99mTc-MIBI显像在原发性甲状旁腺功能亢进定位诊断中的应用价值

目的:评价99mTc-甲氧基异丁基异腈(99mTc-MIBI)显像在原发性甲状旁腺功能亢进症(PHPT) 定位诊断中的临床应用价值。方法:回顾性分析57例疑似PHPT患者99mTc-MIBI显像、血清PTH测定及B超检查结果,以术后病理为金标准,对99mTc-MIBI双时相显像、血清PTH测定及B超检查三种诊断方法的灵敏度、特异度、准确度进行比较和评价。结果:术后病理证实,57例患者99mTc-MIBI显像与血清PTH测定两种诊断方法比较,灵敏度、特异度、准确度分别为91.49%（80.85%）、50.00%（80.00%）、84.42%（80.70%）;99mTc-MIBI显像与B超检查两种诊断方法比较,灵敏度、特异度、准确度分别为91.49%（76.60%）、50.00%（70.00%）、84.21%（75.44%）。结论:99mTc-MIBI显像对PHPT定位诊断的灵敏度、准确度都高于血清PTH测定及B超检查结果,但特异性较低。99mTc-MIBI显像联合血清PTH测定及B超可以提高PHPT的诊断准确性,具有更高的临床应用价值。     

Comparison of idarubicin and daunorubicin regarding intracellular uptake, induction of apoptosis, and resistance

Anthracycline antibiotics are widely used as anticancer agents. Idarubicin (4-demethoxydaunorubicin; Ida), a semisynthetic derivative of daunorubicin (Dnr) is more potent than the parent compound in vitro and in vivo. The equitoxic dose of Ida in patients is about one-fourth of that of Dnr. We compared these drugs regarding cytotoxicity, apoptosis induction, and resistance mechanisms in human leukaemic cell lines. Cytotoxicity was studied by means of the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide assay and drug-induced apoptosis by means of the Annexin V-fluorescein isothiocyanate method at similar intracellular concentrations (extracellular concentrations of 0.35 microM for Ida and 1 microM for Dnr). Ida was at least twice as potent as Dnr in MOLT-4, HL60, CEM, and K562 cell lines. It took 8 h for Ida to induce approximately 20% apoptosis, but at least 22 h for Dnr to reach 20% apoptosis at identical intracellular concentration. Ida induces a faster and higher apoptosis rate compared with Dnr. The human chronic myelogenous leukaemia cell line (K562) was selected for resistance to Dnr and Ida with and without verapamil (Ver). Continuous incubation with Dnr, but not with Ida, led to an increased mdr1 gene expression as assessed by real-time PCR. The development of mdr1 gene expression in Dnr-resistant cells could be reversed by the presence of Ver. Ver also reversed the cytotoxicity to Dnr, but not to Ida, in K562/Dnr cells. The results show that Ida is more effective than Dnr in inducing apoptosis and that there are differences in resistance mechanisms between the drugs.     

乳腺肿瘤摄取99Tcm-甲氧基异丁基异睛与耐多药蛋白表达的关系

目的研究乳腺肿瘤摄取99Tcm-甲氧基异丁基异睛 (99Tcm-MIBI)与耐多药蛋白表达水平间的相关性.方法 30例经病理证实的原发性浸润性导管癌患者,均行99Tcm-MIBI早期显像和延迟显像.对显像阳性的肿块计算感兴趣区与对侧正常相应部位的放射性计数比值,以早期摄取比值(EUR)和延迟摄取比值(DUR)计算滞留指数(RI).以免疫组织化学法检测手术切除标本肿瘤组织的P-糖蛋白(P-gp)和多药耐药相关蛋白(MRP)的表达水平.数据分别进行配对t检验、Pearson相关分析及偏相关分析.结果 P-gp的平均表达水平为0.1183±0.0700,MRP的平均表达水平为0.1195±0.0522.P-gp和MRP的表达水平在RI≥0和RI＜0的组中,差异有显著性.P-gp的表达与RI(r=-0.919,P=0.001)、DUR(r=-0.675,P=0.001)和MRP(r=0.549,P=0.002)有相关性,与EUR(r=-0.097,P=0.610)无相关性.MRP的表达与RI(r=-0.547,P=0.002)有相关性,与EUR(r=0.292,P=0.117)、DUR(r=-0.173,P=0.361)无相关性.P-gp与MRP的表达有相关性(r=0.549,P=0.001).偏相关分析则显示,P-gp的表达与RI(r=-0.8847,P=0.001)有显著相关性,MRP的表达与RI(r=-0.1296,P=0.512)无相关性.结论 99Tcm-MIBI乳腺显像与P-gp的表达呈负相关关系,与MRP未见有相关性.     

99Tcm-MIBI脑显像对脑胶质瘤恶性程度的诊断及对预后的预测

背景与目的:判断胶质细胞瘤恶性程度对制订治疗方案、评估预后具有重要意义.传统的影像学诊断方法如CT、MRI对胶质瘤的分级仅限于提供解剖信息,有时难以明确诊断.立体定向活检为有创性检查,仅能反映局部病理变化,而且标本采集误差会使肿瘤分级不准确.18F-FDG PET显像可直接反映肿瘤的葡萄糖代谢率,被认为是最有价值的方法,但因费用昂贵目前尚不能广泛应用.本研究通过99Tcm-MIBI脑肿瘤显像,测定不同病理类型胶质瘤对99Tcm-MIBI的摄取,以探讨其对胶质瘤恶性程度诊断及预测预后的价值.方法:回顾性分析进行99Tcm-MIBI脑显像的52例胶质瘤患者及15例对照者的临床资料.计算99Tcm-MIBI脑SPECT诊断脑胶质瘤的敏感性、特异性、准确率.采用t检验比较病灶T/N比值在不同病理类型组间的差异.随访患者生存率,将患者按T/N比值在不同区间分组,平均生存期(MST)在不同T/N比值组间的比较采用t检验.结果:99Tcm-MIBI显像发现44例阳性,2例假阳性.诊断敏感性为84.6%(44/52),特异性为86.7%(13/15),准确率为85.1%.早期显像及延迟显像中,Ⅰ级星形细胞瘤与Ⅱ级之间以及室管膜瘤与少突胶质细胞瘤之间,T/N比值未见显著性差异.其它任意两组进行比较,随胶质瘤恶性程度增高,T/N比值显著提高(P＜0.001).T/N比值越高,患者MST越短.T/N比值1.2～2.0组MST显著高于T/N比值大于或等于4.1组(t=5.412,P＜0.001)和T/N比值为3.1～4.0组(t=4.418,P＜0.001);T/N比值2.1～3.0组MST亦显著高于大于或等于4.1组(t=3.382,P＜0.005)和T/N比值为3.1～4.0组(t=2.389,P＜0.05).T/N比值1.2～2.0组与2.1～3.0组之间以及3.1～4.0与大于或等于4.1组之间MST无显著性差异(P＞0.05).结论:99Tcm-MIBI脑显像对胶质瘤的诊断和恶性程度的鉴别有一定价值.密切监测残留病灶摄取强度的变化,可判断肿瘤的生物学行为,为预测预后提供帮助.     

Overexpression of the mdr1 gene in blast cells from patients with acute myelocytic leukemia is associated with decreased anthracycline accumulation that can be restored by cyclosporin-A

Typical multi-drug resistance (MDR) in human and animal cell lines is caused by overactivity of a unidirectional drug efflux pump. This pump is composed of a 170-kDa transmembrane glycoprotein (P-glycoprotein) that is encoded by the so-called mdr1 gene. The functionally relevant characteristic of MDR cells is a defect in drug accumulation that can be restored by agents which inhibit the P-glycoprotein pump. The purpose of our study was to find out whether P-glycoprotein inhibitors could increase the daunorubicin (DNR) accumulation in acute myelocytic leukemia (AML) cells, overexpressing the mdr1 gene. Using dot blot analysis with an mdr1-specific cDNA probe, we identified leukemic cell samples, obtained from chemotherapy-resistant AML patients, that had relatively high levels of mdr1 expression. These leukemic cells showed a reduced ability to accumulate DNR in vitro, as quantitated by flow cytometry. Addition of cyclosporin-A (Cy-A), a drug known to inhibit the P-glycoprotein pump, to the incubation medium resulted in an increase (up to 60%) in steady-state drug uptake by the leukemic cells. The degree of Cy-A-induced increase in drug accumulation in the leukemic cells correlated approximately with the level of overexpression of the mdr1 gene. Our data indicate that Cy-A is a good candidate for combination chemotherapy with cytotoxic drugs in clinical trials, aimed at the treatment of drug resistance in AML.     

p53和c—myc异常表达与膀胱癌多药耐药性

为探讨ｐ５３和ｃ－ｍｙｃ基因异常表达与膀胱癌多药耐药性（ＭＤＲ）的关系，应用ＡＢＣ免疫组织化学方法检测６７例膀胱癌癌细胞中ｐ５３和ｃ－ｍｙｃ的表达产物及多药耐药基因（ｍｄｒ－１）产物Ｐ－ｇｐ。结果显示ｐ５３突变蛋白蓄积的膀胱癌中Ｐ－ｇｐ阳性表达率９２．３％；ｐ５３阴性者Ｐ－ｇｐ阳性率为３６．６％。Ｐ－ｇｐ表达与ｐ５３突变蛋白蓄积呈显著正相关（ｒ＝０．６４，Ｐ＜０．０５）；而ｃ－ｍｙｃ和ｍｄｒ－１的表达无明显相关。提示：ｐ５３异常表达可增加ｍｄｒ－１基因的表达，使膀胱癌细胞获得ＭＤＲ表型     

吲哚美辛对A549/DDP细胞摄取99Tcm-MIBI的影响及其逆转耐药性的机制

目的：探讨吲哚美辛对A549/DDP摄取99Tcm的影响及其逆转耐药性的机制.方法：常规方法培养A549/DDP细胞,首先行99Tcm-MIBI摄取实验,然后用DDP和吲哚美辛干预A549/DDP细胞30min后计算99Tcm-MIBI的摄取率,再分别用RT-PCR,Westen-blot检测mdrl基因,Pg-P蛋白的表达.结果：A549/DDP细胞摄取率在90min时达到高峰（0.83%）,吲哚美辛干预后99Tcm-MIBI摄取比干预前增加,达到3.79%.RT-PCR,Westen-blot结果显示吲哚美辛抑制了mdrl基因和Pg-P蛋白的表达.结论：吲哚美辛能提高A549/DDP细胞99Tcm摄取率并通过抑制mdrl和Pg-P的表达逆转肺癌耐药.99Tcm放射性摄取率可作为评价肿瘤耐药的一种方法.     

Association of p53 expression with proliferative activity and poor-prognosis in patients with squamous-cell lung carcinomas

Tumor tissue of 65 patients with squamous cell lung carcinoma was analyzed for p53 expression using immunohistochemistry (DO-1) and for proliferative activity using flow cytometry. Of the 65 cases, 17 cases (26%) showed positive staining for p53, whereas 48 cases (74%) showed no expression. The median survival time for patients with p53-negative tumors was 100 weeks and for patients with p53-positive tumors 30 weeks (rank-sum test, p=0.03; log-rank test, p=0.14). The median survival time for patients with high proliferative activity (proportion of SG(2)M-phase cells >22%) was one year and for patients with low proliferative activity (proportion of SG(2)M-phase cells less than or equal to 22%) over 6 years (rank-sum test, p=0.04; log-rank test, p=0.01). There exists a trend that p53-positive squamous cell lung carcinoma had a higher proportion of SG(2)M-phase cells than p53-negative tumors. Multivariate analysis found independent prognostic significance for proliferative activity and stage but not for p53.     

siRNA对SGC7901/VCR细胞mdr1基因沉默效果的影响因素分析

目的分析siRNA沉默人胃癌SGC7901/VCR细胞mdr1基因效果的相关因素。方法设计并体外转录合成4条靶向mdr1的siRNA(mdr1si326、mdr1si1513、mdr1si2631和mdr1si3071),转染SGC7901/VCR细胞,用RTPCR和免疫印迹检测mdr1mRNA和Pgp的表达、流式细胞仪检测细胞内阿霉素的蓄积和MTT法检测细胞对阿霉素的敏感性,综合这4方面结果评价4条siRNA的沉默效果情况;用分子生物学软分析siRNA沉默效果的影响因素。结果沉默效果最好的mdr1si326和较好的mdr1si2631靶序列编码Pgp的跨膜区而且自身无茎和袢;沉默效果较差的mdr1si3071和最差的mdr1si1513靶序列编码Pgp的胞内区,前者自身成茎和成袢。沉默效果最好的mdr1si326和较好的mdr1si2631靶序列在靶位点和靶位点外间有较少的碱基配对和氢键。siRNA的沉默效果与siRNA3’5’端3个碱基中的A/U数量、N1和N19、GC含量间无规律可循。结论siRNA沉默SGC7901/VCR细胞mdr1的效果与靶序列的结构关系密切。     

Further evidence that altered p16/CDKN2 gene expression is associated with lymph node metastasis in squamous cell carcinoma of the esophagus

Esophageal squamous cell carcinoma (ESCC) has high malignant potential with a poor outcome. Lymph node metastasis is the most useful indicator for predicting the outcome of ESCC. The p16/MTS1/CDKN2 gene and the cyclin D1/PRAD-1 gene cooperatively regulate CDK4-mediated phosphorylation of RB protein in the cell cycle. We immunohistochemically detected p16, cyclin D1, and RB expressions in both primary lesions and metastatic lymph nodes in ESCC. Among the 50 ESCC primary lesions, 24 (48%) were positive for p16, while 26 (52%) were negative for p16. Sixteen (32%) were p16-positive, 34 (68%) were p16-negative among the 50 ESCC metastatic lymph nodes. Eight cases (16%) were p16-positive in primary lesion and p16-negative in lymph node, however, no cases that was p16-negative in the primary tumor exhibited p16-positivity in metastatic lymph nodes (p < 0.0001). Seventeen (34%) of the 50 ESCC primary lesions were cyclin D1-positive, while 33 (66%) were cyclin D1-negative. Twenty-four (48%) were cyclin D1-positive, 26 (52%) were cyclin D1-negative among the 50 metastatic lymph nodes. Five cases (10%) were cyclin D1-positive in primary lesion and cyclin D1-negative in lymph node, and 12 cases (24%) were cyclin D1-negative in primary lesion and cyclin D1-positive in lymph nodes. Nine cases (18%) were RB-negative in 50 primary lesions, and the rate of loss of RB expression in metastatic lymph nodes was not markedly higher than in primary lesions. Thirty-nine (78%) of 50 primary lesions and 46 (92%) of 50 metastatic lymph nodes had altered expression of at least one of the three G1 control genes. Tumor cell with disruption of these cell cycle regulators can get a growth advantage and metastatic potential during tumor progression, especially p16/CDKN2 alterations may be associated with lymph node metastasis in ESCC. These results also suggest that tumor cells in metastatic lymph nodes may have more aggressive proliferation and higher malignant potential than tumor cells in primary lesions.     

The significance of CD34 and TdT determinations in patients with untreated de novo acute myeloid leukemia.

The results of intensive chemotherapy given to 247 adults at the University of Maryland Cancer Center with previously untreated de novo acute myeloid leukemia (AML) were reviewed with respect to expression of terminal deoxynucleotidyl transferase (TdT) and CD34. Of the 228 patients with data for TdT, 32 (14%) had > 5% of the leukemia cells positive by an immunofluorescence assay. The median age of the TdT-positive patients was approximately 10 years less than the TdT-negative patients (50 versus 60 years). Patients with TdT-positive AML had similar median survival (12 versus 10.5 months) and complete remission (CR) rates (53 versus 59%), but a greater frequency of long-term complete responders (60 of complete remitters versus 20%, p = 0.08) than TdT-negative patients. Of 126 patients tested, 59% were CD34-negative (< 20% reactivity with leukemia cells). These 74 patients (median age 60 years) had a greater CR rate (71 versus 48%, p = 0.008) than the 52 CD34-positive patients (median age 60 years), and improved survival (p = 0.013 by Wilcoxon) although there was no difference in the duration of CR between the CD34-positive and negative groups. Of CD34-positive patients 12/52 remain in continuous CR, and 16/74 CD34-negative patients remain in continuous CR. None of eight patients strongly positive for CD34 (> 70% reactivity) remain disease-free. Positivity for TdT or CD34 was associated with less differentiated AML. Of CD34-positive patients, 44% had FAB M0/M1 morphology versus 13% of CD34-negative patients (p = 0.0001); similarly, 47% of TdT-positive patients were FAB M0/ML1 versus 25% of TdT-negative patients (p = 0.01). Of seven patients with FAB M4E0, five were CD34-positive. Of the 12 CD34-positive survivors, four had FAB M4E0. Thus CD34 expression predicts for CR rate and overall survival in adults with AML. TdT expression does not significantly affect overall outcome but may be associated with longer CR durations.     

Prognostic value of beta1 integrin expression in metastatic melanoma

The expression of integrin-type cell adhesion receptors is frequently changed in malignant transformation. Despite their important role in cancer cell behaviour, the value of integrins as prognostic markers is mostly unknown. We have examined the expression of beta1 integrins in 38 metastatic melanomas obtained from 27 patients treated with combined chemoimmunotherapy. On the basis of beta1 integrin expression, the melanoma samples were divided into two groups: beta1-negative tumours (<10% beta1 integrin immunostained cells) and beta1-positive tumours (with > or = 10% positive cells). Patients with beta1-positive tumours (n = 15) had significantly longer disease-free survival (median 38 versus 7 months, P < 0.0001) and overall survival (median 70 versus 23 months, P = 0.0001) evaluated after the diagnosis of primary disease compared with patients with beta1-negative metastases (n = 11). Moreover, the survival of the patients with beta1-positive tumours after the initiation of chemoimmunotherapy was significantly prolonged (median 18 versus 9 months, P = 0.017). The independent nature of beta1 integrin expression as a significant prognostic factor for survival after therapy was confirmed using Cox's multivariate analysis (P = 0.014). Our results indicate that the expression of beta1 integrins might have some major tumour growth regulatory role and can be used as a predictor for prognosis in patients with metastatic melanoma.     

Patients with ERCC1-negative locally advanced esophageal cancers may benefit from preoperative chemoradiotherapy

PURPOSE: To assess the significance of excision repair cross-complementation group 1 (ERCC1) expression as a predictive marker, we analyzed the effects of preoperative chemoradiotherapy on survival relative to ERCC1 status in patients with locally advanced operable esophageal cancer. EXPERIMENTAL DESIGN: Paraffin-embedded pretreatment tumor specimens, collected by endoscopic biopsy from patients treated with surgery alone or with preoperative chemoradiotherapy followed by surgery, were immunohistochemically assayed for ERCC1 expression. RESULTS: Of the 175 patients, 152 biopsy specimens were available for immunohistochemical analysis. Based on a median ERCC1 expression score of 1, we divided the samples into ERCC1-positive (score >1; 71 patients, 47%) and ERCC1-negative (score 相似文献   

核酶对人肺腺癌细胞多药耐药的逆转

目的 探讨针对多药耐药基因（ｍｄｒｌ）的核酶对肺腺癌细胞多药耐药的逆转作用。方法以ｍｄｒｌ ｍＲＮＡ ８８０位点为靶位点,通过脂质介导,将抗ｍｄｒｌ核酶表达质粒（ｐＨ β Ａｐｒ－ｌ ｎｅｏ／ｍｄｒｌ－Ｒｂ）导入肺腺癌耐药细胞株Ａ５４９／Ｒ。分别采用ＲＴ－ＰＣＲ、流式细胞仪检测术、罗丹明聚集及ＭＴＴ方法,观察细胞的ｍｄｒｌ ｍＲＮＡ、Ｐ糖蛋白（Ｐ－ｇｌｙｃｏｐｒｏｔｅｉｎ,Ｐｇｐ）表达和对阿霉素敏     

ERCC1 expression as a prognostic marker in N2(+) nonsmall-cell lung cancer patients treated with platinum-based neoadjuvant concurrent chemoradiotherapy

BACKGROUND.: Excision repair cross-complementation Group 1 (ERCC1) overexpression is associated with resistance to cisplatin-based chemotherapy in patients with nonsmall-cell lung cancer (NSCLC). A preliminary study also suggested that ERCC1 expression is associated with radioresistance in lung cancer cells. The aim of this study was to evaluate the clinical implications of ERCC1 expression in stage IIIA N2-positive NSCLC patients treated with platinum-based neoadjuvant concurrent chemoradiotherapy (CCRT) followed by surgery. METHODS.: Sixty-eight patients with mediastinoscopy-proven N2-positive NSCLC were enrolled between August 1997 and September 2003. ERCC1 expression was assessed by immunohistochemistry from pretreatment mediastinoscopic biopsy specimens. RESULTS.: ERCC1 expression was positive in 31 of 68 specimens (46%). Among 14 patients who obtained pathologic complete response, 6 were positive for ERCC1 expression and 8 were negative (P = .818). On univariate analysis, with median follow-up of 61.8 months (range, 34.3-108.8 months), progression-free survival was 15.9 months for ERCC1-positive and 29.5 months for ERCC1-negative patients (P = .062), and there was a statistically significant difference in overall survival between ERCC1-negative tumors and ERCC1-positive tumors (89.2 vs 26.0 months, P = .014). On multivariate analysis, ERCC1 negativity (P = .041) and achieving mediastinal nodal clearance (downstage to pathological N0 or N1) after neoadjuvant CCRT followed by surgery (P = .005) were significant independent prognostic factors for the prolongation of survival. CONCLUSIONS.: These results suggest that N2-positive NSCLC patients with ERCC1 negative tumors show a survival benefit from neoadjuvant CCRT with a platinum-containing regimen. Cancer 2008. (c) 2008 American Cancer Society.     

Expression of the mdr3 gene in prolymphocytic leukemia: association with cyclosporin-A-induced increase in drug accumulation

Typical multidrug resistance in human and animal cell lines is caused by overactivity of an unidirectional transmembrane drug efflux pump, encoded by the MDR genes, called mdr genes in mice and humans and pgp genes in hamsters. In humans, two mdr genes, mdr1 and mdr3, with approximately 80% nucleotide homology, have been identified. There is increasing evidence that overexpression of the mdr1 gene plays a role in resistance to anticancer agents in specific tumor types. However, currently no data are available on a possible role for mdr3 in drug resistance. Here we report high levels of expression of mdr3 gene sequences in leukemic cells from 6 out of 6 patients with prolymphocytic leukemia (PLL). No mdr1 expression was detected in 5 out of 6 of these samples, whereas a low level of mdr1 expression was found in a sample from one PLL patient in the course of transformation to non-Hodgkin's lymphoma. Except for this patient, all other PLL cases studied had not received prior chemotherapy. In vitro drug uptake studies showed that daunorubicin accumulation in PLL cells was increased by cyclosporin A. Since cyclosporin A is an inhibitor of the mdr1-encoded P-glycoprotein drug pump, these data suggest that in PLL cells mdr3 also codes for a drug efflux pump. Our findings could partly explain the primary refractoriness of PLL to chemotherapy.