下调CD133表达对胆囊癌细胞侵袭性的影响

目的:研究CD133表达与胆囊癌(gallbladder carcinoma,GBC)细胞迁移、侵袭能力的关系。方法 :用不同慢病毒转染GBC细胞,形成CD133下调的实验组和对照组,采用免疫荧光和Western印迹检测CD133表达。用划痕实验检测细胞迁移能力。用transwell实验检测细胞侵袭能力。结果:实验组细胞经慢病毒p LL3.7-sh CD133-Neo R转染后,CD133表达低于慢病毒p LL3.7-sh Lac Z-Neo R转染的对照组细胞(P<0.0.05)。结论:CD133表达与GBC细胞迁移侵袭能力相关,下调CD133表达可使GBC细胞侵袭性下降。     

CD13+CD133+和CD13-CD133-肝癌细胞的生物学差异及临床意义

目的 比较HuH7细胞系CD13+CD133+和CD13-CD133-肝细胞癌(HCC)细胞的生物学差异,并探讨其临床意义.方法 通过对CD13+CD133+ HCC细胞增殖情况、细胞周期、培养10d分化情况、裸鼠体内成瘤能力,以及对5-FU和吡柔比星等化疗药物的敏感性等生物学特征的研究,分析CD13+CD133+HCC细胞亚群的临床意义.结果 CD13+CD133+HCC细胞增殖速度明显快于CD13-CD133-HCC细胞,78.45％的CD13+CD133+HCC细胞处于G0/G1期,2.19％处于G2/M期,19.36％处于S期;62.18％的CD13-CD133-HCC细胞处于G0/G1期,11.88％处于G2/M期,25.95％处于S期.在裸鼠体内,1×103个CD13+CD133+HCC细胞即可成瘤,而1× 105个CD13-CD133-HCC才可成瘤.CD13+CD133+HCC细胞对5-FU和吡柔比星具有抵抗特性,而其他3种亚群较易被杀灭.FACS分选的CD13+CD133+HCC和CD13-CD133-HCC细胞经过10d培养后,CD13+CD133+HCC亚群的CD13、CD133标志表达与HuH7细胞系相近,而CD13-CD133-HCC细胞不具有此能力.结论 CD13+CD133+HCC细胞具有肿瘤干细胞的特征,可能与临床肝癌复发和转移有关,是临床治疗过程中尤其需要杀灭的细胞.     

Relative preservation of peripheral lung function in smoking-related pulmonary emphysema: assessment with 99mTc-MAA perfusion and dynamic 133Xe SPET

In this study the cross-sectional functional differences between the central and peripheral lung in smokers with pulmonary emphysema were evaluated by lung perfusion and dynamic xenon-133 single-photon emission tomography (SPET). The subjects were 81 patients with a long-term smoking history and relatively advanced emphysema, 17 non-smoker patients with non-obstructive lung diseases and six healthy non-smokers. Regional lung functional difference between the peripheral and central lung was assessed in the upper, middle and lower lung zones by technetium-99m macroaggregated albumin SPET and dynamic ¹³³Xe SPET. The distribution of emphysematous changes was assessed by density-mask computed tomography (CT) images which depicted abnormally low attenuation areas (LAAs) of less than –960 Hounsfield units. Two hundred and eighty-eight (59.2%) lung zones of 63 (77.7%) patients with pulmonary emphysema showed relative preservation of lung function in the peripheral lung, with a curvilinear band of normal perfusion (a stripe sign) and a significantly faster ¹³³Xe half-clearance time (T _1/2) than in central lung (P<0.0001). Of these lung zones, 256 (88.8%) showed central-dominant LAA distributions on density-mask CT images, but the remaining 32 zones did not show any regional preference in LAA distribution. Conversely, 117 (24.0%) lung zones of 19 (23.4%) patients showed periphery-dominant perfusion defects and LAA distributions, with significantly prolonged T _1/2 in the peripheral lung area (P<0.0001). The remaining 81 lung zones of the patients with pulmonary emphysema and all the lung zones of the healthy subjects and patients with non-obstructive lung diseases did not show a stripe sign, and no differences were observed in T _1/2 values and LAA distributions between the central and peripheral lung. Relative preservation of peripheral lung function seems to be a characteristic feature in smoking-related pulmonary emphysema, and may indicate a lower susceptibility of peripheral parenchyma to the development of this disease. Received 8 January and in revised form 13 March 2000     

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.     

siRNA沉默MUC5AC基因对胆管癌HCCC-9810细胞生长的影响

目的探讨特异性MUC5AC-siRNA沉默MUC5AC基因后对人肝内胆管癌细胞株HCCC-9810生长能力的影响。方法设计合成三对特异性siRNA,构建了三个稳定表达质粒,pRNAT-U6.1/Neo-MUC5AC-siRNA1/2/3,应用脂质体转染技术分别将三个稳定表达的质粒和对照质粒（空质粒对照）转染HCCC-9810,RT-PCR检测MUC5AC基因mRNA水平;SABC免疫组化染色技术检测MUC5AC粘蛋白的表达;MTT检测细胞生长增殖情况。结果基因测序表明成功构建质粒;转染后RT-PCR结果表明在mRNA水平,三个质粒都可抑制MUC5AC基因的表达,并可使MUC5AC的表达下降,MTT结果对细胞的生长有一定的抑制作用。结论构建pRNAT-U6.1/Neo-MUC5AC-siRNA1/2/3质粒明显抑制了MUC5AC基因在mRNA水平及蛋白的表达,并可抑制肿瘤细胞的生长增殖,为探讨MUC5AC在MUC5AC相关肿瘤的基因治疗方面奠定了一定的理论基础。     

Absolute activity of Ba by liquid scintillation coincidence counting using the 4π(e,X)-γ extrapolation technique

The Technical Committee for Ionizing Radiation (TCRI) of the Asia Pacific Metrology Programme (APMP) recently organized a regional key comparison of activity measurements of the radionuclide ¹³³Ba. This paper reports on absolute measurements made at the National Metrology Institute of South Africa (NMISA) by the coincidence extrapolation technique, with liquid scintillation counting (LSC) comprising the 4π channel. A detection efficiency analysis was undertaken to predict the maximum efficiency likely to be achieved and to confirm that the method does indeed provide the source disintegration rate for ¹³³Ba. Various experimental and data analysis difficulties to be aware of are discussed in the paper.     

非小细胞肺癌淋巴结转移和预后与CD133和CD147表达的关系

目的观察肿瘤干细胞标志物CD133和CD147在非小细胞肺癌（NSCLC）组织中的表达及其临床意义。方法用免疫组织化学方法检测77例NSCLC组织中CD133与CD147的表达，分析其与患者肿瘤大小、组织学类型、组织分化程度、淋巴结转移和预后的关系。结果77例NSCLC组织中CD133与CD147的阳性表达率分别为51．9％（40／77）和67．5％（52／77）。CD133和CD147的表达与淋巴结转移呈正相关（r=0．246，P〈0．05；r=0．295，P〈0．05）；CD133和CD147的表达均与患者术后生存时间呈负相关（P〈0．05）。CD133和CD147的表达与肿瘤大小、组织学类型和组织分化程度均无明显相关（P〉0．05）。CD133与CD147的表达之间呈正相关（r=0．315，P〈0．05）。结论肿瘤干细胞标志物CD133和CD147的表达与NSCLC的淋巴转移和患者预后密切相关，它们的高表达提示预后不良。     

肺腺癌A549细胞CD133~+CD44~+表型在裸鼠体内成瘤能力的研究

目的 探讨肿瘤标志物CD133和CD44的表达与人肺腺癌A549细胞株在裸鼠体内形成肿瘤能力的关系.方法 使用单细胞克隆的方法 ,筛选出能够持续增殖分裂的肿瘤细胞,应用免疫荧光方法 和流式细胞仪检测CD133和CD44在具有增殖分裂能力的A549细胞和普通A549肿瘤细胞的表达情况;按表达结果 对A549细胞进行分组,应用单克隆方法 对各组细胞进行培养,并按不间密度注射于裸鼠皮下,观察各个组别的成瘤能力.结果 (1)在倒置显微镜下观察肿瘤细胞生长情况,发现多数细胞死亡,1周后仅有4.11%的细胞能够增殖分裂形成克隆.(2)具有增殖分裂能力肿瘤细胞的免疫荧光结果 :CD133阳性率为19.0%,CD44阳性率为57.0%;普通A549肿瘤细胞免疫荧光结果 :CD133阳性率为2.5%,CD44阳性率为34.0%;具有分裂增殖能力的肿瘤细胞CD133和CD44阳性表达率显著高于普通A549肿瘤细胞(P<0.01).(3)裸鼠体内成瘤能力实验提示:CD133~+CD44~+(A组)细胞成瘤能力显著高于CD133~-CD44~-(B组)、CD133~-CD44~+(C组)和CD133~+CD44~-(D组)细胞(P<0.05);裸鼠瘤块病理检查证实均为腺癌,各脏器检查未发现转移:在接种CD133~+CD44~+(A组)细胞的瘤块组织中发现除了有CD133~+CD44~+细胞外,还有CD133~-CD44~-细胞.结论 肺腺癌A549细胞株中CD133~+CD44~+细胞在裸鼠体内的成瘤能力显著高于其他细胞.