Influence of norcantharidin on proliferation,proliferation-related gene proteins prolifera-ting cell nuclear antigen and Ki-67 of human gallbladder carcinoma GBC-SD cells

BACKGROUND: Gallbladder carcinoma is a highly lethal and aggressive disease with early metastasis, strong invasion and poor prognosis. Most patients with this disease are at the advanced and un-resectable stage and should be consi- dered for palliative treatment such as chemotherapy and ra- diotherapy. Unfortunately, reports of chemotherapy and radiotherapy for gallbladder carcinoma are disappointing. We investigated the influence of norcantharidin (NCTD) on proliferation, proliferation-related gene proteins PCNA and Ki-67 of human gallbladder carcinoma GBC-SD cells in vitro. METHODS: GBC-SD cell lines of human gallbladder carci- noma were cultured by the cell culture technique. The ex- periment was divided into NCTD group and control group. The tetrazolium-based colorimetric assay was used to evaluate cell growth. The streptavidin-biotin complex method was used to determine the expressions of prolifera- tion-related gene proteins PCNA and Ki-67 of human gall- bladder carcinoma GBC-SD cells. RESULTS: NCTD inhibited the growth and proliferation of GBC-SD cells from 10 mg/L or after 6 hours in a dose- and time-dependent manner, with the IC50 value of 56.18 μg/ ml at 48 hours. After treatment with NCTD, the expression of PCNA (0.932 ±0.031 vs. 0.318 ±0.023, P<0.001) and Ki-67 (0.964 ±0.092 vs. 0.297 ±0.018, P<0.001) proteins were decreased significantly. CONCLUSION: NCTD inhibits the proliferation of human gallbladder carcinoma GBC-SD cells in vitro and the expres- sion of their proliferation-related gene proteins PCNA and Ki-67.

The potential of carcinoembryonic antigen,p53,Ki-67 and glutathion Stransferase-π as clinico-histopathological markers for colorectal cancer

Objective： Colorectal cancer is one of the major contributors to cancer death worldwide. Lack of reliable colorectal cancer markers has hampered the management of these cancer patients. Our main purpose was to study the correlation between histopathological variables of colorectal adenocarcinomas and identify histopathological markers that are of prognostic value in patients with colorectal cancer. Methods： In the present study, we examined the expression of carcinoembryonic antigen （CEA）, p53, Ki-67 and glutathion Stransferase （GST） -n by using immunohistochemical staining methods in 126 colorectal carcinoma patients and evaluated the lymph node metastasis status in these patients by histopathological examination. Results： The positive rates of CEA, p53, Ki-67 and GST-π expression in the colorectal cancer tissue specimens examined were 95.23%, 55.56%, 53.38% and 82.30%, respectively. Expression of p53 and Ki-67 was significantly correlated with the Dukes stages of the tumor, with higher levels of these proteins in Dukes＇ C and D tumors than those in Dukes＇ A and B tumors. Furthermore, the expression of p53, GST-π and Ki-67 correlated with prognosis of patients with colorectal cancer. Additionally, the expression of p53 in colorectal cancer was closely related to the expression of Ki-67 and the expression of GST-π was directly correlated with that of p53. Conclusion： The expression of CEA, p53, Ki-67 and GST-π was correlated with various clinical features of patients with colorectal cancer. The combined use of these histopathological markers appeared to be a promising tool in predicting the prognosis of patients with this type of cancer.

Ki-67、p53蛋白在乳腺癌组织的表达及临床意义

目的探讨Ki-67、p53蛋白在乳腺癌及癌旁组织中的表达及两者之间的关系和临床意义。方法应用免疫组化S-P法检测80例乳腺癌组织及其癌旁的正常乳腺组织中Ki-67、p53蛋白的表达,分析其与临床病理因素的关系及两者的相关性。结果80例乳腺癌组织中Ki-67蛋白阳性表达率为63.8%,高于癌旁乳腺组织8.8%(P<0.05),其表达与患者年龄、肿瘤大小、ER、PR表达无关,与肿瘤的分期、腋窝淋巴结转移、C-erbB-2表达相关。乳腺癌组织p53蛋白阳性表达率为68.8%,高于癌旁乳腺组织11.3%((P<0.05),其表达与年龄、肿瘤大小、分期、ER、PR表达无关,与腋窝淋巴结转移、C-erbB-2表达相关。Ki-67蛋白与p53蛋白表达之间无关。结论Ki-67、p53蛋白可通过抑制细胞凋亡,对乳腺癌发生和发展起重要作用,是判断乳腺癌生物学行为、预测转移趋势有价值的参考指标。

Ki-67和BRCA1在≤35岁乳腺癌组织中的表达及临床意义

目的研究Ki-67和BRCA1在年龄≤35岁乳腺癌组织中的表达与临床特点和病理学特征之间的关系。方法选取年龄≤35岁、临床资料完整的乳腺癌患者48例,应用免疫组织化学法测定癌组织中Ki-67和BRCA1的表达。对照组选取年龄>35岁乳腺癌患者35例,结合临床病理因素及乳腺癌分子分型进行相关分析。结果 (1)BRCA1在≤35岁和>35岁的乳腺癌患者中的表达率分别为79.2%、54.3%,差异有统计学意义(P<0.05)。Ki-67在≤35岁和>35岁的乳腺癌患者中的表达率差异无统计学意义(P>0.05);(2)在≤35岁乳腺癌患者组中,Ki-67的表达与BRCA1的表达呈负相关(r=-0.267,P<0.05)。(3)BRCA1及Ki-67表达在青年乳腺癌和中老年乳腺癌分子分型中分布差异均有统计学意义(P<0.05)。结论≤35岁乳腺癌患者肿瘤特征与中老年患者有差异,这种差异可能与≤35岁乳腺癌患者中Ki-67及BRCA1等生物学分子表达改变有关。

Ki-67抗原与子宫颈鳞癌新辅助化疗和放疗的敏感性研究

目的探讨Ki-67抗原与子宫颈鳞癌新辅助化疗(NACT)和放射治疗(RT)的敏感相关性。方法在新辅助化疗前后用免疫组化检测32例宫颈鳞癌组织Ki-67的表达。结果化疗前患者Ki-67的表达水平在不同年龄段组(<45岁和≥45岁)、不同临床期别组(Ⅰb～Ⅱa和Ⅱb～Ⅲb)和不同病理分级组(高中分化和低分化)之间相互比较差异有统计学意义(P=0.008、0.009、0.000)。化疗后患者Ki-67表达水平与化疗前比较明显下降,差异有统计学意义(P=0.000)。化疗前Ki-67表达程度不同,化疗和化放疗的疗效明显不同,化疗疗效高表达组与低表达组比较差异有统计学意义(P=0.033);化放疗疗效高表达组与低表达组比较差异无统计学意义(P=0.375);高表达组、低表达组对化疗和化放疗的疗效比较差异均有统计学意义(P=0.001、0.000)。结论子宫颈鳞癌治疗前Ki-67表达强度与化疗敏感性呈正相关;虽然存在Ki-67阳性表达强度愈强、NACT+RT的完全缓解率愈高的趋势,但未显示明显的放射敏感相关性;治疗前检测Ki-67表达可以为预测化疗和放疗敏感性提供客观依据,但Ki-67能否确定为预测子宫颈鳞癌新辅助化疗和放疗敏感性的可靠指标还需增加样本作进一步研究。

联合测定hTERT和Ki-67的表达对胶质瘤分级的评价

目的:探讨人脑胶质瘤恶性程度评定的客观标准。方法:采用组织微阵列技术,通过免疫组化的方法对60例不同级别人脑胶质瘤组织的hTERT和Ki-67表达水平进行测定,并与10例正常组织比较。结果:Ⅰ～Ⅱ级、Ⅲ级、Ⅳ级胶质瘤与对照组Ki-67表达阳性细胞标记指数分别为1.90±0.29、8.39±1.26、21.19±3.06与0.35±0.13,各组间差异有统计学意义,P<0.001;hTERT表达阳性细胞标记指数分别为5.30±0.65、28.63±3.59、53.62±4.68与0,各组间差异有统计学,P<0.01。hTERT与Ki-67表达阳性指数显著正相关,r=0.94,P<0.01,hTERT表达水平越高,Ki-67的表达也越高。结论:联合测定Ki-67和hTERT表达水平可以作为评价胶质瘤生物学行为、判断临床预后的重要参考指标。

乳腺癌18F-FDG PET/CT代谢参数与TNM分期及Ki-67表达的关系

目的探讨乳腺癌治疗前18F-FDG PET/CT显像中的3种代谢参数最大标准摄取值(maximum standardized uptake value,SUVmax)、肿瘤代谢体积(metabolic tumor volume,MTV)以及糖酵解总量(total lesion glycolysis,TLG)与肿瘤TNM分期及增殖细胞核抗原Ki-67的关系。方法回顾性分析52例乳腺癌患者治疗前的18F-FDG PET/CT显像。在影像工作站手动勾画病灶感兴趣区(region of interest,ROI),提取病灶SUVmax、MTV和TLG。采用Mann-Whitney秩和检验方法,分析3种PET/CT显像的代谢参数与TNM分期及Ki-67之间的关系。结果SUVmax、MTV和TLG这3种代谢参数在T分期T0~1组和T2~4组间比较,差异均具有统计学意义(均P<0.05)。MTV和TLG在N分期N0组和N1~3组间比较,差异均具有统计学意义(均P<0.05),而SUVmax在两组间比较,差异无统计学意义(P>0.05)。3种代谢参数在M分期M0组和M1组间比较,差异均无统计学意义(均P>0.05)。以20%作为Ki-67高低表达的阈值,3种代谢参数在Ki-67高表达组和低表达组间比较,差异均具有统计学意义(均P<0.05)。结论 18F-FDG PET/CT代谢参数MTV和TLG可以作为评估乳腺癌治疗前T、N分期及Ki-67指标的临床评价参数。

非小细胞肺癌组织中Skp2的表达及其与p27^(kip1)和Ki-67蛋白表达的关系

目的:探讨Skp2的表达在非小细胞肺癌(nonsmallcelllungcancer,NSCLC)发生发展中的作用,及其与p27kip1和Ki67蛋白表达的关系。方法:应用免疫组化SP法检测Skp2、p27kip1和Ki67三种蛋白在60例NSCLC和20例正常支气管黏膜上皮组织中的表达。结果:NSCLC组织中Skp2蛋白表达的阳性率为48.33%(29/60),显著高于正常支气管黏膜上皮组织中的表达,P=0.000。Skp2的表达与肿瘤的组织学类型、肿瘤细胞的分化程度、TNM分期、淋巴结转移和患者吸烟与否显著相关,P值分别为0.038、0.005、0.019、0.010和0.002,但与患者的年龄及性别无关,P值分别为0.833和0.281。NSCLC组织中Skp2表达与p27kip1表达呈显著负相关,P=0.001;而与Ki67表达呈显著正相关,P=0.027。结论:Skp2在NSCLC组织中表达是上调的,可能是通过作用于细胞周期调控蛋白p27kip1,加速了对p27kip1泛素化依赖的蛋白降解,使其表达及代谢发生异常,导致细胞周期失控并促进细胞异常增殖,从而参与了NSCLC的发生和发展。

鼻内翻性乳头状瘤中Ki-67的表达及与微血管密度的关系

1.1临床资料。收集1999年1月～2005年7月手术切除确诊为鼻内翻性乳头状瘤（nasal inverted papilloma,NIP）标本存档蜡块36例,男22例,女8例;鼻息肉（nasal polyps,NP）14例,男9例,女5例：鼻腔鳞状细胞癌（nasal squamous cell caminoma，NSCC）10例，男77例，女3例。

HIF-1α、Ki-67在NSCLC的表达及与放疗的关系

目的探讨非小细胞肺癌患者HIF-1α、Ki-67表达相关性与放疗疗效的关系。方法应用免疫组织化学法检测66例NSCLC组织标本中HIF-1α与Ki-67蛋白的表达,结合临床病理特征和随访资料进行分析。结果HIF-1α与Ki-67主要表达于细胞核,阳性表达率分别为56.1%和62.1%,两者存在正相关性(r=0.301,P=0.01)。HIF-1α表达与临床分期和淋巴结转移有关(P<0.05),而与组织学分级、病理分期及年龄、性别无明显关系(P>0.05)。HIF-1α阳性和阴性表达组近期缓解率,1、2、3年生存率分别为21.6%、56.8%、13.5%、0.0%和78.4%、90.9%、54.5%、32.7%;Ki-67阳性和阴性表达组近期缓解率1、2、3年生存率分别为61.0%、58.1%、14.0%、0.0%和28.0%、80.0%、66.7%、33.3%。结论 HIF-1α及Ki-67可作为放疗疗效的评价指标。

Ki-67、CyclinE、MDM2蛋白在尖锐湿疣组织中的表达

目的:探讨Ki- 6 7、cyclin E、MDM2蛋白在尖锐湿疣发病机制中的作用。方法:采用免疫组化SP法,检测了5 0例CA组织中Ki- 6 7、cyclin E、MDM2的原位表达,并以15例正常包皮组织及15例正常宫颈组织作对照。结果:Ki- 6 7的强阳性表达率、cyclin E阳性表达率及MDM2阳性表达率CA组均高于对照组(P<0 .0 5 )。CA皮损中Cyclin E、MDM2蛋白与Ki- 6 7呈正相关(r=0 .373,r=0 .393,P<0 .0 5 )。Cyclin E、MDM2蛋白的表达呈正相关(r=0 .4 4 2 ,P<0 .0 5 )。结论:Cylin E高表达及MDM2蛋白的异常表达可能与CA上皮的过度增殖有关,两者在CA的发病中可能起协同作用。

Ki-67在喉鳞状细胞癌的表达及临床病理学意义

目的:探讨Ki-67在喉鳞状细胞癌的表达及临床病理学意义。方法:采用免疫组化技术检测10例声带息肉、20例不典型增生病变和50例喉鳞癌组织中Ki-67的表达。结果:Ki-67阳性表达定位于细胞核,Ki-67在声带息肉组、不典型增生组和喉鳞癌组的阳性表达率分别为10%、25%和78%,统计学上有显著性差异,且Ki-67LI同肿瘤位置、组织学分级、临床分期及淋巴结是否转移相关。结论:Ki-67抗原在喉癌中具有显著的临床病理学意义,能用来反映喉癌的良恶性程度。

Expression of COX-2, PCNA, Ki-67 and p53 in gastrointestinal stromal tumors and its relationship with histopathological parameters

AIM: To investigate the expression of Cyclooxygenase-2 (COX-2), proliferating cell nuclear antigen (PCNA), Ki-67 and p53 in gastrointestinal stromal tumors (GISTs) and its relationship with histopathological parameters. METHODS: Twenty-five GISTs were examined by light microscopy and immunohistochemistry. c-kit, CD34, SMA, S-100 protein, COX-2, PCNA, Ki-67 and p53 were detected immunohistochemically and the relationship was evaluated among histopathologic parameters such as mitotic index (MI), tumor grade, tumor size, COX-2, PCNA, Ki-67 and p53. RESULTS: COX-2 protein expression was found in 19 of 25 (76%) of the tumors, and expression was noted in the cytoplasm of the tumor cells. p53 was significantly related to MI and tumor grade but no relationship was found between COX-2, proliferation markers and MI, tumor grade and tumor size. CONCLUSION: COX-2 is expressed in most GISTs and it may play an important role in the proliferation and progression of these tumors or a useful marker to identify GIST. Although immunohistochemical assessment of p53 can be used for distinguishing the risk groups of GISTs, tumor size and mitotic rate should be considered at the same time.

102例前列腺癌根治术后Ki-67的表达与预后价值分析

目的探讨Ki-67表达与临床病理特征之间的联系,并验证其在预测癌症特异性结果中的价值。方法使用组织微阵列(tissue microarray,TMA)对102例接受前列腺癌根治术治疗的有完整高质量Ki-67肿瘤数据的前列腺癌患者组织样本进行分析。免疫组织化学染色后,应用核定量算法(Aperio)分析每个癌核(3个癌核/例)肿瘤区域的Ki-67增殖指数(proliferative index,PI)。并评估Ki-67 PI是否与临床病理因素和无复发生存率(recurrence-free survival,RFS)相关,包括生化复发、转移或PC死亡(随访时间中位数为3年)。结果在102例前列腺癌中,Ki-67 PI显示肿瘤间变异高于肿瘤内变异(0. 68 vs 0. 39,P <0. 05)。Ki-67 PI与肿瘤分期(P <0. 01)、精囊浸润(P=0. 02)、包膜外扩散(P <0. 01)和Gleason评分(P <0. 01)相关。在多变量Cox比例危险模型中,Ki-67 PI作为连续变量与无复发、整体和疾病特异性生存率相关(HR=1. 04～1. 10,均P <0. 05)。高Ki-67评分(定义为≥5%)与RFS(HR=1. 47,P=0. 001)和总生存率(HR=2. 03,P=0. 030)较差相关。结论在前列腺根治术治疗的局限性前列腺癌患者中,较高Ki-67 PI与较高GS、癌症分期、SVI和ECE以及较高的复发率密切相关。

The predictive value of vascular endothelial growth factor and Ki-67 expression on neoadjuvant therapy in rectal cancer

Objective： To investigate the expressions of vascular endothelial growth factor （VEGF） and proliferating cell nuclear antigen （Ki-67） in patients with rectal adenocarcinoma and their associations with neoadjuvant therapy. Methods： The expressions of Ki-67 and VEGF in 32 cases of rectal adenocarcinoma, including both pretreatment tumor biopsies and postoperative specimen, were detected by immunohistochemistry using specific antibodies, and were correlated with clinicopathological factors. Results： The intensity of VEGF staining was significantly correlated with lymph nodal metastasis （P =0.033）, depth of tumor invasion （P =0.007） and tumor stage （P= 0.016）, but not with histological types, tumor sizes, patients＇ ages and genders （P 〉 0.05）. Low level of VEGF expression had significant correlation with the high sensitivity of response to neoadjuvant therapy （P = 0.016）. The transient increase of VEGF expression could be seen after neoadjuvant therapy （P = 0.035）. Ki-67 labeling index （Ki-67-LI） was significantly correlated with lymph node metastasis （P = 0.028）, but not correlated to tumor sizes, patients＇ ages and genders （P 〉 0.05）. Tumors with lower Ki-67-LI were more sensitive to neoadjuvant therapy （P = 0.032）. The Ki-67 level decreased after neoadjuvant therapy, but no statistical significance was found （P 〉 0.05）. Conclusion： Our results demonstrate that the expression of VEGF and Ki-67 in pretreatment rectal adenocarcinoma biopsies may be predictive of tumor response to neoadjuvant therapy.

Effect of Wuziyanzong pill on levels of sex hormones, and expressions of nuclear- associated antigen Ki-67 and androgen receptor in testes of young rats

OBJECTIVE: To evaluate the effects of Wuziyanzong pill on the levels of serum follicle stimulating hormone(FSH), luteinizing hormone(LH), testosterone(T) and the expressions of nuclear-associated antigen Ki67(Ki67), androgen receptor(AR) in testes of young rats.METHODS: Sixteen 20-day-old Wistar male rats were randomly divided into control and treatment group(n = 8). Rats in treatment group were administered Wuziyanzong pill by gavage; rats in control group administered the same volume of saline. After 10 days of treatment, the rats were killed, and then serum and testes were taken. The levels of FSH, LH and T were measured by radioimmunoassay(RIA). The histology of seminiferous tubule was observed by hematoxylin-eosin(HE) staining. The expression of Ki67 was detected by immunohistochemical assay(IHC). The m RNA level of Ki67, ARand CK-18 was detected with quantitative real-time polymerase chain reaction(Q-RT-PCR), the protein level of AR and CK-18 were tested by Western blot.RESULTS: Compared with control group, T level in treatment group increased significantly(P < 0.05).HE staining showed that both leydig cells and germ cells increased in treatment group. Expressions of Ki67 and AR became higher after treatment. There were no changes in CK-18 expression.CONCLUSION: Wuziyanzong pill can up-regulate AR level to promote germ cell proliferation and differentiation in young male rats.

Study of the expression of the gene encoding Ki-67 antigen in human pancreatic cancer using non-radioactive in situ hybridization and immunohistochemistry

bjective To investigate, at transcriptional and translational level in situ, whether the gene expression of the Ki 67 protein in pancreatic carcinoma specimens is altered to get insight of the gene structure and function. Methods Forty pancreatic cancer, 5 normal pancreatic and 4 chronic pancreatitis tissues were used in this experiment. A 435 bp cDNA fragment located in codon 2, exon 13 of Ki 67 antigen gene was amplified by the polymerase chain reaction (PCR). The DIG labeled cRNA probes were transcribed using a commercial DIG RNA labeling kit. Localization of the Ki 67 protein and the specific mRNA was performed by combining immunohistochemistry (ICH) with DIG labeled in situ hybridization (ISH). Results Successful localization of the Ki 67 protein mRNA in pancreatic tissue sections, routinely formalin fixed and paraffin embedded , was first accomplished in this experiment. Analysis of the Ki 67 mRNA transcription in 17 pancreatic cancer specimens with Ki 67 ICH labeling index >20% revealed stronger mRNA signals in poorly differentiated specimens with Ki 67 index >50% than in well differentiated cases with the ICH labeling index of 20% 50%. A high expression of both the mRNA and the protein was observed in pancreatic adenocarcinomas with poor differentiation. Conclusions This is the first study in which the abnormal overexpression of the gene encoding Ki 67 protein was detected not only at the protein level, but also at the mRNA level in pancreatic tumours. The abnormal overexpression of the Ki 67 protein might be correlated with the central part, exon 13, of the gene.

Ki-67在胃癌组织中的表达及其临床意义

目的 检测Ki-67在不同胃癌组织中的表达,分析Ki-67的表达与临床病理特征的相关性及其对预后的影响.方法 应用免疫组化二步法,检测60例胃癌患者术后标本中Ki-67的表达情况,并结合临床病理特征及随访资料进行综合分析.采用四格表Chi-square检验和Fisher确切概率法分析Ki-67在不同组织类型胃癌中的表达差异;采用Spearman进行等级相关分析,分析Ki-67分别表达及双表达与临床病理特征的相关性,采用Kaplan-Meier法和Log-rank检验进行生存期分析,分析Ki-67表达对胃癌术后患者无病生存期及总生存期的影响.结果 ⑴胃癌组织中Ki-67阳性表达率为65%（31/60）,在不同组织类型的胃癌中表达差异无统计学意义（P＞0.05）.⑵Ki-67的表达与浸润深度、临床分期有关（rs＝0.269,0.265,P＜0.05）,Ki-67表达阳性的患者无病生存期及中位生存期均明显低于表达阴性者（P＝0.011,0.021）.结论 Ki-67的表达与肿瘤浸润深度及TNM分期相关,Ki-67阳性表达者生存期劣于阴性表达者.

肺癌组织中胸苷酸合酶与细胞增生核抗原蛋白的表达

目的研究胸苷酸合酶(TS)、细胞增生核抗原(Ki-67)在非小细胞肺癌(NSCLC)癌组织中的表达及临床意义;观察TS、Ki-67在NSCLC癌组织表达水平的相关性。方法应用免疫组化SP染色法检测了51例不同临床特征的NSCLC癌组织标本的TS和Ki-67的表达水平。结果TS与Ki-67蛋白总阳性率分别为68.6%(35/51)、72.5%(37/51),在不同组织类型癌组织的表达差异均无显著性;TS与癌组织分化程度差异有显著性(P<0.05),与TNM分期、淋巴结转移差异无显著性(P>0.05);Ki-67与分化、TNM分期、淋巴结转移差异有显著性(P<0.05);TS与Ki-67两者之间的表达水平呈显著正相关(r=0.84),TS各组别的Ki-67表达具有显著差异性。结论TS、Ki-67是判断NSCLC分化及增殖程度有价值的指标。

MAGE-A3与NSCLC胸腔积液治疗效果的相关性研究

目的探讨MAGE-A3在非小细胞肺癌(non-small cell lung cancer,NSCLC)患者胸腔积液中的表达与临床疗效的关系,为个体化治疗提供指导。方法回顾性分析62例NSCLC(48例肺腺癌、14例鳞癌)患者胸腔积液中MAGE-A3和Ki-67抗原表达水平,比较其与临床疗效的相关性。结果 MAGE-A3、Ki-67在NSCLC患者胸腔积液的表达水平与临床疗效呈负相关,且两者表达水平呈正相关(均P<0.05)。其中,胸腔积液MAGE-A3表达阳性患者较表达阴性者临床疗效差(P<0.05)。MAGE-A3表达水平与年龄、性别、胸腔积液量、癌症类型、KPS评分无相关性(P>0.05)。结论 MAGE-A3表达可能在NSCLC侵袭转移中发挥重要作用,为胸腔积液疗效评估及个体化治疗策略制定提供依据。