Tumor stroma and differentiated cancer cells can be originated directly from polyploid giant cancer cells induced by paclitaxel

Paclitaxel is widely used to treat cancer patients through the blocking of mitosis and result in formation of polyploidy giant cancer cells (PGCCs), which are generally believed to be nondividing cells or in mitotic catastrophe. Here, we showed that PGCCs following the treatment of paclitaxel of MCF‐7 breast cancer cell line have capability to generate regular‐sized progeny cells through budding. The PGCCs not only grew into well‐differentiated cancer cells that formed cancer organotypic structures in vitro but also trans‐differentiated into multiple tumor stromal cells including myoepithelial, endothelial and erythroid cells. PGCCs formed glandular and vessel‐like cancer organotypic structures that expressed normal stem cell markers. These progeny cells generated from PGCCs showed decreased ability of proliferation, invasion and tumor growth and became more resistant to paclitaxel than parental MCF‐7 cells. These results demonstrated that paclitaxel‐induced PGCCs have properties of cancer stem cells that can generate both epithelial cancer cells and multilineage of stromal cells. PGCCs are not only the morphogenic determinant to tumor histogenesis and but also contribute to paclitaxel resistance.     

多烯紫杉醇诱导的卵巢癌SK-OV-3多倍体肿瘤巨细胞增殖、迁移和凋亡的特性研究

目的:探讨多烯紫杉醇(Docetaxel,Doc)诱导的卵巢癌SK-OV-3多倍体肿瘤巨细胞增殖、迁移和凋亡的特性.方法:0.8μmol/L Doc处理SK-OV-3细胞16 h,更换为完全培养液继续培养细胞至第3天和第5天.免疫荧光染色法观察细胞形态和增殖,流式细胞术检测细胞倍性、周期和凋亡,划痕实验检测细胞迁移能力...     

Number of glioma polyploid giant cancer cells (PGCCs) associated with vasculogenic mimicry formation and tumor grade in human glioma

Background

Polyploid giant cancer cells (PGCCs) contribute to solid tumor heterogeneity. This study investigated the relationships among PGCCs numbers, vasculogenic mimicry (VM) formation, and tumor grades in glioma.

Methods

A total of 76 paraffin-embedded glioma tissue samples, including 28 cases of low grade and 48 cases of high grade gliomas, were performed with H&E and immunohistochemical staining for Ki-67 and hemoglobin. The size of PGCCs nuclei was measured by a micrometer using H&E section and defined as at least three times larger than the nuclei of regular diploid cancer cells. The number of PGCCs and different blood supply patterns were compared in different grade gliomas. Microcirculation patterns in tumors were assessed using CD31 immunohistochemical and PAS histochemical double staining. Human glioma cancer cell line C6 was injected into the chicken embryonating eggs to form xenografts, which was used to observe the PGCCs and microcirculation patterns.

Results

In human glioma, the number of PGCCs increased with the grade of tumors (χ² = 4.781, P = 0.015). There were three kinds of microcirculation pattern in human glioma including VM, mosaic vessel (MV) and endothelium dependent vessel. PGCCs were able to generate erythrocytes via budding to form VM. The walls of VM were positive (or negative) for PAS staining and negative for CD31 staining. There were more VM and MVs in high grade gliomas than those in low grade gliomas. The differences have statistical significances for VM (t = 3.745, P = 0.000) and MVs (t = 4.789, P = 0.000). PGCCs, VM and MVs can also be observed in C6 chicken embryonating eggs xenografts.

Conclusions

The data demonstrated presence of PGCCs, VM and MVs in glioma and PGCCs generating erythrocytes contribute the formation of VM and MVs.     

Osteoclastoma-like giant cell tumor of the lung

The main components of an unusual form of lung tumor were osteoclast-like multinucleated giant cells and mononuclear stromal cells. Besides, scattered islands of moderately differentiated squamous cells also appeared. Both the mononuclear and the osteoclast-like giant cells reacted with antibodies against CD68 and vimentin, but did not react with antibodies against cytokeratin, EMA and CEA, or lysozyme and α-1-antitrypsin. The p53 and PCNA antigens were positive only in mononuclear cells and not the osteoclast-like giant cells, suggesting that mononuclear cells represent proliferating elements with histiocytic differentiation while osteoclast-like giant cells are stromal, presumably reactive components of the tumor.     

Oxidized high mobility group B‐1 enhances metastability of colorectal cancer via modification of mesenchymal stem/stromal cells

High mobility group box‐1 (HMGB1) is known to be a chemotactic factor for mesenchymal stem/stromal cells (MSCs), but the effect of post‐translational modification on its function is not clear. In this study, we hypothesized that differences in the oxidation state of HMGB1 would lead to differences in the function of MSCs in cancer. In human colorectal cancer, MSCs infiltrating into the stroma were correlated with liver metastasis and serum HMGB1. In animal models, oxidized HMGB1 mobilized three‐fold fewer MSCs to subcutaneous tumors compared with reduced HMGB1. Reduced HMGB1 inhibited the proliferation of mouse bone marrow MSCs (BM‐MSCs) and induced differentiation into osteoblasts and vascular pericytes, whereas oxidized HMGB1 promoted proliferation and increased stemness, and no differentiation was observed. When BM‐MSCs pretreated with oxidized HMGB1 were co‐cultured with syngeneic cancer cells, cell proliferation and stemness of cancer cells were increased, and tumorigenesis and drug resistance were promoted. In contrast, co‐culture with reduced HMGB1‐pretreated BM‐MSCs did not enhance stemness. In an animal orthotopic transplantation colorectal cancer model, oxidized HMGB1, but not reduced HMGB1, promoted liver metastasis with intratumoral MSC chemotaxis. Therefore, oxidized HMGB1 reprograms MSCs and promotes cancer malignancy. The oxidized HMGB1–MSC axis may be an important target for cancer therapy.     

口腔鳞癌肿瘤巨细胞与临床病理因素的相关性研究*

目的:探究肿瘤巨细胞（gian tcancer cell ,GCC ）在口腔鳞癌（oral squamous cell carcinoma ,OSCC）组织中的存在情况及其与OSCC临床病理因素的关系。方法:回顾性分析2005年3 月至2014年12月天津医科大学肿瘤医院76例OSCC患者的石蜡切片及临床病理资料。H&E染色及Ki-67免疫组织化学染色下观察GCC 与普通肿瘤细胞的形态差异,计数GCC 细胞,分析其与OSCC临床生物学行为及预后的关系。结果:H&E染色下观察到GCC 存在于OSCC中,且Ki-67在GCC 中均呈阳性表达。GCC 细胞数与临床分期（P = 0.02）、组织学分级（P < 0.01）、化疗（P = 0.01）及局部复发（P = 0.02）均显著相关。GCC 细胞数在不同性别和年龄的患者中差异不大,且与淋巴结转移、远处转移无关（P > 0.05）。 GCC 数目> 3 个/HP 的OSCC患者预后较差（P < 0.05）。 结论:GCC 可能在OSCC发展过程及耐药中起重要作用,可能是影响OSCC预后的重要因素。      

Circulating tumor cell levels are elevated in colorectal cancer patients with high tumor burden in the liver

Background: Metastatic spread is the most common cause of cancer-related death in colorectal cancer (CRC) patients, with the liver being the mostly affected organ. Circulating tumor cells (CTCs) are a prognostic marker in stage IV CRC. We hypothesized that tumor burden in the liver correlates with CTC quantity. Methods: Blood (7.5 ml) was prospectively collected from 24 patients with novel stage IV CRC diagnosis. Baseline EpCAM+ CTCs were analyzed with the FDA-approved CellSearch® system. Clinicopathological data were collected, and hepatic tumor burden was determined by radiographic liver volumetry with contrast-enhanced CT scans. CRC primary tumors were immunohistochemically stained for EpCAM expression with BerEP4 monoclonal antibody. Statistical analyses were performed using 2-sample T-test, non-parametric Wilcoxon Rank-Sum test, and Fisher''s exact test. Results: CTCs were detected n 17 (71%) of 24 patients. The overall mean CTC number as determined by EpCAM-based CellSearch® detection was 6.3 (SEM 2.9). High baseline CTC numbers (≥3) correlated significantly with a high tumor/liver ratio (≥30%), and with high serum CEA levels, as determined by two-sample T-test on log-transformed data and by Fisher''s Exact test on categorical data analysis (P < 0.05). The CRC primary tumors were consistently expressing EpCAM by immunostaining. Conclusions: High tumor burden in the liver and high baseline serum CEA levels are associated with high number of baseline CTCs in stage IV CRC patients. Future studies should further investigate the biological role and expression patterns of single CTCs in cancer patients to further improve personalized treatment strategies.     

非小细胞肺癌循环肿瘤细胞检测进展

肺癌是世界范围内发病率最高的恶性肿瘤, 约有18%的癌症相关死亡与肺癌有关。循环肿瘤细胞(CTCs)的出现可看做是肿瘤细胞播散入血的直接体现, 而这正是导致非小细胞肺癌患者预后较差的最重要因素之一。检测外周血中循环肿瘤细胞有望成为诊断肺癌的辅助方法, 使肺癌患者得到最佳的个体化治疗。研究者们已尝试多种不同的方法从外周血中分离CTCs。虽然有关CTCs检测的研究众多, 但不同检测方法的敏感度、特异性及重复性限制了其临床意义, 本文就现有的各项技术进行综述。      

Increased HMGB1 and cleaved caspase-3 stimulate the proliferation of tumor cells and are correlated with the poor prognosis in colorectal cancer

Background

Dying tumor cells after irradiation could promote the proliferation of living tumor cells might cause tumor relapse and treatment failure. Our previous study showed that activated caspase-3 after irradiation probably participates in tumor repopulation. In this study, we investigated whether high mobility group box 1(HMGB1) is also involved in tumor repopulation.

Methods

Colorectal tumor cells were irradiated. The cleaved caspase-3 (CC3) in irradiated tumor cells and HMGB1 in the supernatant of irradiated tumor cells were detected by Western blot. A large number of irradiated colorectal tumor cells (feeder cells) were then co-cultured with a small number of luciferase-labeled living colorectal tumor cells (reporter cells) and proliferation of reporter cells was measured by bioluminescence imaging. The CC3 and HMGB1 protein expression in colorectal tumor and peritumoral tissues were detected by immunohistochemistry and their correlation with prognosis were analyzed.

Results

The irradiated colorectal tumor cells underwent apoptosis and necrosis and produced CC3 in tumor cells and HMGB1 in the supernatant of cultured cells. The increased expression of secretory HMGB1 correlated with CC3 level and proliferating cell nuclear antigen (PCNA) after irradiation in vitro. The irradiated dying cells remarkably stimulated living tumor cell proliferation. Interestedly, immunohistochemistry staining showed that positive HMGB1, CC3, and Ki67 expression were significantly higher in colorectal tumor tissues than in peritumoral tissues (p <0.01). The Kaplan-Meier survival analysis revealed that high HMGB1, CC3, and Ki67 levels were significantly associated with poor prognosis (p <0.05, p <0.01). Multivariate analysis using Cox proportional hazards model showed that TNM staging and HMGB1 were independent prognostic factors in patients with colorectal cancer (CRC) (p <0.01, p <0.001).

Conclusion

Both apoptotic and necrotic cells could stimulate proliferation of living tumor cells, and the increased expression of CC3 and HMGB1 in tumor cells could be new markers for poor prognosis in colorectal cancer patients.

Electronic supplementary material

The online version of this article (doi:10.1186/s13046-015-0166-1) contains supplementary material, which is available to authorized users.     

Expected clinical applications of circulating tumor cells in breast cancer

Tumor cell invasion and intravascular filtration lead to the presence of circulating tumor cells (CTCs) in peripheral blood. CTCs have, thus, been counted in patients with cancer to analyze metastatic mechanisms or in the hope of developing clinical applications for diagnosis and therapy; various CTC-related studies have been performed. However, the clinical significance of CTCs remains to be established because of the extremely small number of CTCs in peripheral blood as compared with the number of blood cells. Technical problems (e.g. reproducibility and reliability) in the detection of CTCs also remain to be solved. The use of flow cytometric analysis, which can be performed with tumor-cell markers such as anti-epithelial cell adhesion molecule antibodies and anti-cytokeratin antibodies and non-tumor-cell markers such as anti-CD45 antibodies has enhanced specificity for the detection of tumor cells. The CellSearch System^® can detect 1 CTC in 7.5 mL of peripheral blood, with high reproducibility. Its detection rate and accuracy for CTCs have been confirmed. In the United States, clinical trials have used this system to detect CTCs in patients with metastatic breast cancer, metastatic colorectal cancer, and metastatic prostate cancer, and CTCs have been confirmed to be a useful prognostic factor. This system was also suggested to be useful for monitoring treatment response in patients with metastatic breast cancer and was approved by the United States Food and Drug Administration in 2004. Measuring CTC counts can facilitate the early prediction of treatment response and thereby avoid unnecessary therapy. CTCs may also be a useful biomarker for molecular targeted agents, enabling the identification of patients most likely to respond to a given treatment and facilitating treatment selection. However, the widespread use of CTC monitoring as a routine examination requires a further improvement in measurement sensitivity, the establishment of criteria for quantitative and qualitative evaluations, and additional clear-cut evidence supporting the clinical significance of CTCs. We expect that CTCs will be established to be a new diagnostic and therapeutic index for breast cancer.     

TGF‐β induced PAR‐1 expression promotes tumor progression and osteoclast differentiation in giant cell tumor of bone

Although protease activated receptor‐1 (PAR‐1) has been confirmed as an oncogene in many cancers, the role of PAR‐1 in giant cell tumor (GCT) of bone has been rarely reported. The mechanism of PAR‐1 in tumor‐induced osteoclastogenesis still remains unclear. In the present study, we detected that PAR‐1 was significantly upregulated in GCT of bone compared to normal tissues, while TGF‐β was also overexpressed in GCT tissues and could promote the expression of PAR‐1 in a dose and time dependent manner. Using the luciferase reporter assay, we found that two downstreams of TGF‐β, Smad3 and Smad4, could activate the promoter of PAR‐1, which might explain the mechanism of TGF‐β induced PAR‐1 expression. Meanwhile, PAR‐1 was also overexpressed in microvesicles from stromal cells of GCT (GCTSCs), and might be transported from GCTSCs to monocytes through microvesicles. In addition, knockout of PAR‐1 by TALENs in GCTSCs inhibited tumor growth, angiogenesis and osteoclastogenesis in GCT in vitro. Using the chick CAM models, we further showed that inhibition of PAR‐1 suppressed tumor growth and giant cell formation in vivo. Using microarray assay, we detected a number of genes involved in osteoclastogenesis as the possible downstreams of PAR‐1, which may partly explain the mechanism of PAR‐1 in GCT. In brief, for the first time, these results reveal an upstream regulatory role of TGF‐β in PAR‐1 expression, and PAR‐1 expression promotes tumor growth, angiogenesis and osteoclast differentiation in GCT of bone. Hence, PAR‐1 represents a novel potential therapeutic target for GCT of bone.     

双膦酸盐对骨巨细胞瘤细胞超微结构的影响及细胞组织学变化

目的:探讨双膦酸盐对骨巨细胞瘤细胞超微结构的影响及细胞组织学变化。方法:取我院收治的7例双膦酸盐(唑来膦酸)治疗前后的患者肿瘤组织切片进行了透视电镜观察和TUNEL凋亡染色及图像分析。结果:双膦酸盐(唑来膦酸)治疗后的肿瘤组织切片中透视电镜下多核巨细胞和基质细胞出现了明显的凋亡表现,细胞质变化的特点是大量扩张紊乱的粗面内质网及分散于细胞质中包含于囊泡中央的高电子密度核。线粒体水肿或空泡化。巨细胞核变化的特点是形成致密的染色质材料分散于细胞核中,核膜增厚或分离,部分核碎裂和形成凋亡小体;双膦酸盐(唑来膦酸)治疗后的肿瘤组织切片中TUNEL染色强烈阳性,基质细胞凋亡率从治疗前的1.31%至治疗后的33.42%,差异有显著性意义(P=0.018),多核巨细胞凋亡率从治疗前的8.41%至治疗后的56.83%,差异有显著性意义(P=0.018)。结论:双膦酸盐可以诱导骨巨细胞瘤基质细胞和多核巨细胞的凋亡。从细胞和分子生物学水平证实双膦酸盐作为骨巨细胞瘤辅助治疗方法的可行性。     

循环肿瘤细胞在肺癌中的研究进展

循环肿瘤细胞(CTC)具有与肿瘤原发灶内一致的细胞学和基因遗传学特征。 已发现CTC与肺癌的生物学行为、治疗以及预后密切相关,有望成为肺癌的又一新标记物和治疗的新靶点。本文对CTC在肺癌中的研究进展作一综述。     

Diagnosis and treatment of giant cell tumor in the thoracic spine

A 23-year-old woman with incomplete paralysis was operated upon for a giant cell tumor in the thoracic spine. X-ray films revealed a destruction lesion in the vertebral body of the 12th thoracic vertebra. On the plain thoracic computed tomography scan, the finding was a soap-bubble appearance with a linear high density area in the mass lesion which destroyed the vertebral body. Preoperative angiography showed no apparent feeding artery to the tumor tissue; preoperative myelography showed incomplete block at the level of the 12th thoracic vertebra. A radical operation was carried out in one stage via a combined antero-posterior approach. In order to give radiotherapy immediately after operation, a vascular pedicled rib graft was made. This paper discusses the role of thoracic computed tomography scan in the diagnosis of giant cell tumor and the surgical techniques used in treatment.