非小细胞肺癌患者淋巴结、外周血及骨髓微转移分子诊断在肺癌外科治疗中的作用

目的 探讨检测肺癌患者区域淋巴结、外周血和骨髓中微转移在肺癌外科治疗中的临床意义,以及区域淋巴结、外周血和骨髓肺癌微转移三者之间的相关性。方法 应用巢式RT－PCR技术,对29例肺癌患者和11例肺良性病变患者的淋巴结、外周血和骨髓中CK19基因mRNA表达进行联合检测。结果 本实验建立的巢式RT－PCR技术的敏感性可达到10^-6。术前10例患者检测到外周血微转移,6例患者检测到骨髓肺癌微转移,101枚纵隔淋巴结中55枚检测到肺癌微转移。肺癌患者淋巴结、外周血和骨髓中微转移阳性检出率分别为54．5％、34．5％和20．7％,三者之间存在显著正相关（P＜0．05）,外周血和骨髓微转移与肺癌组织学类型、细胞分化程度及P－TNM分期均存在密切关系（P＜0．05或P＜0．01）。结论 RT－PCR法是一种特异性,敏感性均较高的肿瘤微转移检测方法;检测CK19 mRNA表达应用于肺癌微转移的分子诊断有助于选择外科手术适应证,并具有广阔的临床应用前景。     

RT-PCR法检测MUC1 mRNA诊断肺癌纵隔淋巴结隐匿转移

目的：探讨对常规病理检查漏诊的肺癌纵隔淋巴结转移病灶的诊断方法。方法：应用逆转录聚合酶链反应法（RT－PCR），检测pN0.1期非小细胞肺癌患者（NSCLC）纵隔淋巴结中MUC1基因mRNA的表达。结果：5枚肺良性疾病的局部淋巴结无MUC1基因mRNA表达，5枚经病理检查证实有淋巴结转移癌的NSCLC纵隔淋巴结中均检测中MUC1mRNA表达，实验组19例患者的78例枚纵隔淋巴结中有6枚检测到MUC1mRNA表达，从而诊断为纵隔淋巴结隐匿转移。结论：应用RT－PCR法检测纵隔淋巴结中MUC1基因mRNA的表达可以提高临床对肺癌纵隔淋巴结转移诊断的准确性。     

肺癌患者淋巴结微转移MUC1表达的探讨

目的:探讨非小细胞肺癌(NSCLC)患者淋巴结微转移MUC1表达用于分子诊断特异性和敏感性及临床意义.方法:应用逆转录聚合酶链反应(RT-PCR)技术,对31例肺癌患者119枚和10例肺良性病变患者35枚淋巴结MUC1 mRNA表达进行检测.结果:巢式RT-PCR技术的敏感性达10-6.RT-PCR检测的119枚肺癌患者淋巴结中65枚存在MUC1 mRNA表达,阳性率为54.6%(65/119);病理学方法检测出41枚淋巴结存在癌转移,阳性率为34.5%(41/119).肺良性病变患者35枚淋巴结中MUC1 mRNA表达均为阴性.淋巴结微转移与肺癌组织学类型、细胞分化程度及pTNM分期均存在密切关系(P＜0.05).结论:RT-PCR是一种特异性、敏感性均较高的肿瘤微转移检测方法,MUC1 mRNA可能是检测肺癌微转移的一个有价值的指标,为制定治疗方案和评估预后提供重要的参考依据.     

肺癌患者外周血和骨髓中MUC1 mRNA表达的临床意义

目的:检测非小细胞肺癌(non smal celllungcancer,NSCLC)外周血和骨髓中粘蛋 白1(mucin1,MUC1)mRNA的表达。方法:应用 RT PCR技术,对31例肺癌患者、10例肺良性病 变患者和8名健康人外周血及骨髓中MUC1基 因mRNA表达进行检测。结果:31例肺癌患者 中10例检测到外周血,7例检测到骨髓中有 MUC1mRNA表达,10例肺良性病变患者和8名 健康人中未检测到MUC1mRNA表达。肺癌患 者外周血及骨髓中MUC1mRNA表达分别为 32.3%(10/31)和22.6%(7/31),两者之间存在 正相关关系,P<0.05;外周血和骨髓中MUC1 mRNA的表达与肺癌组织学类型、细胞分化程度 及pTNM分期均密切相关,P<0.05。结论:肺癌 患者外周血和骨髓中存在常规方法检测不出的 肿瘤细胞;应用巢式RT PCR法可检测到肺癌患 者外周血和骨髓中MUC1mRNA表达,可为制 定治疗方案和评估预后提供重要参考依据。     

检测CK19 mRNA和MUC1 mRNA诊断非小细胞肺癌淋巴结微转移

目的 探讨非小细胞肺癌(NSCLC)淋巴结微转移的基因诊断方法，并分析CK19mRNA、MUC1mRNA作为肺癌微转移检测分子标志物的可行性。方法应用巢式逆转录聚合酶链反应(nestedRT-PCR)，对31例NSCLC的119枚淋巴结中的粘蛋白1(MUC1)mRNA、角蛋白19(CK19)mRNA表达情况进行检测；对照组为10例肺良性病变患者的35枚淋巴结。结果 31例NSCLC患者的119枚淋巴结中，66枚(55.5％)淋巴结存在CK19mRNA阳性表达，65枚(54.5％)存在MUC1mRNA阳性表达；肺良性病变患者35枚淋巴结中CK19mRNA和MUC1mRNA表达均为阴性，与肺癌组比较均有显著性差异。结论 MUC1、CK19基因均可作为RT-PcR法检测NSCLC患者淋巴结微转移的分子标志物，两者联合检测可能有助于早期诊断肺癌转移。     

肺癌患者三种微转移标志物临床意义的比较研究

目的评价肺癌患者外周血LUNX mRNA 、CK19 mRNA、CEA mRNA诊断微转移的特异性、敏感性,探索肺癌患者微转移的早期诊断.方法以肺部良性疾病30例、健康人10例的外周血为对照,以LUNX mRNA 、CK19 mRNA、CEA mRNA的表达为指标,采用逆转录聚合酶链反应(RT-PCR)检测48例肺癌患者外周血、手术切除区域44个淋巴结的微转移,并根据淋巴结病理切片、临床分期、随访过程中复发转移率评价微转移的临床意义.结果①LUNX mRNA、CK19 mRNA、CEA mRNA在肺癌组织的表达率均为100%(35/35).②48例肺癌患者外周血中LUNX mRNA阳性30例(62.5%),CK19 mRNA阳性24例(50.0%),CEA mRNA阳性32例(66.7%);44枚肺癌患者手术切除的区域淋巴结,LUNX mRNA阳性16枚(36.4%),CK19 mRNA阳性12枚(27.3%),CEA mRNA阳性18枚(40.9%).③30例肺部良性疾病患者外周血中,CK19 mRNA表达阳性2例(6.7%),LUNX mRNA或CEA mRNA表达均阴性;10例健康人外周血和11枚取自肺良性疾病患者的淋巴结,3个指标检测结果均为阴性.④44枚肺癌区域淋巴结中,普通病理组织学检出6枚(13.6%)有癌转移,阳性率明显低于RT-PCR检测结果(P<0.05).⑤3种微转移指标的阳性率均随TNM分期的增加而升高(P=0.01).⑥随访发现外周血微转移阳性患者的复发率高于阴性患者.结论 LUNX mRNA、CK19 mRNA 、CEA mRNA有可能成为监测肺癌微转移的分子标志.     

CK19mRNA,MUC1mRNA,LUNX mRNA在非小细胞肺癌微转移中的表达及临床意义

目的:检测非小细胞肺癌(NSCLC)和非肿瘤患者骨髓和外周血中肿瘤特异性标志物细胞角蛋白19(CK19)、黏蛋白1(MUC1)和肺特异性蛋白X(LUMX) mRNA表达情况,探讨微转移与肿瘤的病理类型、分期的关系.方法:选取肺癌患者30例,健康对照组15例,应用逆转录聚合酶链反应(RT-PCR)方法检测CK19、MUC1和LUNX三项标志物.结果:患者骨髓和外周血中CK19 mRNA阳性率分别为36.7%和30%,其表达状况与患者的肿瘤病理类型、吸烟史无明显关系,而与肿瘤的病理分期关系密切;MUC1 mRNA 阳性率分别为6.7% 、6.7%,其表达与肿瘤的分期、病理类型、患者的吸烟史均无显著关系,与骨髓及外周血中微转移的关系尚待确定;LUNX mRNA阳性率分别为16.7%和13.3%,可以作为指示NSCLC微转移的敏感特异的指标.结论:三种检测标志物中CK19 mRNA对微转移的诊断敏感性较高,但存在假阳性的可能;LUNX mRNA作为一个新的检测指标对微转移的诊断有一定的作用,但特异性和敏感性不高;MUC1 mRNA与骨髓及外周血中微转移的关系尚待确定.     

食管癌淋巴结隐匿性转移的基因诊断

目的：探讨食管癌淋巴结隐匿性转移的基因诊断方法。方法：应用逆转录聚合酶链反应法（RT－PCR），检测阴性对照组及阳性对照组淋巴结MUC1基因mRNA表达，计算特异度及灵敏度；检测食管鳞癌手术后病理诊断阴性的区域淋巴结（pN0)中MUC1基因mRNA表达，诊断淋巴结隐匿性转移。结果：阴性对照组30个标本（食管良性疾病区域淋巴结）均无MUC1基因mRNA表达，特异度为100%；阳性对照组30个标本（食管鳞癌病理证实有转移癌的区域淋巴结）中25枚检测到MUC1 mRNA的表达，灵敏度为83％。实验组30例pN0食管癌的87枚区域淋巴结中，其中8例病人的9枚淋巴结检测到MUC1mRNA表达（检出率10.3%)。结论：应用RT－PCR法检测食管癌区域淋巴结中MUC1基因mRNA的表达可以诊断淋巴结隐匿性转移。     

肺癌淋巴结隐匿性转移的基因诊断及预后研究

目的 探讨非小细胞肺癌（NSCLC）纵隔淋巴结隐匿性转移的基因诊断方法,并评价其对预后的意义。方法 应用逆转录聚合酶链反应（RT－PCR）技术,对37例pN0 NSCLC（Ⅰa-Ⅱb期）的168枚纵隔淋巴结标本中的黏蛋白1（MUC1）基因mRNA表达情况进行检测。对照组为同期经手术治疗的患者,其中阴性对照淋巴结30枚（取自14例肺良性病变）,阳性对照标本30个（15例NSCLC的肿瘤标本和病理诊断为转移癌的NSCLC纵隔淋巴结15枚）。通过x^2检验,比较预后差别。结果 阴性对照组均无MUC1 mRNA表达,特异度1005;阳性对照组30个标本中,26个有MUC1 mRNA表达,灵敏度86.7%。实验组中12例患者的16个标本有MUC1 mRNA表达,占9.5%,其TNM分期上调为Ⅲa。MUC1 mRNA表达阴性组3年生存率（88.0%）高于MUC1 mRNA阳性组（58.3%,P＜0.05）。结论 应用RT－PCR法检测MUC1 mRNA的表达可诊断肺癌纵隔淋巴结隐匿性转移;纵隔淋巴结隐匿性转移可能与pN0肺癌预后不良有关。     

Irinotecan and radiation in vitro and in vivo

Loss of chromosome sequences at 13q14 (Rbl) and 17p13 (p53) associated with squamous cell carcinoma of head and neck (SCCHN) was evaluated in 12 recurrent tumors and 51 primary tumors from 63 patients. The incidence of LOH at 17p13 was 19 of 50 (38%) tumors, and at 13q14 was 21 of 57 (37%). LOH affecting Rbl and/or p53 was observed in 30 of 63 (48%) SCCHN. Coincident LOH at Rbl and p53 was detected in 10 of 46 (22%) tumors. There were nine cases in which primary and metastatic tumors were obtained from the same patient. Of these, seven were informative and five of these (71%) manifested LOH at p53 in both primary and metastatic sites. Examination of Rbl in these same tumors showed LOH in six of the nine metastases, and of these six, only three revealed LOH in the primary tumor. LOH at p53 or Rbl alone showed no correlation with clinical outcome. However, tumors that manifested LOH at both loci were associated with poorer patient outcome and poorer histological differentiation.     

Daunorubicin and daunorubicinol pharmacokinetics in plasma and tissues in the rat

Recent evidence suggests that 13-hydroxy metabolites of anthracyclines may contribute to cardiotoxicity. This study was designed to determine the pharmacokinetics of daunorubicin and the 13-hydroxy metabolite daunorubicinol in plasma and tissues, including the heart. Fisher 344 rats received 5 mg kg^–1 daunorubicin i.v. by bolus injection. Rats were killed at selected intervals for up to 1 week after daunorubicin administration for determination of concentrations of daunorubicin and daunorubicinol in the plasma, heart, liver, kidney, lung, and skeletal muscle. Peak concentrations of daunorubicin were higher than those of daunorubicinol in the plasma (133±7 versus 36±2 ng ml^–1;P<0.05), heart (15.2±1.4 versus 3.4±0.4 g g^–1;P<0.05), and other tissues. However, the apparent elimination half-life of daunorubicinol was longer than that of daunorubicin in most tissues, including the plasma (23.1 versus 14.5 h) and heart (38.5 versus 19.3 h). In addition, areas under the concentration/time curves (AUC) obtained for daunorubicinol exceeded those found for daunorubicin in almost all tissues, with the ratios being 1.9 in plasma and 1.7 in the heart. The ratio of daunorubicinol to daunorubicin concentrations increased dramatically with time from <1 at up to 1 h to 87 at 168 h in cardiac tissue. Thus, following daunorubicin injection, cumulative exposure (AUC) to daunorubicinol was greater than that to daunorubicin in the plasma and heart. If daunorubicinol has equivalent or greater potency than daunorubicin in causing impairment of myocardial function, it may make an important contribution to the pathogenesis of cardiotoxicity.     

TCP and NTCP in preclinical and clinical research in Europe

European research in radiation oncology has a long and successful tradition. The aim of this research is to increase the therapeutic window of radiotherapy by increasing the tumor control probability (TCP) and/or by decreasing the normal tissue complication probability (NTCP). This paper summarizes the basic radiobiological concept underlying treatment optimization by TCP-NTCP data and discusses some of the limitations of currently used models. These are controversial in many aspects and cannot be recommended for clinical routine practice but should rather be considered as a research tool.     

Foscan and foslip based photodynamic therapy in osteosarcoma in vitro and in intratibial mouse models

Current osteosarcoma therapies cause severe treatment‐related side effects and chemoresistance, and have low success rates. Consequently, alternative treatment options are urgently needed. Photodynamic therapy (PDT) is a minimally invasive, local therapy with proven clinical efficacy for a variety of tumor types. PDT is cytotoxic, provokes anti‐vascular effects and stimulates tumor cell targeting mechanisms of the immune system and, consequently, has potential as a novel therapy for osteosarcoma patients. This study investigated the uptake and the dark‐ and phototoxicity and cytotoxic mechanisms of the photosensitizer (PS) 5,10,15,20‐tetrakis(meta‐hydroxyphenyl) chlorine (mTHPC, Foscan) and a liposomal mTHPC formulation (Foslip) in the human 143B and a mouse K7M2‐derived osteosaroma cell line (K7M2L2) in vitro. Second, the tumor‐ and metastasis‐suppressive efficacies of mTHPC formulations based PDT and associated mechanisms in intratibial, metastasizing osteosarcoma mouse models (143B/SCID and syngeneic K7M2L2/BALB/c) were studied. The uptake of Foscan and Foslip in vitro was time‐ and dose‐dependent and resulted in mTHPC and light dose‐dependent phototoxicity associated with apoptosis. In vivo, the uptake of both i.v. administered mTHPC formulations was higher in tumor than in healthy control tissue. PDT caused significant (Foscan p < 0.05, Foslip p < 0.001) tumor growth inhibition in both models. A significant (Foscan p < 0.001, Foslip p < 0.001) immune system‐dependent suppression of lung metastasis was only observed in the K7M2L2/BALB/c model and was associated with a marked infiltration of T‐lymphocytes at the primary tumor site. In conclusion, mTHPC‐based PDT is effective in clinically relevant experimental osteosarcoma and suppresses lung metastasis in immunocompetent mice with beneficial effects of the liposomal mTHPC formulation Foslip.     

Hyperthermia and platinum complexes: Time between treatments and synergy in vitro and in vivo

To investigate the greatest therapeutic efficacy, we investigated the effect of scheduling on the cytotoxic interaction between hyperthermia and seven different platinum complexes in vitro and in vivo using the FSaII murine fibrosarcoma cells. Hyperthermia treatment (43°C, 1 h) was administered at various times relative to exposure of the cells to the IC₉₀ (at 37°C, 1 h) of each platinum complex. Greater-than-additive killing of FSaII cells was obtained with cis-diamminedichloroplatinum (II) (CDDP) and hyperthermia when the drug and heat exposure were overlapping or simultaneous. The same cell killing effect with carboplatin and hyperthermia resulted from heat exposure up to 5 h prior to, simultaneous with, or immediately after the drug exposure. D-Tetraplatin and K₂PtCl₄ were synergistic with hyperthermia only if the drug and heat exposure were simultaneous. PtCl₄(Nile Blue)₂ and hyperthermia produced greater-than-additive cell killing if the heat and drug exposure occurred in immediate sequence, simultaneously, or with drug exposure up to 5 h prior to heat exposure. PtCl₄(Rh-123)₂ and hyperthermia produced greater-than-additive cell killing if the drug and heat occurred in immediate sequence, overlapping, or simultaneously. PtCl₄(Fast Black)₂ and hyperthermia were additive over a wide range of scheduling from heat exposure 2 h prior to 5 h after drug exposure. When animals bearing FSaIIC tumours were treated with single doses of CDDP (10 mg/kg), carboplatin/PtCl₄(Nile Blue)₂ (50 mg/kg), PtCl₄(Rh-123)₂/PtCl₄(Fast Black)₂ (100 mg/kg) under various combined schedules with hyperthermia treatment (43°C, 30 min), similar cytotoxicity patterns were observed. To administer hyperthermia at a time when the drug concentration in the tumour tissue is at peak level, careful scheduling of systemically administered anticancer drugs with hyperthermia is needed. Modelling studies can identify the stringency/flexibility of drug/heat scheduling to achieve synergistic tumour cell killing.     

Fibronectin and integrins in invasion and metastasis

Summary The adhesive glycoprotein fibronectin and integrin receptors appear to play important roles in the progression of metastatic disease. Fibronectin is a multifunctional extracellular glycoprotein that has at least two independent cell adhesion regions with different receptor specificities. The cell adhesive region in the central portion of fibronectin is comprised of at least two minimal amino acid sequences - an Arg-Gly-Asp (RGD) sequence and a Pro-His-Ser-Arg-Asn (PHSRN) sequence - which function in synergy. Another cell adhesive region is located near the carboxy-terminus in the alternatively spliced IIICS module. The critical minimal sequences for this region are Leu-Asp-Val (LDV) and Arg-Glu-Asp-Val (REDV) which function in an additive rather than synergistic fashion. Integrins are heterodimeric, transmembrane cell adhesion receptors for fibronectin and other extracellular matrix molecules. Several different integrins bind to fibronectin. The ₅₁ fibronectin-specific integrin binds to the central RGD/PHSRN site. The ₄₁ integrin binds to the IIICS site. Fibronectin-integrin interactions are important in tumor cell migration, invasion, and metastasis. In addition to promoting cell adhesion to the extracellular matrix, these proteins may also function in chemotaxis and control of proliferation. Peptide and antibody inhibitors of fibronectin and integrin functions have been shown to be effective inhibitors of metastasis, and are potentially important reagents for the study and control of cancer.     

Racial Differences in Oral and Pharyngeal Cancer Treatment and Survival in Florida

OBJECTIVE: This study examined racial differences in treatment and survival for blacks and whites in Florida diagnosed with oral or pharyngeal cancer. METHODS: Data for 21,481 malignancies of the oral cavity or pharynx diagnosed from 1988 to 1998 were derived from the Florida Cancer Data System. Type of cancer treatment was compared by race, stratified by anatomic site and summary stage at diagnosis. Kaplan-Meier survival curves were used to compare survival rates and Cox regression models were used to estimate hazard ratios for race. Covariates included age, sex, census tract income, and treatment. RESULTS: Stratifying by tumor site and stage, blacks consistently had poorer survival rates than whites. Across tumor stages, blacks with oral cavity cancer were consistently more likely than whites to have received only radiotherapy and less likely to have received cancer-directed surgery. Trends were similar for pharyngeal cancer, although statistically significant only for regional stages. Across site and stage, blacks consistently had elevated hazard ratios (range: 1.20-1.53) relative to whites. CONCLUSIONS: In Florida, there were racial differences in patient treatment for oral or pharyngeal cancer. Blacks had lower survival rates than whites, but differences in treatment did not entirely account for racial disparities in survival.     

Synergistic anti-cancer effects of immunotoxin and cyclosporin in vitro and in vivo

Background:

The clinical use of immunotoxins (ITs) has been hampered by hepatotoxicity, and the induction of a strong human-anti-IT response. The human-anti-IT response results in neutralisation of the immunoconjugates, rendering repetitive treatment inefficacious.

Methods:

We evaluated the combination of cyclosporin A (CsA) with various Pseudomonas exotoxin A-based ITs in human breast, cervical, and prostate cancer cell lines measured by protein synthesis, cell viability, and TUNEL assay. Furthermore, expression of essential proteins were analysed by western blot. We used cervical cancer model in nude rats to evaluate the anti-metastatic effect of the combination. The anti-immunogenic response by the CsA treatment was investigated in immunocompetent rats.

Results:

The combination of CsA with ITs caused remarkable synergistic cytotoxicity, in several cancer cell lines, characterised by protein synthesis inhibition, decreased cell viability, and an increased apoptotic index. Furthermore, the combination strongly inhibited formation of metastases in a cervical cancer model in nude rats with a statistically significant increase in median survival time of the combination-treated animals, as compared with those receiving a suboptimal dose of IT alone. Notably, we found in immunocompetent rats that the anti-IT immunoresponse elicited by repeated administration of IT was efficiently abrogated by CsA; notably the antibody responds towards the highly immunogenic PE was shown to be prevented.

Conclusion:

The combination of ITs and CsA might constitute a significant improvement in the clinical potential of systemic IT treatment of cancer patients.