Bystander effect in tumor cells produced by Iodine-125 labeled human lymphocytes

Abstract

Purpose: To investigate the ability of human lymphocytes labeled with DNA-incorporated ¹²⁵I to exert an inhibitory (antiproliferative) bystander effect on co-cultured human colon adenocarcinoma LS174T cells in vitro.

Materials and methods: Human peripheral blood lymphocytes were stimulated to synthesize DNA in the presence of phytohemagglutinin (PHA) and labeled with 5-[¹²⁵I]iodo-2′-deoxyuridine. Human colon adenocarcinoma LS174T cells were co-cultured with the ¹²⁵I-labeled lymphocytes in various ratios for 5 days and the proliferation of the LS174T cells was assessed. Further, the supernatant media from these co-cultures were: (i) Transferred to LS174T cells and their proliferation measured after 5 days, (ii) used to assess the clonogenic survival of LS174T cells, and (iii) screened for factors that suppress growth.

Results: A significant reduction in the proliferation of LS174T cells was observed when co-cultured either with ¹²⁵I-labeled lymphocytes (56?±?3.5%) or the supernatant media (52.5?±?1.3%) obtained from these co-cultures. Clonogenic survival of LS174T cells grown in the supernatant media corroborated the decrease in tumor cell growth.

Conclusion: The observed reduction in the proliferation of LS174T cells in presence of ¹²⁵I-labeled lymphocytes or media obtained from such co-cultures can be attributed to an inhibitory (antiproliferative) bystander effect, probably mediated by factor(s) released from the dying ¹²⁵I-labeled lymphocytes.     

Irradiated human endothelial progenitor cells induce bystander killing in human non-small cell lung and pancreatic cancer cells

Purpose To investigate whether irradiated human endothelial progenitor cells (hEPC) could induce bystander killing in the A549 non-small cell lung cancer (NSCLC) cells and help explain the improved radiation-induced tumor cures observed in A549 tumor xenografts co-injected with hEPC.

Materials and methods We investigated whether co-injection of CBM3 hEPC with A549 NSCLC cells would alter tumor xenograft growth rate or tumor cure after a single dose of 0 or 5?Gy of X-rays. We then utilized dual chamber Transwell dishes, to test whether medium from irradiated CBM3 and CBM4 hEPC would induce bystander cell killing in A549 cells, and as an additional control, in human pancreatic cancer MIA PaCa-2 cells. The CBM3 and CBM4 hEPC were plated into the upper Transwell chamber and the A549 or MIA PaCa-2 cells were plated in the lower Transwell chamber. The top inserts with the CBM3 or CBM4 hEPC cells were subsequently removed, irradiated, and then placed back into the Transwell dish for 3?h to allow for diffusion of any potential bystander factors from the irradiated hEPC in the upper chamber through the permeable membrane to the unirradiated cancer cells in the lower chamber. After the 3?h incubation, the cancer cells were re-plated for clonogenic survival.

Results We found that co-injection of CBM3 hEPC with A549 NSCLC cells significantly increased the tumor growth rate compared to A549 cells alone, but paradoxically also increased A549 tumor cure after a single dose of 5?Gy of X-rays (p?p?p?p?Conclusions These data provide evidence that irradiated hEPC can induce strong bystander killing in A549 and MIA PaCa-2 human cancer cells and that this bystander killing is mediated by the cytokines TNF-α and TGF-β.     

Lack of evidence for low-LET radiation induced bystander response in normal human fibroblasts and colon carcinoma cells

Purpose:?To investigate radiation-induced bystander responses and to determine the role of gap junction intercellular communication and the radiation environment in propagating this response.

Materials and methods:?We used medium transfer and targeted irradiation to examine radiation-induced bystander effects in primary human fibroblast (AG01522) and human colon carcinoma (RKO36) cells. We examined the effect of variables such as gap junction intercellular communication, linear energy transfer (LET), and the role of the radiation environment in non-targeted responses. Endpoints included clonogenic survival, micronucleus formation and foci formation at histone 2AX over doses ranging from 10–100 cGy.

Results:?The results showed no evidence of a low-LET radiation-induced bystander response for the endpoints of clonogenic survival and induction of DNA damage. Nor did we see evidence of a high-LET, Fe ion radiation (1 GeV/n) induced bystander effect. However, direct comparison for 3.2 MeV α-particle exposures showed a statistically significant medium transfer bystander effect for this high-LET radiation.

Conclusions:?From our results, it is evident that there are many confounding factors influencing bystander responses as reported in the literature. Our observations reflect the inherent variability in biological systems and the difficulties in extrapolating from in?vitro models to radiation risks in humans.     

SKP2表达对受照食管癌细胞诱导辐射旁效应的影响

目的：探究SKP2表达水平对受照食管癌细胞所诱导辐射旁效应的影响。方法：Western blot法筛选出SKP2高表达和低表达的食管癌细胞系,微核检测法探讨SKP2对受照食管癌细胞所诱导辐射旁效应的影响。建立SKP2高表达和低表达的转染细胞系,Western blot法验证转染效率,微核检测法进一步验证SKP2对受照食管癌细胞所诱导辐射旁效应的影响。通过γ-H2AX聚集点检测法探究SKP2影响受照食管癌细胞所诱导辐射旁效应的作用机制。结果：微核检测结果表明,SKP2高表达的受照细胞诱导的辐射旁效应显著低于SKP2低表达的受照细胞诱导的辐射旁效应（t=8.06,P<0.01）。SKP2转染细胞系的微核检测结果进一步验证了SKP2的表达对受照食管癌细胞诱导的辐射旁效应的抑制作用,SKP2表达升高,辐射旁效应减弱（t=11.12、10.16,P<0.01）;SKP2表达降低,辐射旁效应增强（t=8.39、8.83,P<0.01）。γ-H2AX聚集点检测结果表明,受照细胞SKP2表达升高可提高旁效应细胞的DNA损伤修复能力（t=6.85、7.10,P<0.01）。反之,会抑制旁效应细胞的DNA损伤修复能力（t=7.66、8.47,P<0.01）。结论：SKP2的表达抑制受照食管癌细胞诱导的辐射旁效应,并且这一机制至少部分是通过SKP2对DNA损伤修复能力的调节来实现的。     

血清CA153、CA125及CA199检测对乳腺癌的诊断价值

目的:探讨CA153、CA125、CA199联合检测在乳腺癌诊断中的价值。方法:采用电化学发光法分别检测乳腺癌、乳腺良性疾病患者、体检健康者血清中CA153、CA125、CA199的水平。结果:乳腺癌患者血清CA153、CA125、CA199、水平均显著高于正常组和良性乳腺疾病组,差异有统计学意义(P<0.01)。CA153、CA125、CA199、单独检测乳腺癌的灵敏度分别为55%、38%、36%,三者联合检测乳腺癌的灵敏度为78.0%。三项联合检测与单独检测相比灵敏度明显(P<0.05)。结论:血清CA153、CA125及CA199单项检测对乳腺癌的诊断均有较大意义,联合检测血清CA153、CA125及CA199能提高乳腺癌的诊断率。     

125I粒子照射诱导人前列腺癌细胞凋亡的实验研究

目的探讨^125I粒子持续低剂量率照射诱导人前列腺癌细胞凋亡的作用机制。方法用DNA琼脂糖凝胶电泳和流式细胞仪分析^125I粒子持续低剂量率照射对PC3细胞的凋亡诱导作用，检测天冬酰胺特异酶切的半胱氨酸蛋白酶Caspase-3活性来观察其对PC3细胞的凋亡诱导途径，并通过间接免疫荧光技术检测其对Bcl-2表达的影响。结果DNA琼脂糖凝胶电泳可观察到清晰的“DNA梯度”，流式细胞仪检测^125I粒子持续低剂量率照射对PC3细胞具有明显的凋亡诱导作用。Caspase-3活性检测表明，Caspase-3活性无明显变化。流式细胞仪分析表明Bcl-2的表达随^125I粒子剂量的增加而下降。结论^125I粒子持续低剂量率照射可诱导PC3细胞凋亡，其诱导凋亡与Bcl-2表达有关，与Caspase-3活性无关。     

Magnetic resonance imaging for monitoring the effects of thalidomide on experimental human breast cancers

Thalidomide, which inhibits angiogenesis in certain tumor types, reduced extravasation of a macromolecular contrast medium (MMCM) in a human breast cancer model as assayed by MMCM-enhanced dynamic magnetic resonance imaging (MRI) and fluorescence microscopy in the same tumors. After a 1-week, three-dose course of thalidomide, the mean MRI-assayed endothelial transfer coefficient, K^PS, decreased significantly (p < 0.05) from 19.4 ± 9.1 to 6.3 ± 9.1 μl/min·100 cm³. Correspondingly, microscopic measurements of extravasated MMCM, expressed as fractional area of streptavidin staining, were significantly (p < 0.05) lower in thalidomide-treated tumors (18.6 ± 11.9%) than in control saline-treated tumors (50.2 ± 2.3%). On a tumor-by-tumor basis, post-treatment K^PS values correlated significantly (r ² = 0.55, p < 0.05) with microscopic measures of MMCM extravasation. However, no significant differences were observed between saline- and thalidomide-treated tumors with respect to rate of growth, vascular richness, or amount of VEGF-containing cells. Because of its sensitivity to the detection of changes in vascular leakage in tumors, this MMCM-enhanced MRI assay could prove useful for monitoring the effects of thalidomide on an individual patient basis. The significant correlation between MRI and fluorescence microscopic measures of MMCM extravasation supports the utility of the non-invasive MRI approach for assessing the action of thalidomide on tumor blood vessels.     

乳腺癌两种手术方式对女性婚姻质量的影响

 目的 比较乳腺癌保乳手术和改良根治术对女性婚姻质量的影响.方法 同期选取接受保乳手术和改良根治术的乳腺癌患者各28例,在手术后6个月,采用自编凋查问卷和Olson婚姻质量问卷(ENRICH)进行调查.结果 两种乳腺癌手术患者的婚姻质量在婚姻满意度、夫妻交流、性生活3个方面得分低于常模(P<0.01);保乳手术患者在婚姻质量的3个维度得分均高于改良根治术患者(P<0.01).结论 乳腺癌手术后患者婚姻质量下降.保乳手术患者婚姻质量优于改良根治术患者,保乳手术对于保障乳腺癌女性婚娟质量有一定作用.     

简便人大肠癌原代细胞培养方法的探讨

目的探讨简便、实用的人大肠癌原代细胞培养方法。方法在培养瓶底放置经过高浓度抗生素和两性霉素B反复处理的约1 mm3大小的人大肠癌组织块数个,加入2 ml含高浓度抗生素和两性霉素B的RPMI 1640培养液,直立培养7～10 h,轻轻放平培养24 h后再加入2 ml高浓度抗生素和两性霉素B的RPMI 1640培养液,至大量肿瘤细胞培养出来。结果肿瘤细胞在3～4 d后从组织块内爬出,贴壁大量生长,细胞融合成片,蔓延至整个瓶底。结论 利用简便易行的方法对大肠癌新鲜标本进行原代细胞培养可以得到存活的肿瘤细胞。     

黄荆子乙酸乙酯提取物对人乳癌细胞体内外实验的研究

目的研究黄荆子乙酸乙酯提取物（EVn-50）在体内外对人乳癌MDA—MB-435S细胞的抑制作用。方法软琼脂克隆形成法测定细胞生长：溴化尿嘧啶（BrdU）掺入法检测细胞核酸的合成；建立MDA—MB-435S裸鼠异种移植瘤模型，观察经EVn-50处理后的肿瘤抑制率。结果在体外，随EVn-50浓度增高，细胞集落形成率明显下降（P〈0．01）；EVn-50对MDA—MB-435S细胞核酸合成具有抑制作用，呈剂量依赖性，其中高剂量组的BrdU掺入抑制作用最强，IC50为16．8ug／mL。在体内，EVn-50抑制MDA—MB-435S细胞裸鼠异种移植瘤生长，20、40、80ug／mL的EVn-50对移植瘤的瘤重抑制率分别为18％、31％和63％，呈浓度依赖性。结论EVn-50在体内外对MDA—MB-435S细胞均具有生长抑制作用。     

Oestrogen-mediated regulation of somatostatin receptor expression in human breast cancer cell lines assessed with 99mTc-depreotide

Investigating three somatostatin receptor (SSTR)-positive (+) human breast cancer cell lines, Xu et al. found a time- and dose-dependent up- or down-regulation of SSTR2 mRNA expression by 17-oestradiol (E₂) or the anti-oestrogen tamoxifen, respectively, in the two oestrogen receptor-positive (ER+) cell lines but not in the oestrogen receptor-negative (ER–) cell line. This study aimed to confirm the findings of Xu et al. at the protein level by means of western blotting and saturation binding studies using ^99mTc-depreotide (NeoSpect). The ER+/SSTR+ ZR75-1 and T47D and SSTR+/ER– MDA MB231 breast cancer cell lines were exposed to 1 nM E₂ or a combination of 1 nM E₂ plus 100 nM tamoxifen or ICI 182 780 (Faslodex) for 48 h. Exposed and non-exposed controls were incubated with increasing concentrations of ^99mTc-depreotide (0.5 nM–15 nM) in the absence and the presence of 20 M of octreotide. Scatchard-Rosenthal plots were derived using commercially available software. SSTR subtypes responsible for E₂-induced changes in ^99mTc-depreotide binding were identified by means of western blotting. Mean K_d values for ^99mTc-depreotide were 13 nM, 7 nM and 4 nM for T47D, ZR75-1 and MDA MB231 cells, respectively. After stimulation with E₂, the ER+ cell line T47D demonstrated a mean increase of 81% (P<0.05) in ^99mTc-depreotide binding. Adding the partial agonist tamoxifen and full antagonist ICI 182 780 to E₂ blocked the induced increase in T47D cells, either reducing SSTR expression or restoring it to control levels. ZR75-1 cells stimulated with E₂ showed a mean decrease in ^99mTc-depreotide binding of 36% as compared to control cells; this difference, however, proved to be not statistically significant. Similarly, B_max values did not change in ZR75-1 cells exposed to E₂ in combination with an ER antagonist as compared to control cells. Finally, no influence of E₂ on ^99mTc-depreotide binding was observed in the ER– cell line MDA MB231. Both SSTR2 and SSTR5 were expressed at high levels in T47D cells and ZR75-1 cells. SSTR5 drastically increased in the absence of E₂ and was restored to the original detection level after E₂ treatment. The presented findings support an oestrogen-dependent regulation of SSTR expression in breast cancer cell lines.     

超声造影对乳腺癌患者新辅助化疗效果的评估价值

目的 研究超声造影对乳腺癌患者新辅助化疗效果的评估价值,与核磁共振灌注成像和彩色多普勒超声做比较.方法 选取2011年1月～2014年11月在我院接受新辅助化疗的乳腺癌患者60例,在化疗前后均进行超声造影、核磁共振灌注成像和彩色多普勒超声检查,将检查结果与病理诊断结果做比较,评估3种诊断方式的准确性.结果 60例乳腺癌患者经新辅助化疗后,病理诊断结果为完全缓解16例,部分缓解29例,无缓解15例;超声造影检查结果为完全缓解25例,部分缓解23例,无缓解12例;核磁共振灌注成像检查结果为完全缓解22例,部分缓解25例,无缓解13例;彩色多普勒超声检查结果为完全缓解27例,部分缓解23例,无缓解10例.三者与病理诊断结果比较,McNemar-Bowker检验结果显示,P值均＞0.05,显示三者的诊断结果与病理检查结果均有一定的相关性;Kappa检验结果显示,核磁共振灌注成像结果与病理检查结果一致性最好.结论 超声造影、核磁共振灌注成像和彩色多普勒超声对乳腺癌患者新辅助化疗效果均有一定的评估价值,核磁共振灌注成像与病理诊断结果的一致性最好,可作为首选,超声造影可作为疗效评价的辅助手段.     

Response of choline metabolites to docetaxel therapy is quantified in vivo by localized (31)P MRS of human breast cancer xenografts and in vitro by high-resolution (31)P NMR spectroscopy of cell extracts.

Choline-containing compounds (CCCs) are elevated in breast cancer, and detected in vivo by the (1)H MRS total choline (tCho) resonance (3.25 ppm) and the (31)P MRS phosphomonoester (PME) resonance (3.8 ppm). Both the tCho and PME resonances decrease early after initiation of successful therapy. The single major component of these composite resonances, phosphocholine (PCho), also responds to therapy by decreasing. The ability to resolve and quantify PCho in vivo would thus increase the sensitivity of this biomarker for early detection of therapeutic response. Herein, the in vivo resolution and quantification of PCho is reported in human mouse xenograft tumors of the human breast cancer cell lines MCF-7 and MDA-mb-231. Significant decreases in tumor PCho are observed within 2 to 4 d posttreatment with the antimicrotubule drug, docetaxel. To determine whether these decreases are a general tumor response or an intracellular metabolic response, high-resolution NMR spectroscopy was performed on extracts of cells treated with docetaxel. Significant decreases in intracellular PCho and increases in glycerophosphocholine (GPC) were observed. These decreases are coincident with other tumor and cellular responses such as tumor growth delay (TGD), cell-cycle arrest, and modes of cell death such as mitotic catastrophe, necrosis, and apoptosis, with mitotic catastrophe predominating.     

两种化疗方案对转移性乳腺癌外周血T细胞亚群 和临床预后影响的比较

 目的 探讨贝伐单抗联合曲妥珠单抗+紫杉醇(TH)方案化疗对人类表皮生长因子受体2(human epidermal growth factor receptor-2, Her-2)阳性的转移性乳腺癌患者外周血T细胞亚群和临床预后的影响。方法 选取Her-2阳性的乳腺癌患者100例,随机分为研究组和对照组;研究组采用贝伐单抗联合TH方案化疗,对照组仅采用TH方案化疗。随访终点为2年,主要观察指标为CD4⁺T细胞、CD8⁺T细胞、实体瘤疗效评价等级、无进展生存期和2年病死率。结果 两组治疗前后CD8⁺T细胞差异均无统计学意义。两组治疗前CD4⁺T细胞差异无统计学意义(P=0.422)。治疗后,研究组CD4⁺T细胞显著高于对照组(P=0.011)。研究组完全缓解、部分缓解、病情稳定和病情进展发生率分别为0.0%、36.0%、42.0%和22.0%,对照组为0.0%、18.0%、44.0%和38.0%。2年后,研究组9例死亡,病死率为18.0%,对照组20例死亡,病死率为40.0%,差异有统计学意义(P=0.015)。Wilcoxon检验显示研究组无进展生存期显著高于对照组(P=0.007)。结论 贝伐单抗联合TH方案化疗有助于改善Her-2阳性的转移性乳腺癌患者免疫功能和临床预后。     

Anticancer effects elicited by combination of Rubus extract with phthalocyanine photosensitiser on MCF-7 human breast cancer cells

BackgroundPhotodynamic therapy (PDT) is a novel approach for the treatment of cancer and other related diseases. Breast cancer remains the most common cause of cancer-related death in women. This study was carried out to investigate the photosensitizing capacity of Rubus fairholmianus root acetone extract (RFRA) in vitro.MethodsRFRA was coupled with phthalocyanine photosensitizer to enhance the therapeutic properties on MCF-7 breast cancer cells. Comparatively low dose photosensitizer (PS) and Rubus extract have been used for the conjugation as it induces cell death at low doses. The diode laser of wavelength 680 nm and 5, 10 and 15 J/cm2 fluencies have been used for PDT experiments/laser irradiation. MCF-7 cells were exposed to Rubus extract and conjugated Rubus-PS for 24 h and analysed the alterations in cell morphology, proliferation, cytotoxicity and apoptosis induction.ResultsThe PDT-treated cells displayed substantial features of apoptotic cell death by changes in morphology with a reduction in cell number, development of apoptotic bodies and cell detachment from culture plates. Cellular viability (51.25% for RFRA-PS at 15 J/cm2) and Adenosine 5′-triphosphate (ATP) proliferation of treated cells reduced significantly and the cytotoxicity increased in lactate dehydrogenase (LDH) assay. The Annexin V/PI double staining supports the caspase 3/7 activities by the increased apoptotic cells population and the increased levels of cytochrome c.ConclusionOur results show that the phototoxic properties of RFRA and photosensitizer may be through the caspase-mediated apoptosis and it can be summarised that Rubus may be a potent anticancer plant with phototoxic effects on breast cancer cells.     

釉基质蛋白和骨形成蛋白对牙周膜成纤维细胞增殖活性的影响

目的:通过检测釉基质蛋白(Enamel matrix proteins,EMPs)、骨形成蛋白-2(BMP-2)作用下对促进牙周膜成纤维细胞增殖活性的影响,探讨EMPs、BMP-2促进牙周组织再生的作用机制及其之间的相互影响.方法:酶消化法获得人牙周膜成纤维细胞(Haman periodontal ligament cells,HPDLC)用不同浓度的因子进行诱导,在3d、7d两个时间点,利用MTT法检测和分析诱导后细胞增殖活性的变化.结果:BMP-2、EMPs单独应用均可促进人PDLCs增殖,BMP-2作用强于EMPs.而联合组与100～200μg/L BMP-2、200 mg/L EMPs无明显差异,与100 mg/L EMPs差异显著.结论:BMP-2、EMPs单独或联合应用均可促进人PDLCs增殖,BMP-2作用强于EMPs.联合组中起主要作用的是BMP-2,EMPs未能增加BMP-2的作用.     

重组人血管内皮抑制素（恩度）治疗非小细胞肺癌研究进展

重组人血管内皮抑素注射液（rh-endostatin,YH16）,商品名为恩度（EndostarTM）是我国学者自主研发的一种新型重组人血管内皮抑制药物,临床前研究显示该药能抑制血管内皮细胞增殖、血管生成和肿瘤生长。Ⅰ期临床结果证明该药单用临床应用安全有效。Ⅱ、Ⅲ期临床研究表明恩度与化疗方案联合具有协同作用,能明显提高晚期非小细胞肺癌（NSCLC）的有效率及中位疾病进展时间,且安全性较好,不增加化疗的不良反应,是晚期NSCLC的一种安全、有效的治疗方案。该文综述了近年来恩度治疗NSCLC的临床研究及应用进展。     

γ射线和甲基胆蒽对诱发人胚肺细胞转化的合并作用

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