Fluorescence-Activated Cell Sorting Analysis of Heterotypic Cell-in-Cell Structures

He, Mingbo; Huang, Hongyan; Wang, Manna; Chen, Ang; Ning, Xiangkai; Yu, Kaitao; Li, Qihong; Li, Wen; Ma, Li; Chen, Zhaolie; Wang, Xiaoning; Sun, Qiang

doi:10.1038/srep09588

Cited by 16 publications

(16 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We previously reported the method to quantify homotypic CICs by immunostaining and microscopic observation [ 28 ], which had been widely used for CICs studies. And lately, we developed a high-through method to quantify heterotypic CICs based on flow cytometry analysis, which turned out to be quite efficient to enrich CICs with purity higher than 90% [ 29 ]. In this work, we described a method, named EML method based on cell types examined, to analyze CICs in tissues.…”

Section: Discussionmentioning

confidence: 99%

Detecting cell-in-cell structures in human tumor samples by E-cadherin/CD68/CD45 triple staining

Huang

Chen²,

Wang

et al. 2015

Oncotarget

Self Cite

View full text Add to dashboard Cite

Although Cell-in-cell structures (CICs) had been documented in human tumors for decades, it is unclear what types of CICs were formed largely due to low resolution of traditional way such as H&E staining. In this work, we employed immunofluorescent method to stain a panel of human tumor samples simultaneously with antibodies against E-cadherin for Epithelium, CD68 for Macrophage and CD45 for Leukocytes, which we termed as “EML method” based on the cells detected. Detail analysis revealed four types of CICs, with tumor cells or macrophage engulfing tumor cells or leukocytes respectively. Interestingly, tumor cells seem to be dominant over macrophage (93% vs 7%) as the engulfer cells in all CICs detected, whereas the overall amount of internalized tumor cells is comparable to that of internalized CD45+ leukocytes (57% vs 43%). The CICs profiles vary from tumor to tumor, which may indicate different malignant stages and/or inflammatory conditions. Given the potential impacts different types of CICs might have on tumor growth, we therefore recommend EML analysis of tumor samples to clarify the correlation of CICs subtypes with clinical prognosis in future researches.

show abstract

Section: Discussionmentioning

confidence: 99%

Detecting cell-in-cell structures in human tumor samples by E-cadherin/CD68/CD45 triple staining

Huang

Chen²,

Wang

et al. 2015

Oncotarget

Self Cite

View full text Add to dashboard Cite

show abstract

“…Although being extensively reported in human tumors for a long time, the mechanisms underlying CICs formation remained unexplored until recent years, when several research models were established 8 12 13 . Meanwhile, the research methods were also being developed for both homotypic and heterotypic CICs 18 19 , and for subtyping CICs in human tumor samples 20 . Currently, it is well accepted that cell-cell adhesion and cytoskeleton are two key players in CICs formation.…”

Section: Discussionmentioning

confidence: 99%

Impaired formation of homotypic cell-in-cell structures in human tumor cells lacking alpha-catenin expression

Wang

Ning

Chen³

et al. 2015

Sci Rep

Self Cite

View full text Add to dashboard Cite

Although cell-in-cell structures (CICs) could be detected in a wide range of human tumors, homotypic CICs formed between tumor cells occur at low rate for most of them. We recently reported that tumor cells lacking expression of E- and P-cadherin were incapable of forming homotypic CICs by entosis, and re-expression of E- or P-cadherin was sufficient to induce CICs formation in these tumor cells. In this work, we found that homotypic CICs formation was impaired in some tumor cells expressing high level of E-cadherin due to loss expression of alpha-catenin (α-catenin), a molecular linker between cadherin-mediated adherens junctions and F-actin. Expression of α-catenin in these tumor cells restored cell-cell adhesion and promoted CICs formation in a ROCK kinase-dependent way. Thus, our work identified α-catenin as another molecule in addition to E- and P-cadherin that were targeted to inactivate homotypic CICs formation in human tumor cells.

show abstract

“…The fate of inner cells in cell‐in‐cell structures had been reported to be partially dependent on the properties of the invading cells . We found emperipolesis of CD8 T cells appeared to be related with apoptosis and proliferation of BECs.…”

Section: Discussionmentioning

confidence: 99%

Emperipolesis mediated by CD8⁺ T cells correlates with biliary epithelia cell injury in primary biliary cholangitis

Zhao

et al. 2019

J Cellular Molecular Medi

View full text Add to dashboard Cite

Primary biliary cholangitis (PBC) is an autoimmune disease characterized by chronic destruction of the bile ducts. A major unanswered question regarding the pathogenesis of PBC is the precise mechanisms of small bile duct injury. Emperipolesis is one of cell‐in‐cell structures that is a potential histological hallmark associated with chronic hepatitis B. This study aimed to clarify the pathogenesis and characteristics of emperipolesis in PBC liver injury. Sixty‐six PBC patients, diagnosed by liver biopsy combined with laboratory test, were divided into early‐stage PBC (stages I and II, n = 39) and late‐stage PBC (stages III and IV, n = 27). Emperipolesis was measured in liver sections stained with haematoxylin‐eosin. The expressions of CK19, CD3, CD4, CD8, CD20, Ki67 and apoptosis of BECs were evaluated by immunohistochemistry or immunofluorescence double labelling. Emperipolesis was observed in 62.1% of patients with PBC, and BECs were predominantly host cells. The number of infiltrating CD3+ and CD8+ T cells correlated with the advancement of emperipolesis (R2 = 0.318, P < .001; R2 = 0.060, P < .05). The cell numbers of TUNEL‐positive BECs and double staining for CK19 and Ki67 showed a significant positive correlation with emperipolesis degree (R2 = 0.236, P < .001; R2 = 0.267, P < .001). We conclude that emperipolesis mediated by CD8+ T cells appears to be relevant to apoptosis of BEC and thus may aggravate the further injury of interlobular bile ducts.

show abstract

Fluorescence-Activated Cell Sorting Analysis of Heterotypic Cell-in-Cell Structures

Cited by 16 publications

References 23 publications

Detecting cell-in-cell structures in human tumor samples by E-cadherin/CD68/CD45 triple staining

Detecting cell-in-cell structures in human tumor samples by E-cadherin/CD68/CD45 triple staining

Impaired formation of homotypic cell-in-cell structures in human tumor cells lacking alpha-catenin expression

Emperipolesis mediated by CD8⁺ T cells correlates with biliary epithelia cell injury in primary biliary cholangitis

Contact Info

Product

Resources

About

Fluorescence-Activated Cell Sorting Analysis of Heterotypic Cell-in-Cell Structures

Cited by 16 publications

References 23 publications

Detecting cell-in-cell structures in human tumor samples by E-cadherin/CD68/CD45 triple staining

Detecting cell-in-cell structures in human tumor samples by E-cadherin/CD68/CD45 triple staining

Impaired formation of homotypic cell-in-cell structures in human tumor cells lacking alpha-catenin expression

Emperipolesis mediated by CD8+ T cells correlates with biliary epithelia cell injury in primary biliary cholangitis

Contact Info

Product

Resources

About

Emperipolesis mediated by CD8⁺ T cells correlates with biliary epithelia cell injury in primary biliary cholangitis