维腺瘤、乳腺小叶增生、导管上皮不典型增生、腺癌癌旁正常组织中的表达情况.结果 (1) Hrad51蛋白和nm23蛋白在乳腺癌组的表达与乳腺纤维腺瘤、乳腺小叶增生、导管上皮不典型增生、腺癌癌旁正常组织中的表达存在显著差异(均P＜0.01);(2) 乳腺癌中hrda51蛋白的阳性表达与肿瘤组织分级呈正相关(P＜0.01),与nm23、ER、PR的阳性表达呈负相关(均P＜0.01).(3) 乳腺癌中nm23的阳性表达与肿瘤组织分级呈负相关(P＜0.01),与ER的阳性表达呈正相关(P＜0.05),与PR的阳性表达无相关性(P＞0.05).(4) 乳腺癌发生淋巴结转移与nm23、ER的阳性表达呈负相关(均P＜0.01),与Hrad51的阳性表达呈正相关(P＜0.01),与PR的阳性表达无相关性(P＞0.05).结论 (1)Hrad51蛋白在不同病变乳腺组织中呈现不同表达,在乳腺癌组织中存在高度表达;(2)nm23蛋白也在不同病变乳腺组织中呈现不同表达,在乳腺癌组织中存在低表达;(3)检测Hrad51 蛋白和nm23蛋白的表达情况有望成为评价乳腺癌患者预后的一个新指标.     

BRCA1在散发性乳腺浸润性导管癌癌组织中的表达及意义

应用免疫组化技术对10例良性乳腺病变组织及89例散发性乳腺浸润性导管癌组织进行乳腺癌易感基因(BRCA1)免疫组化染色.结果 BRCA1在良性乳腺病变组织中均呈阳性显色,94.3%(84/89)的乳腺癌组织BRCA1呈阳性反应.散发性乳腺浸润性导管癌组织中随组织学分级增高,BRCA1蛋白表达减少,且BRCA1蛋白表达与肿瘤恶性程度呈负相关.认为BRCA1 可作为判断乳腺肿瘤病变性质及程度的生物学指标.     

缺氧诱导因子1、葡萄糖转运因子1在乳腺癌中的表达及其临床意义

目的 探讨缺氧诱导因子1（HIF-1α）、葡萄糖转运因子1（Glut1）在乳腺癌中的表达及其临床意义.方法 用免疫组化法检测20例正常乳腺组织、20例不典型增生乳腺组织、60例乳腺癌组织中HIF-1α、Glutl的表达,并分析其与乳腺癌临床病理特征的关系.结果 HIF-1α、Glutl在乳腺癌和不典型增生乳腺组织中的阳性率明显高于正常乳腺组织（P＜0.05）.在乳腺癌组织中HIF-1α、Glut1 蛋白阳性表达率与组织病理学分级、临床分期及淋巴结转移有关（P＜0.05）,但与肿瘤大小无关;HIF-1α、Glut1蛋白表达在乳腺癌组织中呈正相关（P＜0.05）.结论 HIF-1α和Glutl蛋白的过表达共同参与了乳腺癌的发生、发展.     

NEK2、ERK2和P53在乳腺癌中的表达及意义

目的 通过检测乳腺原位癌及浸润性癌组织中NIMA相关蛋白激酶Nek2、细胞外信号调节激酶ERK2和细胞周期调节因子P53的表达,探讨其在乳腺癌的意义及相关性.方法 采用免疫组织化学ElivisionTMplus两步法,研究86 例乳腺浸润性癌、10例乳腺导管原位癌、20例正常乳腺组织NEK2、ERK2和P53的表达情况.结果 Nek2在三组中的表达率分别为87.2%、50.0%和10.0%;ERK2分别为96.7%、80.0%和10.0%;P53分别为0.0%、40.0%和58.1%.Nek2与ERK2的表达呈正相关,ERK2与P53的表达呈正相关,NEK2与P53的表达呈正相关.Nek2蛋白和ERK2蛋白的表达均与患者发生肿瘤的淋巴结转移、组织学分级及临床分期呈明显正相关,而与患者的年龄,肿物大小,月经情况,组织类型无关.而P53仅与组织学分级有关.结论 ERK2相关信号传导通路和Nek2、P53相关的细胞周期调节共同参与了乳腺癌的形成过程.     

胃癌组织中Bcl-2、p53蛋白表达及分析

采用免疫组化法检测Bcl-2和p53蛋白在胃癌组织中的表达.发现:Bcl-2和p53蛋白在正常胃黏膜组织不表达;Bcl-2在高分化腺癌和重度不典型增生的阳性表达率明显高于中、低度分化腺癌和中、轻度不典型增生组织,p53在重度不典型增生及低分化腺癌阳性表达率均明显高于中、轻度不典型增生和中重度分化腺癌.胃癌淋巴结转移组Bcl-2阳性率明显低于无淋巴结转移组,p53阳性率明显高于无淋巴结转移组.二者的表达呈显著负相关.认为Bcl-2和p53通过细胞凋亡参与肿瘤的发生、发展的不同价段,对判断胃癌的预后有一定的参考价值.     

环指蛋白181在乳腺浸润性导管癌中表达及其临床意义

目的研究环指蛋白(RN)181在乳腺浸润性导管癌中表达的临床病理学意义。方法采用免疫组织化学染色Elivision方法检测RN181蛋白在79例乳腺浸润性导管癌癌组织及其36例乳腺增生组织中的表达。结果 RN181表达主要位于胞质,RN81在乳腺浸润性导管癌中的表达水平显著低于乳腺增生组织(P=0. 000),且与淋巴结转移呈负相关(P=0. 000)。RN181在乳腺浸润性导管癌中的表达与年龄(P=0. 918)、肿瘤大小(P=0. 817)、组织学分级无明显相关性(P=0. 150)。RN181在4种不同乳腺癌分子分型中的表达存在显著差异性(P=0. 001),其中luminal B型中RN181的表达显著低于其他三种亚型[luminal A,人表皮生长因子受体(HER)2过表达型,三阴型](P0. 05)。结论 RN181在乳腺癌中可能作为抑癌因子发挥作用,其表达在不同分子亚型乳腺癌中存在差异性,可能成为诊断乳腺癌分子分型的指标。     

乳腺癌组织中乳腺癌1号基因启动子甲基化状态与蛋白表达的关系

目的研究乳腺癌组织中乳腺癌1号基因(BRCA1)启动子甲基化与蛋白的表达情况,并分析BRCA1启动子甲基化及蛋白表达与乳腺癌生物学行为的关系。方法应用甲基化特异性PCR(MSP)技术和免疫组织化学SP法检测48例患者乳腺癌组织、癌旁乳腺组织中BRCA1启动子甲基化及蛋白表达情况。结果乳腺癌组织中BRCA1启动子甲基化阳性率(27.1%)明显高于其癌旁乳腺组织(0,P0.05);BRCA1启动子甲基化与乳腺癌的组织学分级及淋巴结转移有关(P0.05);乳腺癌组织中BRCA1蛋白阳性率(54.2%)明显低于其癌旁乳腺组织(100%,P0.05);BRCA1蛋白的表达与乳腺癌的组织学分级及淋巴结转移有关(P0.05);BRCA1启动子甲基化与蛋白表达呈明显的负相关(P0.05)。结论BRCA1启动子的异常甲基化是引起蛋白表达缺失的原因之一,BRCA1蛋白表达缺失与乳腺癌的发生、发展有关。     

胃癌发生过程中凋亡基因Survivin与Fas表达及其与幽门螺杆菌感染的关系

目的探讨凋亡基因Survivin、Fas在胃癌发生过程中的表达、临床意义及和幽门螺杆菌（Hp）感染的关系。方法采用分子原位杂交分别检测12例正常胃黏膜、22例浅表性胃炎、25例肠上皮化生、37例异型增生及52例胃癌组织中的Survivin-mRNA和Fas-mRNA表达，并检测患者Hp感染状况。结果Survivin-mRNA在肠上皮化生、异型增生组和胃癌组织中的阳性率分别为28．0％、43．2％和69．2％。胃癌组明显高于肠化和异型增生组（P〈0．01；P〈0．05）。胃癌组Fas-mRNA阳性率为36．5％，显著低于对照组和异型增生组（P均〈0．01）。Survivin-mRNA在高分化、中分化和低分化及未分化型胃癌组织中阳性率呈现递增趋势，而且其表达和淋巴结转移、远处转移密切相关。Fas-mRNA阳性率在高、中和低分化及未分化型胃癌患者中呈现递减趋势，且其表达和淋巴结转移密切相关。异型增生患者Survivin-mRNA表达与Hp感染之间呈明显正相关（P〈0．01）。相关回归分析显示胃癌患者各病理分期中Survivin-mRNA与Fas-mRNA表达呈负相关。结论在胃黏膜癌变过程中，Survivin的表达和作用逐渐上调，而Fas的表达和作用逐渐下调，而且在癌前病变组织中Survivin表达和Hp感染有密切关系；在胃癌组织中Survivin和Fas的表达呈负相关。     

Expression of c-kit protein in proliferative lesions of human breast: sexual difference and close association with phosphotyrosine status

PURPOSE: The c-kit gene which codes transmembrane tyrosine kinase receptor protein plays an important role in several types of normal and/or neoplastic human tissues. We examined the expression patterns of c-kit protein in proliferative lesions of human breast tissues in both sexes. METHODS: The localization of c-kit protein was examined immunohistochemically in human breast, consisting of 366 normal tissue, 156 benign lesions (fibroadenoma, fibrocystic change, intraductal papilloma, benign phyllodes tumor, and gynecomastia), 13 borderline diseases (atypical ductal hyperplasia, atypical lobular hyperplasia, and borderline malignant phyllodes tumor), and 197 malignant lesions (non-invasive and/or invasive ductal carcinoma and malignant phyllodes tumor). RESULTS: In normal tissues and benign proliferative lesions, c-kit product was consistently detected on epithelial cell membranes and/or cytoplasms regardless of gender difference. In contrast, we failed to find c-kit product in female borderline epithelial lesions, including atypical lobular hyperplasia, or in female malignant lesions, except for two carcinomas. In situ hybridization analysis of c-kit mRNA in female tissues gave results comparable to those obtained by immunohistochemistry. On the other hand, c-kit product was consistently detected in male benign and malignant proliferative lesions. Apart from the female breast carcinomas which lacked c-kit, c-kit expression was almost always accompanied by positivity for phosphotyrosine in the breast tissues examined, suggesting possible phosphorylation of tyrosine residues of the c-kit receptor protein. CONCLUSIONS: Loss of c-kit product was related to malignant transformation in female breast, but not in the case of male breast. We suggest that the oncogenesis pathway of breast epithelium is different between males and females in terms of c-kit expression.     

Raf激酶抑制蛋白在乳腺癌组织中的表达及意义

目的探讨Raf激酶抑制蛋白（RKIP）在浸润性乳腺癌进展中的意义。方法应用免疫组化SP法检测RKIP在16例乳腺非典型增生性病变和48例浸润性乳腺癌原发灶及其淋巴结转移灶中的表达情况,并对RKIP表达与临床病理特征的关系行Spearman等级相关分析。结果由非典型增生性病变至乳腺癌原发灶、转移灶RKIP表达水平逐渐下降,乳腺癌淋巴结转移灶与原发灶及非典型增生性病变相比RKIP表达水平明显下降（P均〈0．05）;RKIP在乳腺癌组织中的表达与患者年龄、肿瘤大小及组织学分级均无明显相关性（P均〉0．05）。结论RKIP表达异常在浸润性乳腺癌转移过程中具有重要作用,可作为判断乳腺癌进展和预后的独立指标。     

Expression and significance of CD44s, CD44v6, and nm23 mRNA in human cancer

AIM: To investigate the relationship between the expression levels of nm23 mRNA, CD44s, and CD44v6,and oncogenesis, development and metastasis of human gastric adenocarcinoma, colorectal adenocarcinoma,intraductal carcinoma of breast, and lung cancer.METHODS: Using tissue microarray by immuhistochemical (IHC) staining and in situ hybri-dization (ISH), we examined the expression levels of nm23mRNA, CD44s, and CD44v6 in 62 specimens of human gastric adenocarcinoma and 62 specimens of colorectal adenocarcinoma; the expression of CD44s and CD44v6in 120 specimens of intraductal carcinoma of breast and 20 specimens of normal breast tissue; the expression of nm23 mRNA in 72 specimens of human lung cancer and 23 specimens of normal tissue adjacent to cancer.RESULTS: The expression of nm23 mRNA in the tissues of gastric and colorectal adenocarcinoma was not significantly different from that in the normal tissues adjacent to cancer (P＞0.05), and was not associated with the invasion of tumor and the pathology grade of adenocarcinoma (P＞0.05). However, the expression of nm23 mRNA was correlated negatively to the lymph node metastasis of gastric and colorectal adenocarcinoma (r = -0.49, P＜0.01; r = -4.93, P＜0.01). The expression of CD44s in the tissues of gastric and colorectal adenocarcinoma was significantly different from that in the normal tissues adjacent to cancer (P＜0.05;P＜0.01). CD44v6 was expressed in the tissues of gastric and colorectal adenocarcinoma only, the expression of CD44v6 was significantly associated with the lymph node metastasis, invasion and pathological grade of the tumor (r = 0.47, P＜0.01; r = 5.04, P＜0.01). CD44sand CD44v6 were expressed in intraductal carcinoma of breast, the expression of CD44s and CD44v6 was significantly associated with lymph node metastases and invasion (P＜0.01). However, neither of them was expressed in the normal breast tissue. In addition, the expression of CD44v6 was closely related to the degree of cell differentiation of intraductal carcinoma of breast (x2= 5.68, P＜0.05). The expressional level of nm23mRNA was closely related to the degree of cell differentiation (P＜0.05) and lymph node metastasis (P＜0.01), but the expression of nm23 gene was not related to sex, age, and type of histological classification (P＞0.05).CONCLUSION: Patients with overexpression of CD44s and CD44v6 and low expression of nm23 mRNA have a higher lymph node metastatic rate and invasion. In addition, overexpression of CD44v6 is closely related to the degree of cell differentiation. Detection of the three genes is able to provide a reliable index to evaluate the invasion and metastasis of tumor cells.     

Estrogen receptor β2 is inversely correlated with Ki-67 in hyperplastic and noninvasive neoplastic breast lesions

Purpose

The purpose of the study is to compare expression levels of ΕRα, ERβ1, ERβ2 and cell proliferation marker Ki-67 in normal breast and hyperplastic and noninvasive neoplastic breast lesions.

Materials and methods

Routinely processed breast tissue from 55 patients provided 65 cases of noninvasive lesions, namely, epithelial hyperplasia of usual type (HUT), apocrine metaplasia (AM), atypical hyperplasia (AH) and ductal carcinoma in situ (DCIS) and 14 cases of adjacent normal breast tissue. Expression of ERα, ERβ1 and ERβ2 were evaluated using immunohistochemistry and correlated with Ki-67 labeling index (Ki-67 LI) and menopausal status of the patients.

Results

Compared with normal breast, ERα expression increased in low to intermediate-grade DCIS (DCIS1/2) and tended to decrease in high-grade DCIS, while ERβ1 expression decreased in DCIS irrespective of grade. Mean Ki-67 LI in HUT, low to intermediate-grade DCIS and high-grade DCIS was higher than in normal breast. Higher than normal Ki-67 LI correlated with low ERβ2 expression in the whole set of cases and with high ERα expression and low ERβ2 expression in the postmenopausal cases of the subset that is generated by excluding AM and high-grade DCIS. Postmenopausal status correlated with low ERβ1 expression in the whole set and with higher than normal Ki-67 LI, high ERα expression and low ERβ1 expression in the subset.

Conclusions

These findings are in accordance with an ERα-opposing oncosuppressive role of ERβ2 in mammary carcinogenesis along the HUT-AH-DCIS1/2 pathway.     

乳腺癌组织中EGF和ALDH1A1的表达及其临床意义

目的 研究表皮生长因子(EGF)与乙醛脱氢酶(ALDH1A1)在正常乳腺组织及乳腺癌组织中的表达及意义,探讨EGF在乳腺癌干细胞演进过程中的作用.方法 采用RT-PCR和免疫组化方法分别检测24例正常乳腺组织和45例乳腺浸润性导管癌组织中EGF和ALDH1A1的表达情况.结果 RT-PCR结果显示,乳腺癌中EGF与ALDH1A1的mRNA水平高表达,在正常乳腺组织中低表达或不表达,两组比较P＜0.01.免疫组化结果显示,EGF在正常乳腺与乳腺癌中的表达分别为20.8％和75.6％,ALDH1A1在正常乳腺与乳腺癌中的表达分别为16.7％与66.4％,乳腺癌中EGF与ALDH1A1蛋白水平表达明显高于正常组织(P＜0.01).EGF与ALDH1A1两者在乳腺癌中表达存在正相关(P＜0.05).EGF的表达与发病年龄、肿瘤大小、组织学分型、有无淋巴结转移、PR表达无相关性,与ER表达呈正相关(P＜0.01);ALDH1A1的表达与发病年龄、肿瘤大小、有无淋巴结转移、ER、PR表达无相关性,与组织学分型呈正相关(P＜0.05).结论 EGF与ALDH1A1在乳腺癌组织中的表达高于正常乳腺组织,EGF与乳腺癌干细胞的发生发展关系密切.     

High stromal and epithelial human gh gene expression is associated with proliferative disorders of the mammary gland

We have demonstrated and localized human GH (hGH) gene expression in surgical specimens of normal human mammary gland and in proliferative disorders of the mammary gland of increasing severity using sensitive in situ RT-PCR methodology. hGH mRNA identical to pituitary hGH mRNA was first detected by RT-PCR of RNA derived from samples of normal human mammary gland. Cellular localization of hGH gene expression in the normal mammary gland exhibited restriction to luminal epithelial and myoepithelial cells of the ducts and to scattered stromal fibroblasts. We subsequently examined the expression of the hgh gene in three progressive proliferative disorders of the human mammary gland, i.e. A benign lesion (fibroadenoma), a pre-invasive stage (intraductal carcinoma) and an invasive ductal carcinoma. hGH mRNA was readily detected in the tumoral and non-tumoral epithelial components and also in cells of the reactive stroma including fibroblasts, myofibroblastic and myoepithelial cells, inflammatory infiltrate lymphocytes and endothelial cells in areas of neovascularization. In all three proliferative disorders examined, the intensity of the cellular labeling observed in both the epithelial and stromal compartments was always stronger compared with that in adjacent normal tissue. hGH protein was also present in significantly higher concentration in extracts derived from proliferative disorders of the mammary gland compared with extracts derived from normal mammary gland. We also examined hGH gene expression in axillary lymph nodes not containing and containing metastatic mammary carcinoma. hGH gene expression was evidenced in metastatic mammary carcinoma cells and in reactive stromal cells by both in situ hybridization and in situ RT-PCR. In contrast, in lymph nodes not containing metastatic mammary carcinoma, hGH mRNA was detected only by use of in situ RT-PCR. Thus, increased expression of the hGH gene in the epithelial component and the de novo stromal expression in proliferative disorders of the mammary gland are suggestive of a pivotal role for autocrine hGH in neoplastic progression of the mammary gland.     

热休克蛋白70 mRNA在老年人胃癌中表达的意义

目的探讨热休克蛋白70 mRNA(HSP70 mRNA)在胃癌发生中的意义.方法取胃癌44例,癌旁不典型增生20例,单纯不典型增生16例,浅表性胃炎20例.采用核酸原位杂交技术(ISH)检测HSP70 mRNA表达状况.结果在胃癌、癌旁不典型增生和单纯不典型增生中HSP70 mRNA的阳性率分别为61.36(27/44)、55.00%(11/20)和31.25%(5/16);在浅表性胃炎组阳性率为5.00%(1/20).HSP70 mRNA表达与癌细胞浸润深度和淋巴结转移无明显相关性.结论 HSP70 mRNA在胃癌形成早期有较高表达,检测HSP70 mRNA可以作为胃癌早期诊断的指标.