鼻咽癌中IGF-1R、p-ERK、c-fos和Ki-67蛋白的表达及意义

目的研究胰岛素样生长因子1受体（IGF-1R）蛋白与细胞增殖相关信号分子p-ERK、c-fos、Ki-67蛋白在鼻咽癌及鼻咽黏膜上皮细胞中的表达及意义。方法 SP法检测30例鼻咽癌、25例鼻咽黏膜上皮细胞中IGF-1R、p-ERK、c-fos、Ki-67蛋白的表达。结果 IGF-1R、p-ERK、c-fos和Ki-67蛋白在鼻咽癌细胞中阳性表达率分别为93.33%（28/30）、93.33%（28/30）、83.33%（25/30）、86.67%（26/30）,在鼻咽黏膜上皮细胞中阳性表达率分别为36.00%（9/25）、28.00%（7/25）、20.00%（5/25）、4.00%（1/25）,2组间差异有显著统计学意义（P〈0.01）;鼻咽癌中IGF-1R与p-ERK、c-fos、Ki-67蛋白阳性表达呈正相关（P〈0.05）;p-ERK与c-fos、Ki-67蛋白阳性表达呈正相关（P〈0.01）;c-fos和Ki-67蛋白阳性表达呈正相关（P〈0.01）。结论鼻咽癌中IGF-1R蛋白过表达参与调控细胞增殖。     

Proliferative Activity of Microvascular Cells in Glioblastomas does not Correlate with Time to Recurrence

The time to recurrence operation (TR) is a good growth parameter, in particular for glioblastomas. Recently, we have shown that Ki-67 labeling index (LI) of tumor cells has a high inverse correlation with this time interval. In the current study, the LI of microvascular cells (MVC) was examined in the same glioblastoma cases. The LI of MVC of primary and recurrent tumors had no relationship and did not show any correlation to TR. The growth fraction of MVC was significantly lower than that of tumor cells. The MVC in glioblastomas seems to have chaotical proliferation properties without any link to the tumor growth potential. This observation may have implications for anti-angiogenic therapy.     

胰岛素样生长因子受体通路对卵巢癌移植瘤中休眠细胞启动增殖的调控

[目的]观察胰岛素样生长因子（IGF）受体通路对卵巢癌休眠肿瘤细胞启动增殖的调控机制。[方法]采用免疫细胞化学法检测SKOV3-PKH26hi细胞IGF-1、IGF-2和IGF-1R的表达,酶联免疫吸附测定法（ELISA）测定细胞培养液中IGF-1和IGF-2的浓度,细胞计数和MTT法测定加入外源性IGF-1对SKOV3-PKH26hi细胞增殖的影响,流式细胞术检测细胞IGF-1R和Ki-67的表达、细胞周期和凋亡率,Western blot方法检测细胞p-AKT、p-GSK3β及cyclin B1的表达。[结果]SKOV3-PKH26hi细胞均可以表达IGF-1、IGF-2和IGF-1R;培养液中IGF-1和IGF-2的浓度随培养时间的延长显著升高（P〈0.05）;IGF-1对SKOV3-PKH26hi细胞能发挥促增殖作用;SKOV3-PKH26hi细胞IGF-1R、Ki-67、p-AKT、p-GSK3β、cyclin B1的表达量随着培养时间的延长而显著增高（P〈0.05）;随着培养时间的增加SKOV3-PKH26hi细胞G0/G1和G2/M期细胞显著减少（P〈0.05）,S期细胞显著增加（P〈0.05）,不同时间点的凋亡率差异无统计学意义（P〉0.05）。[结论]SKOV3-PKH26hi细胞存在IGF自分泌,IGF受体通路能够调控卵巢癌休眠细胞的增殖。     

Expression of survivin correlates with apoptosis, proliferation, and angiogenesis during human colorectal tumorigenesis

BACKGROUND: To identify the role of survivin, a novel inhibitor of apoptosis (IAP) in colorectal tumorigenesis, the authors investigated tissue expression of survivin in human colorectal tumors including 43 hyperplastic polyps, 171 adenomas with low dysplasia, 42 adenomas with high dysplasia, and 60 carcinomas in adenoma, and examined whether the expression of survivin correlated with tumor cell apoptosis, proliferation, and angiogenesis, which is known to initiate the imbalance between cell proliferation and apoptosis. METHODS: Immunohistochemical staining for the paraffin sections by using the monoclonal antibodies, survivin, p53, bcl-2, Ki-67, and CD34, was performed by the standard avidin-biotin-peroxidase technique. The apoptotic cells were detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling method, using an Apop Tag in situ detection kit. RESULTS: The immunoreactivity of survivin significantly increased in the transition from adenoma with low dysplasia to high dysplasia/carcinoma (P < 0.001). Similar changes in protein expression were observed for p53 but not for bcl-2, which was expressed throughout the colorectal tumorigenesis. This transition was associated with a significant decrease in the apoptotic index (AI) and significant increases in the Ki-67 labeling index (LI) and microvessel density (MVD; P < 0.001 for both). The expression of survivin inversely correlated with AI and was positively correlated with Ki-67 LI and MVD (P < 0.001 for both). CONCLUSIONS: These results suggest that, like p53, survivin plays an important role in transition from adenoma with low dysplasia to high dysplasia during human colorectal tumorigenesis.     

胸腺上皮性肿瘤中TTF1、Ki-67的表达与WHO分型的关系及临床意义

目的 观察TTF1及Ki-67在胸腺上皮性肿瘤(Thymic epithelial tumors,TET)中的表达情况,探讨其与WHO分型的关系及二者的相关性。方法 收集59例TET标本,其中诊断一致的51例,不一致8例。应用免疫组化技术检测TTF1及Ki-67在诊断一致的 51例TET及30例胸腺增生组织中的表达情况,并分析其与临床病理参数之间的关系。结果 在诊断不一致的8例TET中,4例与AB型有关。在诊断一致的51例TET中,TTF1及Ki-67在TET中的表达均显著高于胸腺增生组(P＜0.05)。TTF1仅在AB型胸腺瘤中表达(11/15),其表达与WHO分型有关。Ki-67标记指数从A型到胸腺癌逐渐增加,且在A型与胸腺癌的表达范围不重叠,其表达与WHO分型及MasaoKa分期均有关。同时TTF1及Ki-67在TET中的表达没有相关性。结论 TTF1是AB型胸腺瘤相对特异性的标记;Ki-67不仅对A型与胸腺癌中的区分有帮助,同时具有很好的指示预后的价值。     

Characterizing invading glioma cells based on IDH1-R132H and Ki-67 immunofluorescence

Glioma, the most common primary brain tumor, is characterized by proliferative-invasive growth. However, the detailed biological characteristics of invading glioma cells remain to be elucidated. A monoclonal antibody (clone HMab-1) that specifically and sensitively recognizes the isocitrate dehydrogenase-1 (IDH1) protein carrying the R132H mutation can identify invading glioma cells by immunostaining. To investigate the degree of invasion in gliomas of distinct grades and the proliferative capacity of the invading cells, immunofluorescent staining was conducted using antibodies against IDH1-R132H and Ki-67 on 11 surgical and autopsy specimens of the tumor core and the invading area. Higher numbers of IDH1-R132H-positive cells in the invading area correlated with a higher tumor grade. Double staining for IDH1-R132H and Ki-67 demonstrated that most invading cells that expressed IDH1-R132H were not stained by the Ki-67 antibody, and the ratio of Ki-67-positive cells among IDH1-R132H-positive cells was significantly lower in the invasion area than in the tumor core in all grades of glioma. These data suggest that higher grade gliomas have a greater invasive potential and that invading cells possess low proliferative capacity.     

Comprehensive immunohistochemical analyses on expression levels of hedgehog signaling molecules in breast cancers

Background

The hedgehog (Hh) signaling pathway plays important roles in cell proliferation, malignant progression, invasion and metastasis, and the expansion of cancer stem cells (CSCs). Comprehensive immunohistochemical (IHC) analyses have not yet been conducted on the expression levels of Hh signaling molecules in breast cancer tissues.

Methods

A total of 204 patients with invasive breast cancer treated in our institute were study subjects. IHC analyses on the expression levels of the Hh signaling molecules, sonic Hh (SHH), PTCH1, GLI1, GLI2, and GLI3 and the CSC-related factor, SOX2, were investigated.

Results

Positive correlations were observed among all of the Hh signaling molecules tested. SOX2 expression correlated with the expression levels of all Hh signaling molecules. SHH expression positively correlated with tumor size, the Ki-67 labeling index, histological grade, estrogen receptor negativity, progesterone receptor negativity, and HER2 positivity. GLI1 expression positively correlated with the histological grade. GLI2 expression positively correlated with the histological grade, Ki-67 labeling index, and HER2 positivity. Univariate analyses revealed that a younger age, larger tumor size, positive lymph node metastasis, higher histological grade, positive lymphatic invasion, and higher Ki-67 labeling index were related to poor relapse-free survival (RFS). The positivity of all Hh signaling molecules and SOX2 did not correlate with poor RFS. A multivariate analysis revealed that positive lymphatic invasion and a younger age were independent worse prognostic factors for RFS.

Conclusions

This comprehensive analysis demonstrated for the first time that SHH, GLI1, and GLI2 expression levels positively correlated with the malignant phenotypes of tumor cells.

     

Relationship between labeling indices of Ki-67 and BrdUrd in human malignant tumors

The relationship between labeling indices of Ki-67 reactive antigen expressed by cycling cells and BrdUrd incorporated into S-phase cells was investigated in 20 patients with malignant tumors. Both of the labeling indices varied greatly from patient to patient; the labeling index of Ki-67 ranged from 37.5% to 1.9% with an average value of 16%, and the BrdUrd labeling index ranged from 23.4% to 1.6% with an average of 9.3%. The Ki-67 labeling index was higher than the BrdUrd labeling index. In general, the values of the Ki-67 labeling index were parallel to those of the BrdUrd labeling index, and the relation Y = 1.59X + 1.15 (r = 0.89) was obtained. In human solid tumors, therefore, the growth fraction can be easily estimated from the S-phase fraction size, and vice versa.     

Determination of proliferative activities in human brain tumor specimens: a comparison of three methods

Summary Proliferative activities were determined in 72 human brain tumors, including 20 metastatic carcinomas, 41 gliomas, 8 meningiomas and 3 hematological tumors. Immunocytochemical techniques included labeling with monoclonal antibody (mAb) to bromodeoxyuridine (BrdU) which identifies S phase cells after previous in vitro BrdU incubation and paraffin embedding of fixed tissue specimens, and with mAb Ki-67 which reacts, in frozen sections, with a nuclear antigen expressed by all proliferating cells. BrdU labeling index (LI), Ki-67 LI and mitotic index (MI) correlated well with histological malignancy. Among the three proliferation indices, Ki-67 LI and BrdU LI were highly significantly correlated. With the exception of hematological malignancies, hyperbaric oxygenation of in vitro BrdU labeling did not significantly increase BrdU LI or depth of BrdU penetration into tissue. This study indicates that in vitro BrdU labeling is a useful alternative to Ki-67 immunolabeling of human brain tumor specimens. By such determination of tumor proliferation, it might be possible to design a more adequate postoperative therapy tailored to patients individually.     

Expression of Polo-Like Kinase (PLK1) in non-Hodgkin's lymphomas

Polo-like kinases (PLKs) are protein serine/threonine kinases that play important roles in cell division. Expression of PLK1 might, moreover, play a role in the pathogenesis of human neoplasms. The expression of PLK1 mRNA is closely correlated with survival in patients with malignant tumors. We investigated the expression of PLK1 in non-Hodgkin's lymphomas (NHLs) and analyzed the relationships between expression of PLK1, histological grade, and prognosis. We analyzed various types of NHLs from 118 patients using monoclonal antibodies against PLK1 and Ki-67. The levels of expression of PLK1 and Ki-67 were significantly lower in low-grade NHLs than in high-grade and intermediate-grade NHLs (P < 0.001). Moreover, when patients were grouped in terms of 5-year overall survival ( > 70%, group A; 50 - 70%, group B; 30 - 49%, group C; and < 30%, group D), levels of expression of PLK1 and Ki-67 were found to be significantly higher in group D than in group A and they were also significantly higher in group C than in group A (P < 0.001). Conversely, the level of expression, of Ki-67 was significantly lower in group D than in group C (P < 0.05). The labeling indices specific for PLK1 were generally higher than those specific for Ki-67. Once we divided all patients into two groups in terms of the expression levels, high-level expression group of PLK1 (PLK1 index of 70%) and Ki-67 (Ki-67 indices of 60%) and low-level expression, one of these markers (PLK1 index of >/= 70%, Ki-67 indices of >/= 60%) had a similar prognosis, an observation that can be explained by the fact that rapidly proliferating group is more drug-sensitive than the other. Our study demonstrates that expression of PLK1 might reflect the malignant potential of NHLs and that PLK1 might be more useful than Ki-67 for the detection of proliferative cells.     

MGMT和Ki-67在胶质母细胞瘤中的表达对ACNU化疗预后的影响

背景与目的:胶质母细胞瘤是预后极差的常见颅内恶性肿瘤,手术切除、放疗和化疗联合应用是常规治疗方法;Ki-67是肿瘤细胞生长活跃程度的标志,与胶质瘤的分级显著相关;O6-甲基鸟嘌呤DNA甲基转移酶(MGMT)是一种DNA修复蛋白,其表达影响肿瘤对化疗药的敏感性。本研究通过免疫组织化学方法对胶质母细胞瘤的Ki-67和MGMT进行检测,探讨其对胶质母细胞瘤化疗预后的影响。方法:总结39例脑胶质母细胞瘤患者的性别、年龄、术前Karnofsky评分、生存时间等;将患者手术切除标本石蜡切片进行Ki-67和MGMT的免疫组织化学染色,计算细胞核染色阳性率;多元逐步回归分析法判断Ki-67和MGMT的表达与患者生存时间的关系。结果:本组病例男22例,女17例;年龄21～75岁,平均54.0岁;生存时间6～38个月,平均19.3个月,中位生存期17.0个月。Ki-67在所有标本有不同程度的表达,表现为胞核明显染色,Ki-67阳性率4.0%～26.6%,平均10.5%。MGMT除2例外均有不同程度表达,胞浆染色较淡,胞核可见浓染,MGMT胞核阳性率0%～51.4%,平均21.2%。Ki-67阳性率与生存时间无相关性。MGMT胞核阳性率与生存时间呈负相关(P=0.002)。结论:Ki-67在胶质母细胞瘤表达与肿瘤的预后无关。MGMT在胶质母细胞瘤表达与肿瘤化疗后的预后有关,MGMT的检测对胶质母细胞瘤术后化疗可能有指导意义。     

Cyclin B1 is useful to predict occult cervical lymph node metastases in tongue carcinoma

Cyclin B1 plays an important role in regulating the G2-M transition of the cell cycle. In this study, we evaluated the immunohistochemical expression of cyclin B1 in a series of 40 oral tongue squamous cell carcinomas and analyzed the findings in relation to both clinicopathological variables and the Ki-67 labeling index. In normal squamous epithelium, cyclin B1 was localized in the nucleus and was expressed only in several cells of the basal and parabasal layers. By contrast, in dysplastic epithelium, cyclin B1 was localized in the cytoplasm and nuclei, and the numbers of cyclin B1 positive cells increased compared with normal epithelium. In tumor tissues, cyclin B1 expression was observed mainly in the cytoplasm, and the cyclin B1 labeling index ranged from 2 to 21%, with a mean of 11%. Cyclin B1 overexpression was positively correlated with occult cervical lymph node metastases and the number of mitotic cells. In addition, there was a positive relationship between labeling indices of cyclin B1 and Ki-67. These findings indicate that cyclin B1 could be a useful prognostic marker of occult cervical lymph node metastases in tongue carcinoma.     

Ki-67 Labeling is Correlated with the Time to Recurrence in Primary Glioblastomas

The Ki-67 labeling index (LI) was shown in many cut-off studies to be significantly correlated to the postoperative survival probability in gliomas in univariate and multivariate analyses. However, a direct relationship of the Ki-67 LI and the growth fraction, respectively, to clinical growth parameters was not demonstrated in a single tumor type of this group until now.We compared the Ki-67 LI of 20 primary glioblastomas and their recurrent tumors with the time to reoperation for recurrence. Regression analyses showed (1) a high reproducibility of the LIs at the second versus the first operation corresponding to an inherent growth potential of a given individual tumor and (2) a strong inverse correlation of LI to time to recurrence (TR) (r = –0.92). Additionally to a lower LI (corresponding to a smaller growth fraction), a longer cycle time could be derived in slowly growing as compared to rapidly growing tumors.A direct relationship of the Ki-67 LI to a clinical time parameter (the TR) was obtained, and a predictive significance of individual LIs in glioblastomas could be defined.     

Insulin-like growth factor-1 (IGF-1) induces WISP-2/CCN5 via multiple molecular cross-talks and is essential for mitogenic switch by IGF-1 axis in estrogen receptor-positive breast tumor cells

Previously, we have shown that the expression of Wnt-1-induced signaling protein-2 (WISP-2), also known as CCN5, can be regulated by multiple stimulants in estrogen receptor (ER)-positive breast tumor cells to exert their mitogenic action in these cells. Here, we show that insulin-like growth factor-1 (IGF-1), a strong mitogen, enhanced the expression of the WISP-2/CCN5 gene parallel with the induction of proliferation of ER-positive breast tumor cells. An additive effect was also seen in combination with estrogen. Perturbation of IGF-1-induced WISP-2/CCN5 expression by WISP-2-specific RNA interference impaired the mitogenic action of IGF-1 on ER-positive breast tumor cells. Furthermore, the studies have shown that the multiple molecular cross-talks and side-talks among IGF-1R, ER-alpha, and phosphatidylinositol 3-kinase (PI3K)/Akt signaling molecules are required to induce WISP-2/CCN5 mRNA by IGF-1 in ER-positive, noninvasive breast tumor cells. Because a pure anti-ER ICI 182,780 is not only able to suppress the up-regulation of WISP-2/CCN5 mRNA expression by IGF-1, it also suppresses the PI3K/Akt activity induced by IGF-1 in MCF-7 cells; we anticipate that the membrane ER receptor may participate in this event. Collectively, these studies propose for the first time that WISP-2/CCN5 is an integral signaling molecule in mitogenic action of IGF-1 axis in ER-positive human breast tumor cells.     

Increased expression of RelA/nuclear factor-kappa B protein correlates with colorectal tumorigenesis

OBJECTIVE: To identify the role of RelA/nuclear factor-kappa B, an important inhibitor of apoptosis in colorectal tumorigenesis, we examined the expression of RelA in normal colorectal mucosa (n = 10), colorectal adenomas (n = 30) and colorectal adenocarcinomas (n = 30). Furthermore, the association of RelA expression with tumor cell apoptosis, proliferation, and expression of Bcl-2/Bcl-x(L )was also studied. METHODS: Paraffin sections were stained with monoclonal antibodies directed against RelA, Bcl-2, Bcl-x(L), and Ki-67 to assess protein expression patterns in normal, adenomatous and colon cancer tissue. Apoptotic cells were detected by terminal deoxynucleotidyl-transferase-mediated dUTP-biotin nick end labeling (TUNEL) using an in situ detection kit. RESULTS: The results of immunohistochemical staining revealed that expression of RelA, Bcl-2, Bcl-x(L), and Ki-67 labeling index (LI) significantly increased in the transition from adenoma with low dysplasia to adenocarcinoma. This transition was associated with a significant decrease in the apoptotic index (AI) and a significant increase in the Ki-67 LI. The expression of RelA correlated inversely with the AI and correlated positively with the expression of Bcl-2, Bcl-x(L), and Ki-67 LI in the transition from low-grade dysplasia to adenocarcinoma. CONCLUSION: Our results suggest that increased expression of RelA/nuclear factor-kappa B plays an important role in the transition from colorectal adenoma with low-grade dysplasia to adenocarcinoma in the pathogenesis of colon cancer in humans.     

Expression and Clinical Significance of mTOR in Surgically Resected Non-small Cell Lung Cancer Tissues: a Case Control Study

Aims: Mammalian target of rapamycin (mTOR) is master regulator of the PI3K/Akt/mTOR pathway andplays an important role in NSCLCs. Here we characterized mRNA and protein expression levels of mTORand its functional associated molecules including PTEN, IGF-1R and 4EBP1 in surgically resected NSCLCs.Methods: Fifty-four patients with NSCLCs who underwent pulmonary resection were included in current study.mRNA levels of mTOR, PTEN, IGF-1R, and 4EBP1 were evaluated by RT-PCR and protein expression ofmTOR, PTEN, and IGF-1R by immunohistochemistry (IHC). Association of expression of the relevant moleculeswith clinical characteristics, as well as correlations between mTOR and PTEN, 4EBP1 and IGF-1R were alsoassessed. Results: The results of RT-PCR showed that in NSCLCs, the expression level of mTOR increased, whilePTEN, 4EBP1 and IGF-1R decreased. Statistical analysis indicated high IGF-1R expression was correlated withadvanced clinical stage (stage III) and PTEN expression was reversely associated with tumor size (P=0.16). Theresults of IHC showed mTOR positive staining in 51.8% of cases, while IGF-1R positive staining was found in83.3% and loss of PTEN in 46.3%. Protein expression of mTOR was correlated with its regulators, PTEN andIGF-1R, to some extent. Conclusions: Abnormal activation of mTOR signaling, high expression of IGF-1R, andloss of PTEN were observed in resected NSCLC specimens. The poor expression agreement of mTOR with itsregulators, PTEN, and IGF-1R, implied that combination strategy of mTOR inhibitors with other targets holdsignificant potential for NSCLC treatment.