生物学标记物应用于肝癌原位肝移植预后的研究

目的：探讨生物学标记物对预测肝癌移植术后复发的价值。方法:回顾性分析2002年4月至2005年11月50例肝癌移植病例，存活者至少随访12月，石蜡切片做免疫组化检测E-cadherin、β-catenin、Ki-67的表达，分析其与复发、生存时间的相关性并与一些主要临床病理指标进行比较。结果:TNM分期、门静脉癌栓仅在单变量分析中对预测复发、生存时间有意义（P<0.05），术前血清AFP仅在多变量分析中与复发有关（OR=2.552，P<0.05）；胞膜E-cadherin低表达、核β-catenin阳性、核Ki-67高增殖指数在单变量分析中对预测复发、生存时间的统计学意义均显著，而且在多变量分析中结果类似（均P<0.01）,但胞膜β-catenin表达和预后无关(P>0.05)。结论:这3个生物学标记物有前景成为肝癌移植术后复发的预后因子，且价值可能优于临床病理指标。     

Tumoral and angiogenesis factors in hepatocellular carcinoma after locoregional therapy

Locoregional therapy (LRT) is used as a bridge to orthotopic liver transplant (OLT) for hepatocellular carcinoma (HCC) patients. Liver explants in OLT patients with HCC were studied regarding both tumor stage, histology, and immunohistochemical staining for cytokeratin (CK)7, CK19, P53, Ki-67, and vascular endothelial growth factor (VEGF). Patients receiving no LRT (control) (n=30) were compared with LRT treatment groups with conventional transarterial chemoembolization (cTACE) (n=25) or drug-eluting bead transarterial chemoembolization (DEB TACE) (n=17). Tumor stage and histology were similar between treatment and control groups. The mean percent necrosis was significantly higher for treatment groups versus the control group (p<0.0001 for both groups versus control) and was significantly higher in the cTACE group versus the DEB TACE group. Only the DEB TACE group showed peritumoral CK19 positivity, and tumors were all CK19-negative. Using a threshold of 50% of tumoral cells, tumoral VEGF was significantly different between groups, with the control group having the highest degree of positivity; however, peritumoral VEGF was not significantly different between the groups. The Ki-67 proliferation fraction was higher in the treated groups with a statistically significant difference between the DEB-treated group and those without treatment (p=0.02). There were no statistically significant differences in tumoral or peritumoral CK7 or p53. Percent necrosis and percent Ki-67 positivity were higher with LRT, with a significant difference between groups for percent necrosis, confirming that LRT causes necrosis and suggesting that treatment leads to increased proliferation and decreased tumoral VEGF.     

Clinical characterization for proliferation and metastasis in advanced hepatocellular carcinoma patients

Here we reported that association between drug resistance and carcinomatosis in advanced liver cancer cases. All subjects (n=4) were periodically received chemotherapeutic agents when clinical manifestation being defined serologically. Hepatic specimen was harvested via biopsy and further prepared as paraffin-slice before conducting immunohistochemistry. As a consequence, more detectable biomarkers, such as AST, AFP, GGT2, were high expressed in plasma when compared to clinical standards. DNA topoisomerase II (TOPO II), Ki-67 were immunoreactively labeled in cytoplasm/membrane and nucleolus of liver cancer cells, while hepatocellular tumor protein p53 was negative or non-detected. Additionally, we found that hepatobiliary cancer showed epithelial differentiation with pronounced CK19 immunoreactivity when metastasizing. Our clinicopathologic findings demonstrate that correlation between carcinomatous proliferation/metastasis and drug resistance protein expression. Furthermore, these evidences indicate that TOPO II may be a biomarker for advanced hepatocellular carcinoma patient receiving chemotherapeutics.     

Expression of cyclooxygenase-2, p53 and Ki-67 in gastric cancer

It has been reported that p53 mutation may contribute to upregulate cyclooxygenase (COX)-2 expression that is observed in malignant tissues. These molecules are involved in carcinogenesis by affecting tumor cell proliferation. The aim of this study was to examine the relationship between COX-2 or p53 expression and clinico-pathological characteristics including tumor cell proliferation in gastric cancer. COX-2 and p53 expressions were investigated with immunostaining, in tissue specimens obtained from 119 patients who underwent surgery for gastric cancer. The Ki-67 labeling index (LI) was counted by Ki-67 immunostaining. COX-2 and p53 expressions correlated significantly with depth of tumor invasion. However, there was no association between COX-2 or p53 expression and survival. p53 expression did not correlate with COX-2 expression. There was no significant difference in various clinicopathological variables between Ki-67 LI subgroups. The mean Ki-67 LI value of COX-2 positive tumors was significantly higher than that of negative tumors. The mean Ki-67 LI value of p53 positive tumors was not significantly higher than that of negative tumors. The mean Ki-67 LI value of both COX-2 and p53 positive tumors was significantly higher than that of both negative tumors. These results imply that COX-2 expression is associated with tumor cell proliferation of gastric cancer.     

肝癌肝移植患者外周血中AFP mRNA和MXR7 mRNA的表达及其与预后的关系

目的： 探讨甲胎蛋白（ɑ-fetoprotein,AFP）mRNA和米托蒽醌抗性基因（Mitoxantrone-resistant 7，MXR7）mRNA作为游离癌细胞（isolated tumor cells,ITC）的标志物在肝癌肝移植患者外周血中的表达情况及其与术后肿瘤复发和转移的关系。方法: 以2002年4月至2003年12月期间的53例肝癌肝移植患者为对象，通过建立稳定可靠的实时荧光定量RT-PCR检测方法来定量检测肝癌患者外周血全细胞中AFP mRNA和MXR7 mRNA在整个肝移植术围手术期的表达和动态变化情况，并与术后肝癌复发和转移进行相关分析。 结果: 将研究对象分为肝癌移植组（53例）、晚期肝癌组（8例）、良性肝病组（26例）、正常对照组（10例），检测发现在肝移植术前，晚期肝癌组的AFP mRNA和MXR7 mRNA其表达率均达到100％,在肝癌移植组中表达率分别为57.7%和53.6%。2者均明显高于在良性肝病组（表达率为19.2%和0）和正常对照组（表达率均为0）（P<0.05）。同时在2者的阳性结果表达水平上，晚期肝癌组和肝癌组要明显高于良性肝病组和正常对照组(P<0.05)，前2者阳性结果定量水平达到1×10¹²copies/g RNA，是后2者定量水平的100－200倍。围手术期AFP mRNA和MXR7 mRNA的表达无论是表达率还是阳性结果表达水平，均以在术中的检测标本为最高，而术后1周为次之。术后出现2次以上的持续性的AFP mRNA和MXR7 mRNA的表达往往提示肝癌的复发和转移，较之移植术后没表达或一过性升高的病例，其差异有统计学意义（P<0.05），而术前和术中单次的AFP mRNA和MXR7 mRNA的表达与否则对术后肝癌复发和转移无明显影响（P>0.05）。结论: AFP mRNA和MXR7 mRNA可以作为肝癌ITC细胞的标志物，具有较好的特异性。术后血全细胞检测出现2次以上的持续性的AFP mRNA和MXR7 mRNA的表达可以有效预测肝癌肝移植术后肿瘤复发和转移。     

Combined hepatocellular carcinoma and osteoclast-like giant cell tumor of the liver: Possible clue to histogenesis

Hepatic giant cell tumor is extremely rare, and only five cases have been reported of overt hepatocellular carcinoma, thus its histogenesis is controversial. Herein is reported a case of simultaneous hepatocellular carcinoma and osteoclast-like giant cell tumor in a single tumor. A liver tumor was found in a 74-year-old woman. Histologically the tumor consisted of two distinct components: mononuclear and multinuclear giant cells with osteoclastic giant cells, and a conventional hepatocellular carcinoma. The boundary between the two components showed transitional features. Immunohistochemistry showed that the osteoclast-like giant cells were CD68 and vimentin positive, but cytokeratin and AFP negative, while spindle-shaped cells were positive only for vimentin. In a portion of the hepatocellular carcinoma the cells were cytokeratin-8 and AFP positive. Ki-67 positivity was 10% for the hepatocellular carcinoma, 60% for the spindle-shaped cells, and 0% for the giant cells. It is possible that the tumor might have had a hepatocellular carcinoma origin, given the more highly proliferative sarcomatous changes and reactive osteoclast-like cells. This case provides a clue to the histogenesis of hepatic giant cell tumors.     

Prognostic value of the proliferative index determined by Ki-67 immunostaining in superficial bladder tumors

The biological behavior of urothelial carcinomas remains unpredictable. The objective of this study was to determine the prognostic value of Ki-67 index in superficial papillary bladder tumors and to correlate it with the S-phase fraction (SPF) measured by flow cytometry. Three hundred nineteen patients with newly diagnosed superficial (pTa, pT1) bladder tumors were included between September 1990 and April 1992. Patients with bladder carcinoma in situ alone were excluded. We observed 255 pTa tumors and 64 pT1 tumors, whereas 111 lesions were classified as grade G1 and 208 as grade G2-G3. Ki-67 immunostaining was performed on paraffin-embedded material using a 3-step immunoperoxidase procedure with the murine monoclonal antibody MiB1. The relation between Ki-67 expression and prognostic variables (stage, grade, tumor size, multifocality, age, and sex) was investigated by the chi-square test. Cox regression was used to describe the association between Ki-67 and tumor recurrence in 308 patients with follow-up while adjusting for potentially confounding prognostic variables. The frequency of high Ki-67 expression (> or =10%) increased with stage (P = .005) and grade (P = .001), but not with tumor size or multifocality. Two hundred one patients experienced tumor recurrence in a median follow-up of 68 months. Stage, grade, tumor size, and multifocality were all independent predictors of recurrence. Ki-67 index greater than 10% was found to be an independent predictor of tumor recurrence among patients with tumors larger than 3 cm in diameter (HR = 2.05, CI = 1.18-3.55), but not those with smaller size tumors. With regards to the DNA index, a significant but weak correlation was observed between Ki-67 expression and the SPF (Spearman's correlation coefficient = 0.23, P = .004). In addition, aneuploid tumors had significantly higher expression of Ki-67 (22.5%) than diploid tumors (10.1%) (P = .0006). Moreover, patients with DNA aneuploid bladder tumors were more likely to have more than 10% Ki-67-positive cells than those with diploid tumors. In patients with newly diagnosed pTa or pT1 bladder tumors, a Ki-67 index above 10% is an independent predictor of shorter time to recurrence only in those with tumors larger than 3 cm.     

Impact of p53 and Ki-67 in predicting recurrence and progression of superficial (pTa and pT1) urothelial cell carcinomas of urinary bladder

In predicting the aggressive behavior of bladder tumors, the histopathological characteristics of grade and invasive stage are of principal importance. However, for predicting tumor recurrence and progression, these are sufficient only to a limited extent, particularly in the case of superficial (pTa and pT1) urothelial cell carcinomas. New prognostic factors are therefore needed to avoid either insufficient or excessive treatment. In this retrospective study, we investigated the prognostic value of the p53 and Ki-67 immunoreactivity indices. The present study included 118 superficial urinary bladder tumors consisting of 58 recurrent and 60 non-recurrent cases. Twenty of the recurrent tumors progressed into a higher grade and/or invasive stage. Paraffin immunohistochemical analysis was carried out using anti-p53 and anti-Ki-67 antibodies on the initial tumor tissues. We concluded that there is a highly significant relationship between the p53 and Ki-67 immunoreactivities and the histological grade and pathological stage of the tumors (P < 0.0001). We observed a significant relationship between the presence of recurrence and progression and the p53 immunoreactivity index (P < 0.01 and P = 0.017, respectively) and Ki-67 immunoreactivity index (P < 0.0001 and P = 0.046, respectively). Positivity for p53 and Ki-67 can demonstrate the risk of recurrence (p53: sensitivity = 76%, specificity = 58%; Ki-67: sensitivity = 86%, specificity = 48%) and progression (p53: sensitivity = 80%, specificity = 46%; Ki-67: sensitivity = 85%, specificity = 36%; ). We believe that both of these immunohistochemical markers can be considered valuable in addition to classical histopathological prognostic parameters for predicting recurrence and progression risks.     

Alpha-fetoprotein producing renal cell carcinoma: a case report.

We report a case of alpha-fetoprotein (AFP) producing renal cell carcinoma. A 53-year-old man with fever was found to have a left renal mass on computed tomography. No mass was detected in the liver. Serum AFP was 1,460 ng/ml. Radical nephrectomy showed a 10 cm mass in the upper half. A half of the tumor was whitish yellow and firm whereas another half was soft and bright yellow with hemorrhagic and necrotic areas. Histologically, the two areas were different. The lower part consisted of the clear cell renal cell carcinoma and the upper part consisted of granular cells. On immunohistochemistry, the granular tumor cells only were positive for AFP. Serum AFP level dropped abruptly to 383 ng/ml on the 6th postoperative day and gradually returned to normal during the 6 months. Multiple metastatic nodules were found in the lungs, liver and bone in 9th postoperative month and the AFP was less than 1 ng/ml. This suggest metastatic lesions are non-AFP producing clear cell type. It can be concluded that serum AFP elevation was due to synthesis by the renal cell carcinoma in the absence of liver neoplasm. Although AFP producing renal cell carcinoma is a rare entity, serum AFP can be a useful marker for the detection of the tumor.     

p53 but not erbB-2 expression is associated with rapid tumor proliferation in urinary bladder cancer

Tumor proliferation in bladder cancer is associated with tumor behavior. To assess the association between Ki-67 labeling index (LI), p53, and c-erbB-2 overexpression, formalin-fixed tissue samples of 160 patients with transitional cell carcinoma (TCC) of the urinary bladder were studied by immunohistochemistry. Ki-67 LI was strongly associated with tumor stage (P < .0001), tumor grade (P < .0001), and p53 status (P = .0014) but not with erbB-2 overexpression (P > .2). Ki-67 LI was higher in p53-positive tumors (19%) than in p53-negative tumors (14%) when all stages were compared. Ki-67 LI was independent of p53 expression in pTa tumors (p53-positive, 9%; p53-negative, 11%), showing that p53 overexpression alone is not sufficient to induce rapid tumor cell proliferation in pTa tumors. Ki-67 LI also was independent of p53 expression in pT2 to pT4 tumors (p53-positive, 20%; p53-negative, 23%), indicating that p53 expression is not necessary for rapid tumor cell proliferation in advanced stages. However, there was a striking difference in Ki-67 LI between p53-positive pT1 tumors (22.0% ± 8.8 standard deviation [SD]; n = 20) and p53-negative pT1 tumors (9.7 ± 8.3 SD; n = 22; P = .0001). These results suggest that increased proliferation in p53-positive pT1 tumors is caused by additional alterations that occur during tumor progression.     

Prognostic significance of the infiltrative pattern invasion in endometrioid adenocarcinoma of the endometrium

The prognostic significance of the invasive type of carcinoma cells in endometrial carcinoma is not defined. We evaluated the prognostic significance of the invasive type, as well as the immunostains of p53, c-erbB-2, Ki-67 antigen and MDM2 in endometrial endometrioid adenocarcinoma. This prospective analysis comprised 112 patients with endometrioid adenocarcinoma of the uterine corpus who had undergone surgery and were traced for more than 5 years after the operation. They were divided into recurrence (16 patients) and non-recurrence (96 patients) groups. The invasive type of carcinoma cells was divided into expansile, mixed (expansile and infiltrative) and infiltrative pattern. The difference in the invasive type (P < 0.001) and p53 expression (P = 0.004) between the recurrence and non-recurrence groups was significant in the univariate analysis. Moreover, the invasive type was significant in the multivariate analysis (P = 0.004). In contrast, the difference in MDM2 expression, c-erbB-2 expression and the Ki-67 labeling index in both groups was not significant in the univariate analysis. The infiltrative pattern of the invasive type (P < 0.001) and p53 expression (P = 0.043) were significantly related to a poor prognosis in the Kaplan-Meier method using the log-rank test. In conclusion, the current study indicated that the infiltrative pattern of the carcinoma cells is a predictor for poor prognosis in endometrioid adenocarcinoma in the uterine corpus. It was also indicated that p53 immunostains are useful as a predictor, but Ki-67 antigen, c-erbB-2 and MDM2 stains are not.     

p53 and c-erbB-2 protein expression in breast carcinomas. An immunohistochemical study including correlations with receptor status, proliferation markers, and clinical stage in human breast cancer.

Receptor status, proliferative activity, loss of differentiation, inactivation of tumor suppressor genes, and overexpression of oncogenes are related events that may affect the prognosis of patients with breast cancer. Ninety-seven unselected breast carcinomas were immunostained for estrogen and progesterone receptors, Ki-67 proliferation-associated antigen, p53 tumor suppressor gene product (p53), and c-erbB-2 protein. Immunohistochemical results and clinical data were compared. Altered p53 expression (regarded as indirect indication of inactivating gene alterations) was found in 25.8% of cases and was associated with a high Ki-67 labeling index, high mitotic count, and high histologic grade, with c-erbB-2 overexpression, and with negative estrogen and progesterone receptor status. p53 immunostaining could be found also in cytologic samples and correlated with p53 immunoreactivity on frozen sections of the corresponding tumors. c-erbB-2 protein overexpression was seen in 24.7% of cases and was associated with p53 altered expression and negative receptor status. Double immunohistochemical staining showed p53 and c-erbB-2 immunoreactivity in the same cells. Median and mean +/- standard deviation Ki-67 labeling index values were 15 and 16.32 +/- 10.05, respectively. Ki-67 labeling index was correlated with high mitotic count and was positively associated with histologic grade, negative progesterone receptor status, and p53 expression. Estrogen receptor status was not associated with any histologic or clinical parameters, whereas progesterone receptor status was associated with grading. The direct relation of p53 protein alterations with c-erbB-2 overexpression may be interpreted in light of the multistep model of tumor progression. Cases with altered expression of both p53 and c-erbB-2 proteins could be interpreted as having lost one inhibitory control mechanism of cell proliferation and having gained one activator of the malignant potential. However, in comparing cases with the p53 + c-erbB-2 + phenotype with cases showing positivity for only one of these gene products, no association with higher stages was seen. Detection of p53 altered expression on cytologic samples of malignant tumors may have diagnostic relevance, and p53 immunostaining may prove to be an additional diagnostic criterion in cytologic diagnosis.     

Expression profiling of colorectal carcinomas using tissue microarrays: cell cycle regulatory proteins p21, p27, and p53 as immunohistochemical prognostic markers in univariate and multivariate analysis.

With the rapidly growing understanding of tumor biology, molecular staging of cancer is expected to improve prognostication. This would be particularly important for cancers amenable to adjuvant treatment, such as colorectal carcinomas. To generate data for this, the tissue microarray technique may prove useful. Tissue microarrays were constructed with triplicate cores (0.6 mm diameter) from the invasive margins of a consecutive single-institution series of 184 colorectal carcinomas. Immunostaining for p53, p21, p27, Ecadherin, and beta-catenin was scored. Tumor cell proliferation was assessed by mitotic indices and Ki-67 labeling, apoptosis by quantification of apoptotic bodies. Reduced nuclear immunostaining for p21 (<10%) and p27 (< or =50%) and reduced membranous expression of Ecadherin were significantly associated with a poorer clinical course by univariate analysis. beta-catenin immunostaining had no prognostic impact. Mitotic and apoptotic indices as well as Ki-67 labeling below the median were indicators of poor prognosis. Complete absence of p53 nuclear staining was a significant adverse prognostic factor. By Cox regression, p53 = 0%, p53 = 0%, in combination with p27 < or = 50%, the mitotic index and the combined mitotic and apoptotic index added prognostic information to UICC stage. The authors found that growth pattern, lymphohistiocytic response, lymphatic permeation, and venous spread, too, each was a strong prognosticator in addition to UICC stage. The results support that tissue microarrays are a useful tool for screening immunohistochemical markers for prognostic use. An immunopanel of p21, p27, and p53 could be useful for prognostication in colorectal carcinoma in addition to UICC stage.     

KI-67 antigen expression predicts survival and correlates with histologic subtype in the WHO classification of thymic epithelial tumors

We performed an immunohistochemical study with monoclonal antibodies to Ki-67 antigen and p53 protein on 45 cases of thymic epithelial tumors classified according to the recent World Health Organization (WHO) classification system to evaluate whether there is correlation between the expression of these markers and prognosis, histologic subtype, and myasthenia gravis (MG). We also correlated histologic subtype with sex, age, MG, and survival. Ki-67 and p53 labeling indices (LIs) were expressed as a percentage of positive nuclear immunostaining by counting 1,000 epithelial tumor cells. Statistically significant differences were found between Ki-67 LI and survival (p = 0.007), whereas the prognostic implication of p53 could not be demonstrated, although there appeared a trend that patients with tumors of higher LIs had worse survival. Significant correlations were also found between Ki-67 (p < 0.0005) and p53 (p < 0.0005) LIs and histologic subtypes. No correlation was found between these parameters and MG. Histologic subtypes of the WHO classification also correlated with survival (p = 0.01), whereas no correlation was found with sex, age, and MG. In conclusion, our results indicate that the proliferative activity, assessed by Ki-67 LI, and the histologic pattern, according to WHO classification system, seems to represent reliable parameters in the prognosis of thymic epithelial tumors.     

Urine diacetylspermine as a novel tumor marker

A urine tumor marker, diacetylspermine, was examined in patients with recurrent pancreato-biliary carcinoma, liver tumor, lung carcinoma and gynecologic malignancies. The urine marker increased together with recurrence, suggesting a recurrence monitoring marker at the outpatient ward. Regarding hepatocellular carcinoma, the sensitivity of the urine marker was as high as conventional markers such as AFP and PIVKA II. Synchronous examination of serum and urine markers showed a higher sensitivity than the single serum or urine marker for hepatocellular carcinoma. The sensitivity for non-advanced hepatocellular carcinoma was 50%, while that for advanced hepatocellular carcinoma was 83%. The urine marker may be useful to detect non-advanced hepatocellular carcinoma. The sensitivity for lung cancer was 83% and that for Stage I or II was 82%. Concerning uterine cervical tumor, the value of the urine marker increased with the grade of dysplasia. The sensitivity for ovarian carcinoma was 100%, while that for benign ovarian tumor was 0%. These findings suggest that urine diacetylspermine is a useful tumor marker in hepatocellular carcinoma, lung cancer and gynecologic malignancy as well as pancreatobiliary carcinoma.     

Cell biological evaluation of liver cell carcinoma, dysplasia and adenoma by tissue micro-array analysis

The clinical and morphological definition of hepatocellular carcinoma (HCC), dysplasia and adenoma suffers from a lack of biological understanding. This is especially important in the histomorphological diagnosis of nodular liver lesions in needle biopsies. Therefore, we constructed a liver tissue micro-array (TMA) and evaluated 48 HCCs, 46 dysplasias, 8 adenomas, 20 cirrhotic specimens and 28 normal liver samples derived from 68 patients. Protein (over)expression by tumor suppressor genes p16, p53 and Rb1 was assessed by immunohistochemistry, the proliferative capacity was examined by immunostaining of Ki67. Further, DNA ploidy status (hyperdiploidy) was measured by fluorescent in situ hybridization (FISH) with a chromosome 1-specific repetitive DNA probe. An abnormal chromosome 1 number, i.e. the percentage of hyperdiploid cells, was 11.0, 13.7, 16.1, 23.7 and 31.3 for normal liver samples, adenomas, cirrhosis, dysplasias and HCCs, respectively. A significant difference was found for HCC versus cirrhosis (P = 0.024) or adenoma (P = 0.033), a trend (borderline significance) was seen for dysplasia versus cirrhosis (P = 0.094). Immunohistochemical protein localisation of p53 and Rb1, as well as Ki67 indicating proliferation, was clearly higher in HCC than in cirrhosis or dysplasia (all P < 0.001). Proliferation was also higher in HCC than in adenoma (P = 0.025), whereas a trend (borderline significance) was observed for Rb1 overexpression (P = 0.063). These data suggest that in the liver cell dysplasia-carcinoma pathway, changes in ploidy are followed by increased proliferation and cell biological perturbations involving p53 and Rb1. Adenomas can be distinguished from carcinomas, but not from dysplasias, based on ploidy and proliferation characteristics.     

乳腺交界性病变不典型囊性导管的临床病理学特征

目的 探讨乳腺的一种新的交界性病变,即不典型囊性导管(ACD)的临床病理学特征及其意义。方法 对200例术前没有做活检取材(仅经细胞学或细针穿刺活检诊断)的乳腺癌乳房切除材料作全乳腺切片检查,观察ACD的临床病理学特征和免疫组织化学特性。结果 ACD的发生率为22％(44／200),多发于绝经前女性(P=0.001),存在部位以乳腺癌巢周边多见;连续切片(间隔50张切片,150μm距离)可观察到ACD向非浸润性导管癌的移行;36％(16／44)伴有ACD的乳腺癌病例,癌组织p53染色阳性,其中12／16伴有的ACD也同时呈现阳性改变(P=0．001);α-平滑肌肌动蛋白染色显示ACD的肌上皮与非浸润性导管癌的肌上皮相似呈明显的受压变薄;伴有ACD的乳腺癌和不伴有ACD的乳腺癌两组在雌激素受体(ER),雄激素受体(PR),p53、c-erbB-2和Ki-67蛋白,肿瘤大小以及淋巴结转移方面没有明显的区别;44例ACD c-erbB-2染色呈阴性,Ki67标记指数也明显低于其所伴有的乳腺癌。结论 鉴于ACD与癌巢间的连续性及与癌组织p53蛋白的共同表达特性,将ACD作为乳腺的一种交界性病变具有一定的意义。     

肝细胞癌术后血清AFP、CA19-9、HGF水平对预后状况的影响

目的分析肝细胞癌术后血清甲胎蛋白(AFP)、糖类抗原19-9(CA19-9)、肝细胞生长因子(HGF)水平对预后状况的影响。方法将我院收入的82例肝癌患者作为研究对象,均进行根治术治疗并获得术后随访,收集其临床资料,记录其术后血清AFP、CA19-9、HGF水平,并分析其与患者预后的关系。结果复发组肿瘤直径、术后AFP、CA19-9和HGF水平均与未复发组存在差异(P<0.05);校正混杂因子后,复发组患者术后AFP、CA19-9、HGF水平与未复发组患者差异具有统计学意义(P<0.05);ROC曲线分析显示,当AFP≥167.95ng/mL、CA19-9≥42.63U/mL、HGF≥16.81μg/L时,其对应约登指数最大,预测肝细胞癌患者术后复发的灵敏度分别为86.36%、72.73%、77.27%,特异性分别为63.16%、68.42%、52.63%。术后3年无瘤生存率:术后AFP高水平组低于术后水平组(P<0.05),术后CA19-9高水平组低于术后低水平组(P<0.05),术后HGF高水平组低于术后低水平组(P<0.05)。结论肝细胞癌术后血清AFP、CA19-9、HGF水平均可预测患者术后复发情况,术后血清AFP、CA19-9、HGF水平较高的患者,预后多不理想。     

Detection of Ki-67 in liver biopsy specimens using monoclonal antibody to Mib-1

Ki-67 antigen was visualized in formalin-fixed, paraffin-embedded liver biopsy specimens using monoclonal antibody to Mib-1 to identify the proliferating hepatocytes. Thirty liver specimens obtained from 10 patients with chronic hepatitis (CH) or liver cirrhosis (LC) and 10 patients with hepatocellular carcinoma were studied. Liver specimens were treated with a pepsin solution or heated with autoclave or treated with microwave as a part of antigen retrieval system; then stained with an immunoperoxidase method using a monoclonal antibody to Ki-67 (Mib-1). Stable stainings were obtained in the sections treated with autoclave. Ki-67 was detected in the nuclei of hepatocytes, bile duct epithelium, fibroblast and infiltrating mononuclear cells. In patients with CH and LC, the numbers of hepatocytes positive for Ki-67 has a good co-relation with serum GPT level (p < 0.01), while has no relationship with the degree of fibrosis. The number of hepatocytes positive for Ki-67 has a good co-relation with the degree of the differentiation of hepatocellular carcinoma. Detection of proliferating hepatocytes using Mib-1 is useful to understand the degree of proliferation.     

Possible prognostic significance of p53, cyclin D1 and Ki-67 in the second primary malignancy of patients with double primary malignancies

Patients with two types of primary cancers are rare. In this study, we investigated the expression of p53, cyclin D1, and Ki-67 in the second primary malignancy. Tissue samples were obtained from the second primary cancer site of 43 patients who met the diagnostic criteria for double primary cancer. p53, cyclin D1 and Ki-67 were determined using immunohistochemistry. Categorical variables were compared using the Chi-squared test; correlation between data scores and histology was calculated using the Spearman’s rank-order correlation. The expression rates of p53, cyclin D1 and Ki-67 in the second primary malignancy site were 60.5%, 30.2% and 65.1% respectively. p53 expression showed statistically significant association with tumor occurrence interval, pathological grading and nodal metastasis (p < 0.05). Positive correlation was detected between the expression of cyclin D1 and Ki-67 and the expression of p53 (r = 0.313, p = 0.041; r = 0.319, p = 0.037, respectively). High-expressing p53 or cyclin D second primary malignancies were associated with decreased overall survival (p = 0.040 and p = 0.043, respectively). Ki-67 expression levels did not exhibit statistically significant differences in survival. In conclusion, elevated protein expression of p53, cyclin D1 and Ki-67 in the second primary malignancy is an indicator of more aggressive malignant behavior of the secondary tumor. These markers may have prognostic value in the clinical setting.