The role of CD133 in the identification and characterisation of tumour-initiating cells in non-small-cell lung cancer

Objective: Emerging evidence suggests that specific sub-populations of cancer cells with stem cell characteristics within the bulk of tumours are implicated in the pathogenesis of heterogeneous malignant tumours. The cells that drive tumour growth have been denoted cancer-initiating cells or cancer stem cells (hereafter CSCs). CSCs have been isolated initially from leukaemias and subsequently from several solid tumours including brain, breast, prostate, colon and lung cancer. This study aimed at isolating and characterising the population of tumour-initiating cells in non-small-cell lung cancer (NSCLC). Methods: Specimens of NSCLC obtained from 89 patients undergoing tumour resection at the Cancer National Institute of Naples were analysed. Three methods to isolate the tumour-initiating cells were used: (1) flow cytometry analysis for identification of positive cells for surface markers such as CD24, CD29, CD31, CD34, CD44, CD133 and CD326; (2) Hoechst 33342 dye exclusion test for the identification of a side-population characteristic for the presence of stem cells; (3) non-adherent culture condition able to form spheres with stem cell-like characteristics. Definition of the tumourigenic potential of the cells through soft agar assay and injection into NOD/SCID mice were used to functionally define (in vitro and in vivo) putative CSCs isolated from NSCLC samples. Results: Upon flow cytometry analysis of NSCLC samples, CD133-positive cells were found in 72% of 89 fresh specimens analysed and, on average, represented 6% of the total cells. Moreover, the number of CD133-positive cells increased markedly when the cells, isolated from NSCLC specimens, were grown as spheres in non-adherent culture conditions. Cells from NSCLC, grown as spheres, when assayed in soft agar, give rise to a 3.8-fold larger number of colonies in culture and are more tumourigenic in non-obese diabetic (NOD)/severe combined immunodeficiency (SCID) mice compared with the corresponding adherent cells. Conclusions: We have isolated and characterised a population of CD133-positive cells from NSCLC that is able to give rise to spheres and can act as tumour-initiating cells.     

Preliminary spherical agglomerates of water soluble drug using natural polymer and cross-linking technique

The objective of this study was to use the polymer, carrageenan (Gelcarin, GP 812) in an attempt to control the drug release from spheres prepared by cross-linking technique. The variables studied were the drug levels; polymer levels and the cross-linking agent. The aqueous dispersion of the drug and polymer was dropped by a disposable syringe into 3% w/v aqueous solution of cross-linking agent, and the droplets instantaneously formed spheres. Spherical agglomerates containing 77.6% w/w drug could be prepared and the physical properties of the spheres was dependent on the drug and polymer levels and also on the cross -inking material used. The drug release from spheres containing 11.9% w/w drug prepared by using 3% w/v cross-linking agent was higher than those containing 77.6% w/w drug. Carrageenan polymer can exert control over the rate and amount of drug released from spheres prepared by cross-linking technique.     

Evaluation of Bioadhesive Glipizide Spheres and Compacts from Spheres Prepared by Extruder/Marumerizer Technique

The objective of this study was to attempt to deliver glipizide from spheres and compacts containing the natural polymer Carrageenan (Gelcarin, GP 812) and prepared by extruder/marumerizer technique. A second objective was to evaluate the mucoadhesive strength of the bioadhesive spheres onto the mucus membrane of rabbit. The effects of polymer, drug level, and type of spheronizing material were evaluated. All sphere formulations were compacted into tablets using a rotary Manesty B-3B machine equipped with 12/32 flat face tooling. Results show drug release from spheres and compacts decreased as the level of Carrageenan was increased. However, as the level of drug was increased drug release also increased. Spheres containing Avicel PH-101 gave higher drug release than spheres of the same composition but prepared with Avicel RC-581. In general, the drug release from tablets was higher than its corresponding spheres and drug release from spheres and tablets containing Carrageenan was higher than control spheres and tablets of the same composition but without Carrageenan. Tablet formulations compacted from spheres containing Avicel RC-581 gave higher release rate constants than tablet formulations of the same composition but prepared with Avicel PH-101. The bioadhesion study showed that mucoadhesion strength between spheres and mucus membrane of the rabbit depends on the levels of polymer, drug, and type of spheronizing material. Developed bioadhesive spheres and tablets increase the solubility of glipizide which may increase its bioavailability and also increased the adherence of the bioadhesive systems to the mucous membrane so that once daily dose can be administered.     

胃癌干细胞的分离鉴定以及干细胞特性研究

背景:肿瘤干细胞(CSCs)已成为当前肿瘤研究的热点之一,开展相关研究的首要问题是CSCs的分离和鉴定,悬浮培养法是分离CSCs的重要方法。目的:分离、鉴定胃癌干细胞(GCSCs)并评价其干细胞特性。方法:人胃癌细胞株MKN45、MGC803、SGC7901以含表皮生长因子(EGF)、碱性成纤维细胞生长因子(bFGF)的无血清培养基悬浮培养。对培养得到的第三代肿瘤球,以集落形成实验检测其克隆形成能力,免疫荧光法检测GCSCs标记物,细胞划痕实验和Transwell小室细胞侵袭实验检测其迁移、侵袭能力,CCK-8实验检测其对顺铂的耐药性。结果:培养得到的肿瘤球高表达干细胞标记物CD44、CD54,低表达胃壁细胞标记物H+/K+-ATPase,克隆形成能力、划痕愈合速度和Transwell小室穿膜细胞数明显大于或多于普通胃癌细胞,差异有统计学意义(P0.05)。顺铂对肿瘤球的50%抑制浓度明显高于普通胃癌细胞。结论:应用无血清培养基悬浮培养法能成功获得GCSCs富集的肿瘤球;肿瘤球具有较强的自我更新、增殖、迁移、侵袭能力和多向分化潜能,对化疗药物更易产生耐药性。     

膀胱癌T24中肿瘤干细胞样细胞的分离与鉴定

目的观察使用无血清培养基悬浮培养能否从膀胱癌T24中分离出微球体,并证实其是否有肿瘤干细胞的特点。方法使用无血清培养基DMEM/F12添加生长因子EGF、bFGF、胰岛素、B27对膀胱癌T24进行悬浮培养,并从长期增殖能力、克隆形成能力、自我更新能力、对放化疗的抵抗能力、迁移和侵袭能力等方面证实这些微球体是否有肿瘤干细胞的特点。结果从培养的第4天开始长出微球体,微球体细胞比普通贴壁T24细胞有更强的增殖能力和克隆形成能力,对放化疗的抵抗能力更强,有更强的迁移和侵袭能力,微球体有自我更新能力,具有肿瘤干细胞的特点。结论使用无血清悬浮培养可以从T24中分离出微球体,而且这些微球体细胞有肿瘤干细胞的特点。     

Treatment options for unresectable HCC with a focus on SIRT with Yttrium‐90 resin microspheres

Hepatocellular carcinoma (HCC), the predominant form of primary liver cancer, is the second leading cause of cancer‐related deaths across the globe. Only a small percentage of HCC patients (~20%‐30%) are diagnosed at an early stage when first‐line treatment options may be effective. The majority of HCC patients (>70%) are diagnosed with unresectable disease and given a poor overall prognosis. Current treatment guidelines recommend locoregional therapy with transarterial chemoembolisation (TACE) and systemic therapy with sorafenib as first‐line treatment for patients with intermediate and advanced stage HCC. However, multiple factors including contraindications, technical considerations and treatment‐related toxicities pose significant challenges in achieving favourable treatment outcomes, underscoring the need for a paradigm shift in managing these patients. In 2002, yttrium‐90 (Y‐90) resin microspheres was approved by the U.S. Food and Drug Administration (FDA) for the treatment of unresectable metastatic colorectal cancer to the liver with adjuvant floxuridine chemotherapy. However, thousands of patients with unresectable HCC have also been treated with resin Y‐90. For over two decades, several small‐scale prospective trials and retrospective studies have investigated and reported on the efficacy of locoregional selective internal radiation therapy (SIRT) with Y‐90 microspheres in treating unresectable HCC. Although it is currently a treatment option for intermediate‐stage HCC patients, mainstream clinical application of resin Y‐90 has been largely limited because of the lack of sufficient clinical data from a randomised controlled trial. This could change with the imminent announcement of results from the phase 3 Sorafenib vs Radioembolization in Advanced Hepatocellular carcinoma (SARAH) trial. To provide the foundation and context for interpreting results from the SARAH trial, this article provides an overview of treatment modalities and current challenges in managing unresectable HCC. There is also a review of key prospective and retrospective studies evaluating the use of Y‐90 SIRT, specifically Y‐90 resin microspheres in unresectable HCC, which led to the development of the SARAH trial. Methods: To identify relevant publications, the PubMed database was queried using one or more of the following search terms alone or in combination with Boolean operators: epidemiology, hepatocellular, hepatocellular cancer, hepatocellular carcinoma, unresectable, radioembolisation, selective internal radiation therapy, SIR‐Spheres, yttrium 90, TACE, and sorafenib. The results were sorted or filtered by “Author”, “Publication dates” or “Article types” to identify articles relevant to each section of the review. To ensure that information on ongoing clinical trials involving Y‐90 resin was included, we conducted a search on “ClinicalTrials.gov”, by combining the search terms “HCC” OR “hepatocellular carcinoma” with “Y 90” OR “yttrium 90” OR “radioembo”, and screened for studies that involved treatment with Y‐90 resin microspheres.     

吸毒者的心理控制源及相关分析

目的:探讨吸毒者的心理控制源的特点,采取针对性的干预措施。方法:采用控制圈量表(SOC)对86例吸毒者在脱毒后一个月进行问卷调查,和66名打工者进行比较分析,并且做了初吸者与复吸者的比较,采用SPSS10.0软件包进行统计处理。结果:吸毒者个人实力量表评分显著低于对照组,人际控制和社会政治控制量表评分显著高于对照组,初吸者与复吸者比较,初吸者个人实力量表评分显著高于对照组,人际控制和社会政治控制量表评分无显著性差异,相关分析表明个人实力量表评分与吸毒时间及戒毒次数负相关。结论:吸毒者较少认为事情的结果与个人实力有关,自我控制能力较低,而吸毒者的心理控制源的特点又容易产生心理问题,提示对吸毒者必须采取针对性的干预措施,改变其心理控制源的特点,以降低复吸率。     

Treatment of unresectable cholangiocarcinoma using yttrium-90 microspheres: results from a pilot study

BACKGROUND.

The objective of this report was to present data from an open‐label cohort study in which patients with intrahepatic cholangiocarcinoma (ICC) underwent radioembolization with yttrium‐90 (⁹⁰Y) microspheres.

METHODS.

Twenty‐four patients with histologically proven ICC were treated. The planned target dose was 120 Gray. Patients were stratified according to Eastern Cooperation Oncology Group (ECOG) performance status, tumor morphology (infiltrative vs peripheral), tumor distribution (solitary vs multifocal), and the presence or absence of portal vein thrombosis (PVT). Before and after the procedure, the following variables were assessed: 1) biochemical and clinical toxicity, 2) imaging (computed tomography/magnetic resonance imaging) response according to World Health Organization and European Association for the Study of Liver Disease (EASL) criteria, and 3) median survival after the first treatment using Kaplan‐Meier methodology.

RESULTS.

In total, 48 ⁹⁰Y treatments were administered to hepatic segments or lobes. Fatigue and transient abdominal pain were reported in 18 patients (75%) and 10 patients (42%), respectively. One patient (4%) developed grade 3 bilirubin toxicity. One patient (4%) developed a treatment‐related gastroduodenal ulcer. On imaging follow‐up of 22 patients, tumors demonstrated a partial response in 6 patients (27%), stable disease in 15 patients (68%), and progressive disease in 1 patient (5%). By using EASL guidelines, 17 patients (77%) showed >50% tumor necrosis on imaging follow‐up. Two patients (9%) demonstrated 100% tumor necrosis. The median overall survival for the entire cohort (n = 24) was 14.9 months. The median survival for patients with an ECOG performance status of 0, 1, and 2 was 31.8 months, 6.1 months, and 1 month, respectively (P < .0001); the median survival for patients without and with PVT was 31.8 months and 5.7 months, respectively (P = .0003); and the median survival for patients with peripheral versus periductal‐infiltrative tumors was 31.8 months and 5.7 months, respectively (P = .0005).

CONCLUSIONS.

Radioembolization with ⁹⁰Y may be a therapeutic option for the treatment of unresectable ICC. Cancer 2008. © 2008 American Cancer Society.