Immunohistochemical detection of p53 and c-erbB-2 proteins: Prognostic significance in operable breast cancer

We examined the associations of p53 expression and/or c-erbB-2 expression with Ag-NOR counts and clinicopathologic variables in 111 breast cancer patients, and assessed whether expression of either p53 or c-erbB-2 would be useful prognostic indicators. There was no significant association between p⁵³ expression and c-erbB-2 expression, but p53 expression and c-erbB-2 expression, especially in combination, were shown to be significantly associated with Ag-NOR counts and axillary lymph node metastasis. Although p⁵³ expression and c-erbB-2 expression were significant prognostic factors by univariate analysis, they did not appear to be independent prognostic factors by multivariate analysis, in which nodal status was introduced using the Cox model. When nodal status was excluded from the model, however, concurrent p⁵³ and c-erbB-2 expression did have a significant prognostic value. Therefore, it was suggested that concurrent p⁵³ and c-erbB-2 expression provides valuable prognostic information for breast cancer patients in whom axillary lymph node dissection has not been performed.     

腋窝淋巴结1～3个转移的早期乳腺癌根治术后局部复发和生存预后因素的研究

背景与目的：腋窝淋巴结1—3个阳性的早期乳腺癌进行辅助放射治疗的指征尚未明确,本研究探讨这部分患者根治术后的局部／区域复发以及生存的危险预后因素。方法：回顾性分析1998年3月至2002年3月在中山大学肿瘤防治中心接受标准或改良根治手术的217例乳腺癌病例的资料,原发肿瘤病理分期pT1期71例,pT2期146例,其中202例接受辅助化疗,51例接受辅助放疗,116例接受术后内分泌治疗。结果：中位随访时间69个月,全组的5年无局部复发生存率、无瘤生存率和总生存率分别为85．2％、81．8％和90．2％。44例出现肿瘤复发,其中21例局部／区域复发。生存分析表明,局部／区域复发患者的5年总生存率明显低于局部／区域控制的患者（61．9％vs93．6％,P〈0．0001）,患者年龄≤35岁、原发肿瘤pT2期和腋窝淋巴结转移比例≥30％是影响无局部复发生存率、无病生存率和总生存率的不良预后因素。根据这3项预后影响因素设立评分系统,发现不同分值病例的5年无局部／区域复发生存率差异具有统计学意义（P=0．0072）。在辅助化疗≥5疗程的159例患者中,接受辅助放疗的35例患者的各项生存率指标均优于未作放射治疗的患者。结论：对于腋窝淋巴结1～3个阳性的早期乳腺癌患者,年龄≤35岁、pT2期原发肿瘤和腋窝淋巴结转移比例≥30％提示术后局部／区域复发危险较高．应考虑辅助放疗。     

Prognostic significance of c-erbB-2 expression in node negative breast cancer.

The prognostic value of c-erbB-2 oncogene expression was studied retrospectively in a consecutive series of 230 node negative breast cancers, followed-up for at least 7 years after primary treatment. The expression of c-erbB-2 oncoprotein was determined on formalin-fixed paraffin-embedded tissue, using a monoclonal anti-c-erbB-2 antibody by the avidin-biotin immunoperoxidase method. Positive immunostaining was observed in 20.9% of cases, whereas strong diffuse positivity was recorded only in 8.7% of cases. C-erbB-2 gene product showed no association to T category or nuclear grade. A significant association of c-erbB-2 expression to prognosis was observed only for cases showing a strong diffuse immunostaining, but such an association was no longer statistically significant at multivariate analysis adjusting for other prognostic factors such as T category and nuclear grading. C-erbB-2 expression is of no value to predict the clinical course of node negative patients in the current practice.     

Clinical relevance of lymph node ratio in breast cancer patients with one to three positive lymph nodes

Background:

To test the hypotheses that breast cancer patients with one to three positive lymph nodes (pN1) consist of heterogeneous prognostic subsets and that the ratio of positive nodes to total nodes dissected (lymph node ratio, LNR) might discriminate patients with a higher risk as candidates for post-mastectomy radiation therapy (PMRT).

Methods:

Using information from 7741 node-positive patients, we first identified cutoff values of the LNR using the nonparametric bootstrap method. Focusing on 3477 patients with pN1 disease, we then evaluated the clinical relevance of the LNR categorised by the estimated cutoff values (categorised LNR, cLNR).

Results:

Among 3477 patients with pN1 disease, 3059 and 418 patients were assigned into the low and intermediate cLNR groups, respectively, based on a cutoff value of 0.18. The prognostic factors associated with poor overall survival (OS) included younger age, T2 stage, negative oestrogen/progesterone receptors, high histologic grade, and intermediate cLNR. Post-mastectomy radiation therapy significantly increased OS in patients assigned to the intermediate cLNR (hazard ratio, 0.39; 95% confidence interval, 0.17–0.89; P=0.0248), whereas patients in the low cLNR group derived no additional survival benefit from PMRT.

Conclusion:

This study suggests that PMRT should be recommended for patients with pN1 disease and an intermediate cLNR.     

Elevated ras oncogene expression correlates with lymph node metastases in breast cancer patients

The protein product of the ras cellular oncogene(s) (p21) was assayed in primary breast carcinomas from two groups of patients who had different axillary lymph node status. Using an immunohistochemical assay, the intensity and percent of neoplastic cells demonstrating ras p21 antigen staining were significantly higher in the primary tumors from patients with lymph nodes positive (LN+) for malignancy (20 patients) compared with the lymph node negative (LNO) group (21 patients). The expression of p21 also correlated with tumor size. Age and estrogen receptor status did not influence p21 staining. The antigen expression of p21 was similar in intensity and distribution in the primary tumor and regional lymph node metastases. Enhanced expression of p21 in primary breast cancers that metastasize to regional nodes indicates that ras p21 may be a determinant of the malignant potential of breast cancer cells and may represent a new class of more biologically relevant tumor markers.     

PTEN及bcl-2和p53蛋白表达与乳腺癌预后的关系

目的 :探讨新的抑癌基因PTEN在人乳腺癌组织中的表达、与bcl 2、p5 3的相关性及其与乳腺癌临床病理学特征和预后的关系。方法 :采用免疫组化SP法检测62例浸润性乳腺癌 (包括浸润性导管癌、浸润性小叶癌、髓样癌等 )标本中PTEN、bcl 2及p5 3蛋白的表达。结果 :62例乳腺癌标本中PTEN、bcl 2及p5 3阳性表达率分别为 64 5 % ( 4 0 /62 )、5 6 5 % ( 3 5 /62 )及2 7 4% ( 17/62 ) ;PTEN与bcl 2之间不具有相关性 ,P >0 0 5 ;PTEN与p5 3之间显著相关 ,P <0 0 1。乳腺癌PTEN蛋白表达水平与患者年龄、肿瘤组织学类型、肿瘤大小之间差异无统计学意义 ,P >0 0 5 ;与肿瘤细胞学分级、淋巴结有无转移及预后之间差异有统计学意义 ,P <0 0 5 ;PTEN低表达患者肿瘤细胞学分级高、淋巴结转移阳性率高、5年生存率低。结论 :1)抑癌基因PTEN在乳腺癌中确实存在突变或缺失。其表达水平的高低作为判断乳腺肿瘤细胞增殖活性、侵袭力及转移的重要指标。 2 )PTEN与bcl 2协同表达与预后相关 ,可作为乳腺癌预后评估的有效指标。 3 )PTEN突变或缺失可能与p5 3存在一定的相关性 ,但其作用机制有待进一步研究。     

A prospective study of the significance of gene and chromosome alterations as prognostic indicators of breast cancer patients with lymph node metastases

In 150 surgically resected primary breast carcinomas that had axillary lymph-node metastases, we examined the incidence of loss of heterozygosity on chromosomes 16p, 16q, 17p, 17q, and 18q, point mutation of the p53 tumor-suppressor gene, nuclear immunoreaction of p53 protein, and amplifications of the c-erbB-2 and int-2 oncogenes by Southern blotting, single-strand conformation polymorphism analysis, and immunohistochemistry. We analyzed the association of these factors and conventional prognostic parameters with outcome of the patients, using Cox's univariate and multivariate analyses. The univariate analysis revealed that nuclear p53 immunoreaction, p53 mutation, and c-erbB-2 amplification as well as the number of metastatic lymph nodes, histological grade, and hormone-receptor statuses were significant prognostic indicators for both recurrence and cancer death. p53 immunoreaction was correlated more strongly with a poor prognosis than p53 mutations. The combination of p53 and c-erbB-2 effectively identified the high-risk patient group, and even among Grade 3 cases the subgroup with these alterations tended to have poorer clinical outcomes. The multivariate analysis including p53, c-erbB-2, and conventional factors. Lymph node status, grade, and p53 had independent impacts on the survival of patients. Under identical adjuvant systemic therapies, prognoses differed between the patient groups with and without alterations of p53 or c-erbB-2. Appropriate combinations of conventional factors with nuclear p53 immunoreaction and c-erbB-2 amplification would help to identify highly aggressive node-positive breast carcinomas and would aid stratification of patient groups in randomized clinical trials of adjuvant systemic therapies.     

Mutant p53, EGF receptor and c-erbB-2 expression in human breast cancer.

p53 expression was studied in 111 primary breast cancers with a monoclonal antibody (PAb240) that reacts with an epitope in mutant p53. Epidermal growth factor receptors (EGFr) and oestrogen receptors (ER) measured by ligand binding, c-erbB-2 expression assessed by immunochemistry and lymph node status were compared with p53 staining. Fifty-nine tumours (53%) were positive for p53, and this correlated with EGFr expression (P less than 0.02), which is a known poor prognostic factor. Eighteen out of 59 p53+ tumours expressed c-erbB-2 versus 4 out of 52 p53- tumours assessed on paraffin sections. However, assessing c-erbB-2 by Southern blotting and immunochemistry on frozen sections showed that 20 out of 59 p53+ tumours overexpressed or had amplified c-erbB-2 compared with 15 out of 52 p53- tumours (not significant). Mean EGFr concentration in p53+ tumours was 31 fmol per mg of membrane protein versus 14 fmol mg-1 in p53- tumours. Cytoplasmic staining was the most frequent pattern found for p53, but some cases showed cytoplasmic plus nuclear staining, or focal cells with predominantly nuclear staining. Thus, abnormal p53 is the commonest oncogene abnormality described in breast cancer. The association with EGFr expression suggests that these oncogenes interact in the pathogenesis of breast cancer.     

Loss of hMSH2 expression in primary breast cancer with p53 alterations

Inactivation of DNA mismatch repair genes (MRG) is a recently described pathway of cancer development and progression resulting in genetic instability. Germline mutations in MRG have been studied predominantly in patients with hereditary non-polyposis colorectal cancer (HNPCC) where it is associated with microsatellite instability (MSI). The expression of MRG in primary breast cancer is still largely unexplored. The hMSH2 MRG encodes a protein that recognizes and binds to mismatch sequences of DNA. We investigated the relation-ship between hMSH2 expression and clinicopathological and biological characteristics, including p53 and p185 expression, in 44 primary invasive breast cancers. hMSH2 was not expressed in 11 cases (25%). Interestingly, p53 (p=0.05), p185 and steroidal receptor expression (p=0.07) were more frequent in tumors without hMSH2 expression. Furthermore, in 30 of 44 cases we analyzed hMSH2 expression in relation to MSI at 9 dinucleotide loci, and found that MRG expression was not significantly related to MSI. The presence of hMSH2 and p53 alterations in the same tumor suggests that the two oncoproteins act through a common mutational pathway, whereas the absence of a correlation between hMSH2 and MSI suggests that oncogenetic mechanisms of progression in primary breast cancer differ from those in HNPCC.     

乳腺癌肿瘤代谢异质性与p53基因表达和淋巴结转移相关性的研究

目的:分析乳腺癌肿瘤代谢异质性与p53基因表达和乳腺癌淋巴结转移的关系。方法:选取接受18F-FDG PET/CT检查的原发性乳腺癌患者34例,分别测定每例患者原发灶的肿瘤代谢异质因子（heterogeneity factor,HF）,同时将HF与p53基因表达和淋巴结转移进行相关性分析,采用受试者工作特性曲线（area under receiver operating characteristic curve,ROC）分析HF对p53基因突变及淋巴结转移的诊断效能。结果:HF与乳腺癌p53基因表达呈弱相关性（P<0.05）;HF与淋巴结转移无相关性（P>0.05)。结论:HF与乳腺癌p53基因表达具有相关性,HF对预测p53基因表达具有一定意义,然而,HF不能预测乳腺癌是否有淋巴结转移。     

Diagnosis of axillary lymph node metastases in patients with breast cancer

Summary The diagnosis of axillary (AX) metastases remains a challenge in the management of breast cancer and is a subject of controversy. Clinical node staging clearly is limited in the assessment of AX lymph nodes. AX mammography, ultrasonography, and computed tomography (CT) do not provide histologic information. Although nuclear magnetic resonance imaging may have considerable value in the diagnosis of AX metastases, it does not detect micrometastases. The use of biologic markers in the assessment of AX metastases remains a subject of investigation. On the other hand, biopsy of selected AX nodes or tissue with examination of histology or cytology generally would not identify a significant percentage of patients with AX node involvement. Sentinel lymph node biopsy, however, might be potentially useful for assessing AX metastases, although it remains investigational. In order to simplify diagnosis and reduce morbidity and mortality, alternatives to AX dissection must be sought and imaging and staging modalities refined. We present a review of the literature pertaining to the diagnosis of AX metastases in patients with breast cancer and a discussion of some current areas of controversy.     

Prognostic significance of p53, bax and bcl-2 gene expression in patients with laryngeal carcinoma.

AIM: This study was designed to examine the prognostic significance of the coexpression of three genes (bax, bcl-2 and p53) which play a critical role in the apoptotic mechanisms in patients with squamous cell laryngeal carcinoma. MATERIALS AND METHODS: The immunohistochemical expression of bcl-2, bax and p53 genes was retrospectively examined in 38 patients with squamous cell laryngeal carcinoma and in five controls (necrotomic tissue). Tissue specimens were obtained both during the diagnostic biopsy and at the time of surgery. Clinicopathological and survival data were correlated with the staining results. RESULTS: Bcl-2 protein expression (P=0.0472), stage (P=0.0087) and lymph-node involvement (P=0.0488) were found to be independent prognostic factors. Increased bcl-2 protein expression correlated with a better 5-year survival (P=0.0472). Patients who were bcl-2(-)/p53(-) (n=25) or bax(+)/bcl-2(-) (n=13) had a significantly worse overall survival (P=0.0305 and P=0.0482, respectively). Similarly, patients who were bax(+)/bcl-2(-)/p53(-) (n=11) also had a worse 5-year survival compared with the rest of the group (P=0.0088). Changes that were noticed in bax and p53 protein expression from the time of biopsy until the time of surgery did not correlate with a significant increase in the overall survival. CONCLUSIONS: The expression of bcl-2 gene appears to be an independent prognostic factor for patients with laryngeal carcinoma. The coexpression of the genes studied can be used to determine aggressive clinical phenotypes.     

颈胸联合根治术在伴有锁骨上淋巴结转移的乳腺癌患者中的应用

目的:结合新辅助化疗探讨颈、胸联合根治术在伴有锁骨上淋巴结转移的乳腺癌患者中的应用。方法:选择12例伴有锁骨上淋巴结转移的乳腺癌患者在术前行2～3周化疗(CAF或CMF方案),发现原发肿瘤及锁骨上淋巴结均缩小后,同时施行乳腺癌根治术和锁骨上淋巴结清除术,术后继续给予综合治疗。结果:联合根治术后,全部病例无手术并发症出现,无肉眼肿瘤残留。共切除锁骨上淋巴结142枚,平均11.8枚,其中转移116枚,平均9.7枚。最多的患者淋巴结切除20/21(+),平均转移率达81.7%。乳腺腋窝清除术中切除淋巴结290枚,平均24.1枚,其中转移204枚。1例患者的病理诊断为右乳腺内下及中下髓样癌,腋下淋巴结为0/19,而锁骨上淋巴结清除术后转移情况为1/4(+),属于跳跃性转移。结论:结合新辅助化疗实行联合根治术,可以最大限度地切除肿瘤及转移淋巴结。     

颈胸联合根治术在伴有锁骨上淋巴结转移的乳腺癌患者中的应用

目的：结合新辅助化疗探讨颈、胸联合根治术在伴有锁骨上淋巴结转移的乳腺癌患者中的应用.方法：选择12例伴有锁骨上淋巴结转移的乳腺癌患者在术前行2～3周化疗（CAF或CMF方案）,发现原发肿瘤及锁骨上淋巴结均缩小后,同时施行乳腺癌根治术和锁骨上淋巴结清除术,术后继续给予综合治疗.结果：联合根治术后,全部病例无手术并发症出现,无肉眼肿瘤残留.共切除锁骨上淋巴结142枚,平均11.8枚,其中转移116枚,平均9.7枚.最多的患者淋巴结切除20/21（＋）,平均转移率达81.7%.乳腺腋窝清除术中切除淋巴结290枚,平均24.1枚,其中转移204枚.1例患者的病理诊断为右乳腺内下及中下髓样癌,腋下淋巴结为0/19,而锁骨上淋巴结清除术后转移情况为1/4（＋）,属于跳跃性转移.结论：结合新辅助化疗实行联合根治术,可以最大限度地切除肿瘤及转移淋巴结.     

1至3个腋窝淋巴结转移乳腺癌根治术后放疗的应用

Nowadays the decision whether to offer early breast cancer patients with 1～3 positive axillary nodes adjuvant radiotherapy is a heated controversy. Results of several randomized trials have shown that post-mastectomy radiotherapy including the whole chest wall and regional lymph node-bearing areas can improve the Iocoregional control and survival rate of patients with 1～3 positive axillary nodes. Tumor size and stage, number of dissected nodes, positive lymph node ratio, extracapsular extension and lymphovascular invasion may determine whether post-mastectomy radiotherapy is needed or not for these patients. The best radiotherapy approach and patients selections should be verified by further prospective randomized control clinical trials.     

1至3个腋窝淋巴结转移乳腺癌根治术后放疗的应用

Nowadays the decision whether to offer early breast cancer patients with 1～3 positive axillary nodes adjuvant radiotherapy is a heated controversy. Results of several randomized trials have shown that post-mastectomy radiotherapy including the whole chest wall and regional lymph node-bearing areas can improve the Iocoregional control and survival rate of patients with 1～3 positive axillary nodes. Tumor size and stage, number of dissected nodes, positive lymph node ratio, extracapsular extension and lymphovascular invasion may determine whether post-mastectomy radiotherapy is needed or not for these patients. The best radiotherapy approach and patients selections should be verified by further prospective randomized control clinical trials.     

1至3个腋窝淋巴结转移乳腺癌根治术后放疗的应用

Nowadays the decision whether to offer early breast cancer patients with 1～3 positive axillary nodes adjuvant radiotherapy is a heated controversy. Results of several randomized trials have shown that post-mastectomy radiotherapy including the whole chest wall and regional lymph node-bearing areas can improve the Iocoregional control and survival rate of patients with 1～3 positive axillary nodes. Tumor size and stage, number of dissected nodes, positive lymph node ratio, extracapsular extension and lymphovascular invasion may determine whether post-mastectomy radiotherapy is needed or not for these patients. The best radiotherapy approach and patients selections should be verified by further prospective randomized control clinical trials.     

1至3个腋窝淋巴结转移乳腺癌根治术后放疗的应用

Nowadays the decision whether to offer early breast cancer patients with 1～3 positive axillary nodes adjuvant radiotherapy is a heated controversy. Results of several randomized trials have shown that post-mastectomy radiotherapy including the whole chest wall and regional lymph node-bearing areas can improve the Iocoregional control and survival rate of patients with 1～3 positive axillary nodes. Tumor size and stage, number of dissected nodes, positive lymph node ratio, extracapsular extension and lymphovascular invasion may determine whether post-mastectomy radiotherapy is needed or not for these patients. The best radiotherapy approach and patients selections should be verified by further prospective randomized control clinical trials.     

1至3个腋窝淋巴结转移乳腺癌根治术后放疗的应用

Nowadays the decision whether to offer early breast cancer patients with 1～3 positive axillary nodes adjuvant radiotherapy is a heated controversy. Results of several randomized trials have shown that post-mastectomy radiotherapy including the whole chest wall and regional lymph node-bearing areas can improve the Iocoregional control and survival rate of patients with 1～3 positive axillary nodes. Tumor size and stage, number of dissected nodes, positive lymph node ratio, extracapsular extension and lymphovascular invasion may determine whether post-mastectomy radiotherapy is needed or not for these patients. The best radiotherapy approach and patients selections should be verified by further prospective randomized control clinical trials.