EGF及其受体flt对荷瘤鼠喉癌细胞增殖及凋亡的影响

目的　探讨VEGF及其受体flt对荷瘤鼠喉癌细胞增殖及凋亡的影响。方法　首先建立荷瘤 (HEP 2喉癌细胞 )裸鼠动物模型 ,将抗VEGF抗体及抗VEGF受体flt抗体分别注射荷瘤鼠体内 ,通过活体标记BrdU的方法 ,观察抗VEGF抗体及抗flt抗体对喉癌HEP 2细胞增殖的影响 ;用Tunel法观察对喉癌细胞凋亡的影响。结果　抗VEGF及抗flt抗体组HEP 2细胞BrdU标记阳性率显著小于对照组 (P <0 .0 5 ) ,细胞凋亡指数则显著高于对照组 (P <0 .0 5 )。结论　以抗体中和VEGF及封闭VEGF的结合位点均可抑制喉癌细胞的增殖 ,促进喉癌细胞凋亡。VEGF与喉癌细胞的生长密切相关。     

VEGF及其受体flt对荷瘤鼠喉癌细胞增殖的影响

目的;观察血管内皮生长因子（ＶＥＧＦ）对喉癌细胞增殖动力学的影响。方法：首先建立荷瘤（ＨＥＰ－２喉癌细胞）裸鼠动物模型,将抗ＶＥＧＦ抗体及抗ＶＥＧＦ受体ｆｌｔ抗体分别注射荷瘤鼠体内,通过活栓标记ＢｒｄＵ的方法,观察抗ＶＥＧＦ抗体及ｆｌｔ抗体对喉癌ＨＥＰ－２细胞增殖的影响。结果：与对照组相比较,抗ＶＥＧＦ抗体治疗组ＨＥＰ－２细胞ｂｒｄＵ标记阳性率显著降低;     

整合素αvβ3对喉鳞癌细胞增殖和侵袭力影响的离体研究

目的探讨整合素αvβ3在喉鳞癌细胞增殖和侵袭性中的作用,为以整合素αvβ3为靶点治疗喉鳞癌提供实验依据。方法采用MTT比色法和PCNA免疫组化,观察不同浓度的整合素αvβ3抗体对体外培养的Hep-2喉鳞癌细胞增殖的影响;建立Boyden小室细胞侵袭模型,观察不同浓度的整合素αvβ3抗体对体外培养的Hep-2喉鳞癌细胞侵袭能力的影响。结果整合素αvβ3抗体对体外培养的HEP-2喉鳞癌细胞的增殖有明显抑制作用,其作用表现出明显的量效关系（P〈0.05）;整合素αvβ3抗体显著降低体外培养的HEP-2喉鳞癌细胞的侵袭能力,其作用亦表现出明显的量效关系（P〈0.05）。结论整合素αvβ3对喉鳞癌细胞的恶性增殖和侵袭性生长具有促进作用,为抗整合素αvβ3治疗喉鳞癌提供了实验依据。     

抗VEGF抗体及抗KDR抗体对裸鼠胃癌细胞增殖和凋亡的影响

目的探讨抗VEGF抗体及抗KDR抗体对胃癌细胞增殖和凋亡的影响。方法建立胃癌原位种植模型;7天后注射抗VEGF抗体、抗KDR抗体及二者联合注射,连续给药8周,每周2次,第10周末处死动物,用免疫组化法、RT-PCR法检测移植瘤组织中VEGF蛋白和mRNA表达情况;FCM检测肿瘤细胞增殖及凋亡情况。结果与对照组相比,抗VEGF抗体和抗KDR抗体组肿瘤组织中VEGF蛋白的表达降低,而VEGFmRNA表达无显著变化;肿瘤细胞发生不同程度的凋亡,凋亡率明显增高,G0/G1期细胞明显增多,S期细胞明显减少,细胞增殖指数降低;两者合用时作用更为明显。结论通过中和VEGF抗体和封闭KDR受体抑制血管形成,可能为胃癌的抗血管生成治疗提供依据。     

端粒酶反义RNA 对人喉癌细胞系Hep-2 生长的影晌

 目的　探讨抗端粒酶在喉癌细胞治疗中的意义。方法　用脂质体介导法将 pBBS2 12 hTR(反义端粒酶RNA真核表达载体 )转染入人喉癌Hep 2细胞系中 ,采用TRAP法测定端粒酶活性 ,细胞动力学检测 ,电镜下观察等方法研究其对Hep 2细胞的端粒酶活性、细胞增殖及细胞凋亡影响。 结果　端粒酶反义RNA能显著抑制细胞的端粒酶活性 ,抑制细胞增殖并促进其凋亡。结论　以端粒酶反义RNA抑制端粒酶活性是治疗喉癌的潜在途径。     

耐药喉癌Hep-2/VCR细胞ATP7B表达与顺铂耐药的相关性

目的 检测copper transporting P-type adenosine triphosphatase(ATP7B)在人喉癌耐药细胞株中的表达,并研究其与喉癌细胞顺铂耐药的关系.方法 人喉鳞癌Hep-2细胞和Hep-2/长春新碱(vincristine,VCR)耐药细胞分别给予顺铂作用24小时和48小时,应用细胞增殖/毒性检查试剂盒(cell counting kit-8,CCK8)细胞活力试验、末端脱氧核苷酸转移酶介导的dUTP缺口末端标记测定法[terminal dexynueleotidyl transferase (TdT)-mediated dUTPnick end labeling,TUNEL]细胞凋亡实验分别检测细胞增殖和凋亡.分别用免疫荧光染色和实时RT-PCR检测ATP7B蛋白和mRNA在细胞中的表达.结果 50 μmol/L和l00μmol/L的顺铂作用24小时后,Hep-2细胞活力下降为71.0％,而Hep-2/VCR细胞活力无下降,差异有统计学意义(P＜0.01).50 μmol/L顺铂能够诱导Hep-2和Hep-2/VCR细胞表达ATP7B.Hep-2细胞在作用后3小时表达增强,6小时达到峰值,12小时下调(P＜0.05);而Hep-2/VCR细胞在3小时表达增强,6小时达到峰值(P＜0.05),持续高表达24小时无明显变化.结论 喉癌耐药细胞Hep-2/VCR的ATP7B蛋白的高表达与喉癌细胞的顺铂耐药有关.     

海芋粗提物对人肝癌细胞的细胞毒和凋亡诱导作用

目的:探讨海芋粗提物对人肝癌细胞的细胞毒和凋亡诱导作用及其作用机制.方法:用海芋粗提物处理人正常肝细胞(L02)及肝癌细胞(SMMC-7721),用细胞增殖试验(MTT法)、克隆形成试验、EdU掺入试验研究海芋粗提物对人正常肝细胞和肝癌细胞的增殖抑制作用;吖啶橙/溴乙啶(AO/EB)染色检测细胞凋亡;流式细胞术检测海芋粗提物对细胞周期的影响;RT-PCR检测细胞周期及凋亡相关基因PPARγ、Cyclin D1、Rb、P21、Bax、Bcl-2基因mRNA表达.结果:用不同浓度海芋粗提物处理后,肿瘤细胞增殖活性降低(P＜0.05),呈时间-剂量-效应关系;克隆形成下降(P＜0.05);EdU标记指数降低(P＜0.05);AO/EB染色凋亡细胞增多(P＜0.01);细胞周期分布改变,凋亡指数显著增加(P＜0.01),G0/G1期细胞增加(P＜0.05),S期和G2/M期细胞减少(P＜0.05);RT-PCR结果显示,海芋粗提物(400μg/ml)处理后,PPARγ表达增加46.0%(P＜0.01)、Cyclin D1降低43.7% (P＜0.01)、Rb增加283.3%(P＜0.01)、Bax增加77.1% (P＜0.01)、Bcl-2降低24.8% (P＜0.05).结论:海芋粗提物对肝癌细胞具有细胞毒和诱导凋亡作用,其作用机理可能与激活PPARγ,上调Rb、Bax基因表达,下调Cyclin D1、Bcl-2基因表达有关.     

Notch2在喉鳞状细胞癌中的表达及意义

目的 探讨Notch2在喉癌组织及体外培养的喉鳞癌细胞Hep-2中的表达及与喉癌患者临床病理资料之间的关系.方法 应用量子点超敏免疫荧光染色法检测95例喉癌组织标本中Notch2蛋白的表达,同时选取31例声带息肉标本作为对照组,结合患者临床病理资料进行分析;制作细胞爬片并进行量子点(QDs)免疫荧光染色检测Notch2蛋白在喉鳞癌细胞Hep-2中的表达.结果 95例喉癌组织中Notch2阳性表达率为92.6％ (88/95),其阳性率与声带息肉中的阳性率(32.3％)相比差异均有统计学意义(P＜0.01),其表达主要定位于细胞核.Notch2在喉癌组织中的表达与临床分期显著相关(P＜0.05):临床分期越高,表达率越高.Notch2在喉癌组织细胞核中表达阳性与T分级、临床分期、病理分级和淋巴结转移显著相关(P＜0.05).喉癌细胞Hep-2中Notch2表达阳性,其表达主要定位于细胞质和胞膜中.结论 Notch2在喉癌的发生发展中起着重要作用,其可能成为治疗喉癌的分子靶点.     

地塞米松对喉鳞状细胞癌血管内皮生长因子-C及血管内皮生长因子受体-3表达的影响

目的 探讨地塞米松(DEX)对人类喉鳞状细胞癌原代细胞血管内皮生长因子-C(VEGF-C)、VEGF受体-3(VEGFR-3)基因表达的影响.方法 12例喉癌患者喉癌原代细胞培养成功后分两组,孵育至对数生长期,一组继续培养,另一组给予DEX 0.1μmol/L,37℃,5% CO2孵箱孵育24 h,用RT-PCR检测其VEGF-C、VEGFR-3在两组中的基因表达情况.结果 VEGF-C、VEGFR-3在喉癌原代培养细胞表达阳性率分别为83%、58%,DEX干预组喉癌原代培养细胞表达阳性率分别25%、8%,VEGF-C、VEGFR-3在喉癌细胞中的表达显著高于DEX作用组(P＜0.05).结论 DEX作用于人类喉鳞状细胞癌原代细胞,可导致VEGF-C、VEGFR-3的表达降低.可能为临床喉癌化疗及喉癌喉梗阻应用DEX提供理论依据.     

雌激素和孕激素对人宫颈癌细胞体外生长影响的研究

目的:探讨雌二醇(E2)和孕酮(P)对人宫颈癌细胞(Hela细胞及Siha细胞)体外增殖及凋亡的影响.方法:体外培养人宫颈癌Hela细胞及Siha细胞,采用四甲基偶氮唑蓝(MTT)比色法检测两种激素对Hela细胞及Siha细胞增殖的影响;流式细胞仪(FCM)检测细胞生长周期和凋亡率的影响;免疫组化方法测定其雌激素受体(ER) 和孕激素受体(PR)的表达.结果:E2对Hela细胞有明显促生长作用,P<0.05. P和E2+P对Hela细胞生长有抑制作用,P<0.05;对Siha细胞无明显作用,P>0.05.FCM显示,E2组Hela细胞S期细胞比例上升,凋亡率下降,P和E2+P组Hela细胞S期细胞比例下降,凋亡率上升,E2、P和E2+P组对Siha细胞的生长周期及凋亡率并无显著影响,P>0.05.E2和P对人宫颈癌细胞ER和PR的表达无显著影响.结论:E2对宫颈癌Hela细胞具有促进生长抗凋亡作用,而P抑制其生长诱导凋亡,P能有效抵消E2对Hela细胞生长的刺激作用.E2和P对Siha细胞生长无明显影响.     

Experience in the treatment of synchronous and metachronous carcinoma of the oesophagus and the head and neck

BACKGROUND AND OBJECTIVES: Treatment of multiple primary squamous cell carcinomas of the head and neck and oesophagus is controversial. The poor prognosis of these 2 types of carcinoma taken individually and their anatomic proximity complicate the therapeutic strategy and limit the treatment choices for each location. METHODS: From 1986 to 1998, 43 patients received curative treatment for multiple synchronous (n = 30) or metachronous (n = 13) primary neoplasms of the oesophagus and head and neck. For synchronous cancers, the therapeutic strategy consisted of first curing the head and neck cancer and then planning oesophagectomy according to the type of head and neck cancer therapy. RESULTS: Ten total oesopharyngolaryngectomies and 33 subtotal oesophagectomies were performed. The postoperative mortality rate was 9.3% (4/43). The rate of anastomotic leakage was 30% (13/43), and all such leaks were cervical. Pulmonary infection occurred in 19% of cases (8/43). A past history of cervical radiation therapy or cervicotomy did not appear to be a significant risk factor for anastomotic leakage or pulmonary complications. Oesophagectomy did not affect the functional results in the 31 patients whose larynx could be preserved. CONCLUSIONS: Oesophagectomy after head and neck cancer treatment is possible with a low mortality rate and acceptable morbidity.     

Clinicopathological study of carcinomas of the lip and the mucosa of the upper and lower lips

BACKGROUND: Lip carcinomas are rare oral tumors, and there have been few reports of lip carcinoma in Japan. METHODS: Of 914 patients with oral carcinomas treated between January 1980 and December 1998, 12 (1.3%) had lip carcinoma and 5 (0.5%) had lip mucosal carcinoma. We investigated the clinicopathological features of these 17 patients. RESULTS: Of the 12 patients with carcinoma of the lip, 10 had squamous cell carcinomas (9, external lower lip; 1 commissures) and 2 had mucoepidermoid carcinomas (external upper lip). Of the 5 patients with lip mucosal carcinoma, 3 had squamous cell carcinomas (2, mucosa of the lower lip; 1, mucosa of the upper lip), 1 had mucoepidermoid carcinoma (mucosa of the lower lip), and 1 had acinic cell carcinoma (mucosa of the lower lip). Of the 12 patients with lip carcinoma, 9 were classified as stage I, 2 as stage II, and 1 as stage III; all 5 of the patients with lip mucosal carcinoma were stage I. Five patients with lip carcinoma were treated by resection, 5 by a combination of resection and reconstruction, and 2 by radiotherapy alone. All patients with lip mucosal carcinoma were treated by resection. After the initial therapy, 3 patients without neck dissection had regional recurrences and received delayed neck dissection, and 2 died with neck regional recurrence after dissection. The 5-year cumulative survival rates of the patients with lip carcinoma and those with lip mucosal carcinoma were 82.5% and 80.0%, respectively. CONCLUSION: We suggest that early-stage carcinomas of the lip and of the mucosa of the upper and lower lips are frequent, and we found that the outcome of these patients was excellent. However, an aggressive therapeutic approach to the lip carcinoma patient with cervical metastasis appears warranted, in an attempt to improve locoregional control and ultimate survival.     

The relation between the aging of the steroid generating system and the geneses of cancers of the stomach, the breast and the uterine cervix

The purpose of the present study is to test the validity of the steroid carcinogenesis hypothesis in humans by investigating the problem whether or not a cancer-specific change of the hormonal milieu emerges at a specified stage of life where the growth rate of cancer risk is at its zenith. A case-control study of 14 urinary steroid excretions was conducted for each of 3 human neoplasias. The identification and the size (in parenthesis) of the population units used in this study were,given as follows: a) the male gastric cancer group (421); b) the male control group (104); c) the female breast cancer group (245); d) the cervical cancer group (345); e) the female control group (127). Two kinds of steroid parameters were employed for the statistical analysis of hormonal data: a) the logarithm of a steroid excretion figure (mu g/day), as expressed by log x; b) the logarithm of a relative weight of a given steroid to tetrahydrocortisol, as expressed by log x/THF. The case-control difference for each parameter was expressed in terms of a t-value of Student's t-test. The steroid deviation profile was prepared for each neoplasia and for each of the log x data set and the log x/THF data set. The results obtained are as follows: a) the 2 steroid parameters (log x and log x/THF) for each of 14 urinary steroids were both subject to change with the progress of host age. The rate of age-dependent change was different for each steroid parameter and for each population unit. b) The above differential age dependency of the steroid parameters gave rise to a continual transition of the steroid deviation profile in the course of aging. c) The hormonal traits of male gastric cancer, female breast cancer and cervical cancer were described each as a complex of androgen depression and glucocorticoid stimulation (male gastric cancer), a sequential emergence of premenopausal progestin depression and postmenopausal predominance of glucocorticoid over androgen (female breast cancer), and a complex of androgen-glucocorticoid depression over progestin (cervical cancer). d) The emergence of the above cancer-specific steroid disorders chronologically coincided with the quasiexponential growth phase of cancer risk (and slow growth phase of cancer risk in postmenopausal breast cancer). e) The usefulness of the log x/THF type deviation profile for the assessment of the hormonal milieu of the host was verified by both theoretical approach to the problem and its application to the real data of a case-control study. f) The age dependent decline of androgens was generally much faster in their progressions than that of glucocorticoids - a finding to suggest the possibility that the production of a cancer-specific steroid deviation profile might have taken the form of the stress shift of Hans Selye, since both phenomena share depletion of gonadal steroids relative to glucocorticoid in common. The etiological relevancy of the 3 cancer-specific steroid changes to the geneses of 3 cancers:was discussed in the light of the experimental pathology studies in our laboratory as well as in other laboratories.     

Effect of freezing the helix and the rim or edge of the human and pig ear

We have studied the effect of increasing freeze times on the normal pig's ear and on a variety of lesions of the human ear. The clinical and laboratory data suggest that cartilage necrosis secondary to cryosurgery is a dose-related phenomenon and is uncommon with the freeze times used in clinical practice. Cryosurgery is an effective and cosmetically acceptable treatment for superficial skin lesions of the ear.     

Degree of manifestation of therapeutic pathomorphosis and the nature of the change in the estrogen and progesterone receptors after the radiation and chemotherapy of breast cancer

Estradiol and progesterone receptor levels were measured in 130 patients with stage III breast tumors before treatment and following preoperative radiation or chemotherapy. The data were evaluated versus the morphologic features of posttreatment pathomorphosis of tumor. Standard fractionated radiation (total dose of 70 Gy) was followed by pronounced postradiation pathomorphosis and a decrease in the level and incidence of steroid receptors in 72.7-87.5%. The essentially unchanged receptor profile of tumor following large-fraction (total dose-20 Gy) irradiation as well as presence of estradiol and progesterone receptors in the originally receptor-negative neoplasms after chemotherapy were matched by a slight degree of pathomorphosis.     

Urbanization and the incidence of abnormalities of squamous and glandular epithelium of the cervix

BACKGROUND: The large data bases of the Dutch cervical screening program can be exploited to establish the relation between urbanization and the incidence of abnormalities of the squamous and glandular epithelium, including mild or greater changes of the squamous and glandular epithelium of the cervix. METHODS: Six cytology laboratories in the context of the Dutch cervical screening program screened over 190,000 cervical smears. Urbanization (place of residence) data were derived from postal codes. All smears were coded with the Dutch national coding system, the Dutch national classification system KOPAC, in which squamous abnormalities are coded S4-S9, and glandular cell changes are coded G4-G9. From the scores per 1000 screened women, the relative risk (RR) of living in a large city compared with living in rural areas was calculated. To investigate a trend in incidence in relation to urbanization, the Schaafsma method was used. RESULTS: Of the smears with positive cytology, mild squamous dysplasia (S4) had the highest incidence per 1000 screened women (4.32), and the lowest incidence was found for adenocarcinoma (in situ; G7/G9; RR, 0.07). The RR for urban women ranged from 1.73 for moderate squamous dysplasia (S5) to 7.55 for adenocarcinoma (in situ; G7/G9). For smears with positive cytology for both squamous and glandular abnormalities, the Schaafsma method indicated a significant positive trend. CONCLUSIONS: The incidence of squamous and glandular abnormalities are maximal in women who live in a large city, which, in The Netherlands, is where there also is a population at high risk for human papillomavirus and bacterial vaginosis.     

Patient-reported outcomes: Assessment and current perspectives of the guidelines of the Food and Drug Administration and the reflection paper of the European Medicines Agency

AimsPatient-reported outcomes (PROs) have recently gained greater credibility with regulatory bodies aiming to standardise their use and interpretation in RCTs, thereby supporting medicinal product submissions. For this reason, the United States (US) Food and Drug Administration (FDA) and the European Medicines Agency (EMEA) have released guidelines. This review paper provides an overview of the current perspectives and views on these guidelines.MethodTo evaluate the FDA and EMEA PRO guidelines, 47 expert responses to the FDA guidance were qualitatively reviewed. Two reviewers independently extracted data from these letters and checked these responses to warrant consistency and agreement in the evaluation process. A PubMed literature review was systematically examined to obtain supporting evidence or related articles for both the guidance documents.ResultsGenerally, there is agreement between regulatory authorities and the research community on the contents of the FDA and EMEA PRO draft guidance. However, disagreements exist on significant philosophical topics (e.g. the FDA focuses more on conceptual models and symptoms than the EMEA) and design topics (e.g. the FDA is more restrictive on issues of recall bias, blinding of oncology trials and degrees of psychometric validation than researchers and the EMEA). This could influence the approval of PRO claims.ConclusionPRO guidance from the EMEA and FDA has been valuable, and has raised the profile and active debate of PROs in oncology. However, our review of the current opinion shows that there are controversial aspects of the guidance. Consequently, greater latitude should be given to how the guidance is interpreted and applied.     

Osteonecrosis of the jaw and the role of macrophages

Nitrogen-containing bisphosphonates have been associated with the development of osteonecrosis of the jaws (ONJ), but the lack of reliable epidemiological data and appropriate animal models has restricted our understanding of ONJ pathophysiology and limited its management. The best available information is from histopathologic findings, which implicate bone necrosis and infection, although it is not clear which is primary. However, there are data suggesting that macrophages could well be the central factor in allowing the infection to develop first, followed by local necrosis, which could also account for the development of ONJ in patients treated with denosumab, a human monoclonal antibody to the receptor activator of nuclear factor-κB ligand. This review examines the evidence that macrophages could play a prominent role in development of ONJ and the proposal that it may be more appropriate to view ONJ as a drug and not only a bisphosphonate-related complication.