Enumeration, characterisation and clinicopathological significance of circulating tumour cells in patients with colorectal carcinoma

Hamid, Faysal Bin; Lu, Cu-Tai; Matos, Marco; Cheng, Tracie; Gopalan, Vinod; Lam, Alfred King‐Yin

doi:10.1016/j.cancergen.2021.02.002

Cited by 4 publications

(5 citation statements)

References 44 publications

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“…The identification of CTCs and their specific gene profiles could offer new perspectives that may improve the prediction of metastasis formation, as well as representing a promising approach for determining better therapeutic targets. Previous studies have reported a significant correlation between CTC counts and pathological stages of various cancers, including CRC [2,18,19,[21][22][23][24][25]. In this study, we noted that late-stage cancers more often had higher CTC counts when compared with early-stage cancers (p = 0.045).…”

Section: Discussionsupporting

confidence: 54%

“…Preliminary experiments had been performed previously, in which different numbers of colon cancer cell lines were added to peripheral blood collected from healthy donors to evaluate the sensitivity and specificity of the CTC enrichment technique (negative selection method) by estimating the recovery rate of CTCs from a 5 mL blood sample [18]. Screening for different subpopulations of CTC was also validated using a panel of six antibodies (EpCAM, CK18, SNAIL1, MMP-9, E-cadherin, and BCL-2) described in detail previously [18]. In this present study, we selected four epithelial and EMT-related markers from the panel of antibodies (EpCAM, SNAIL1, MMP-9 and E-cadherin) to characterise different subpopulations of CTCs in different patients.…”

Section: Enumeration Of Ctcs In Patients With Crcmentioning

confidence: 99%

“…Then, the enriched CTCs were seeded in a 96-well plate (50 µL per well) for immunofluorescence and in a 6-well plate for downstream analysis, followed by overnight incubation. The composition of CTC growth medium was described in a previously published article [18]. Peripheral blood samples from 6 healthy donors were processed as negative controls in the same way as previously performed for patients with CRC.…”

Section: Enrichment and Isolation Of Ctcsmentioning

confidence: 99%

“…Immunofluorescence staining was performed for the identification of CTCs using a cocktail of four primary antibodies for EpCAM (Thermo Fisher Scientific, Waltham, MA, USA), SNAIL1, E-cadherin and MMP-9 (Santa Cruz Biotechnology, CA, USA) as described previously [18]. In brief, the enriched cells were fixed with 100% methanol for 10 min at −20 • C and were permeabilised with 0.2% Triton X-100 for 10 min.…”

Section: Immunofluorescence Stainingmentioning

confidence: 99%

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Gene Expression Analysis of Immune Regulatory Genes in Circulating Tumour Cells and Peripheral Blood Mononuclear Cells in Patients with Colorectal Carcinoma

Aktar

Gamage

Cheng

et al. 2023

IJMS

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Information regarding genetic alterations of driver cancer genes in circulating tumour cells (CTCs) and their surrounding immune microenvironment nowadays can be employed as a real-time monitoring platform for translational applications such as patient response to therapeutic targets, including immunotherapy. This study aimed to investigate the expression profiling of these genes along with immunotherapeutic target molecules in CTCs and peripheral blood mononuclear cells (PBMCs) in patients with colorectal carcinoma (CRC). Expression of p53, APC, KRAS, c-Myc, and immunotherapeutic target molecules PD-L1, CTLA-4, and CD47 in CTCs and PBMCs were analysed by qPCR. Their expression in high versus low CTC-positive patients with CRC was compared and clinicopathological correlations between these patient groups were analysed. CTCs were detected in 61% (38 of 62) of patients with CRC. The presence of higher numbers of CTCs was significantly correlated with advanced cancer stages (p = 0.045) and the subtypes of adenocarcinoma (conventional vs. mucinous, p = 0.019), while being weakly correlated with tumour size (p = 0.051). Patients with lower numbers of CTCs had higher expression of KRAS. Higher KRAS expression in CTCs was negatively correlated with tumour perforation (p = 0.029), lymph node status (p = 0.037), distant metastasis (p = 0.046) and overall staging (p = 0.004). CTLA-4 was highly expressed in both CTCs and PBMCs. In addition, CTLA-4 expression was positively correlated with KRAS (r = 0.6878, p = 0.002) in the enriched CTC fraction. Dysregulation of KRAS in CTCs might evade the immune system by altering the expression of CTLA-4, providing new insights into the selection of therapeutic targets at the onset of the disease. Monitoring CTCs counts, as well as gene expression profiling of PBMCs, can be helpful in predicting tumour progression, patient outcome and treatment.

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Section: Discussionsupporting

confidence: 54%

Section: Enumeration Of Ctcs In Patients With Crcmentioning

confidence: 99%

Section: Enrichment and Isolation Of Ctcsmentioning

confidence: 99%

Section: Immunofluorescence Stainingmentioning

confidence: 99%

See 2 more Smart Citations

Gene Expression Analysis of Immune Regulatory Genes in Circulating Tumour Cells and Peripheral Blood Mononuclear Cells in Patients with Colorectal Carcinoma

Aktar

Gamage

Cheng

et al. 2023

IJMS

Self Cite

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“…CTCs were enriched from 10 mL of peripheral blood according to the manufacturer’s protocol (EasySep TM Direct Human CTC Enrichment Kit, STEMCELL Technologies, Vancouver, BC, Canada). The procedures of CTC enrichment and isolation as well as the cell spiking method to verify the specificity and sensitivity of this method have been previously reported in detail [ 30 , 33 ]. The enriched cell pellets were seeded in a 96-well plate after centrifugation at 450 rcf for 7 min and incubated at 37 °C for 12 h.…”

Section: Methodsmentioning

confidence: 99%

Correlation between KRAS Mutation and CTLA-4 mRNA Expression in Circulating Tumour Cells: Clinical Implications in Colorectal Cancer

Aktar,

Islam,

Cheng

et al. 2023

Genes

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Combination strategies of KRAS inhibition with immunotherapy in treating advanced or recurrent colorectal carcinoma (CRC) may need to be assessed in circulating tumour cells (CTCs) to achieve better clinical outcomes. This study aimed to investigate the genomic variations of KRAS in CTCs and matched CRC tissues and compared mRNA expression of KRAS and CTLA-4 between wild-type and KRAS-mutated CTCs and CRC tissues. Clinicopathological correlations were also compared. Six known mutations of KRAS were identified at both codon 12 and codon 13 (c.35G>T/G12V, c.35G>A7/G12D, c.35G>C/G12A, c.34G>A/G12S, c.38G>C/G13A, and c.38G>A/G13D). Three CTC samples harboured the identified mutations (16.7%; 3/18), while fifteen matched primary tumour tissues (65.2%, 15/23) showed the mutations. CTCs harbouring the KRAS variant were different from matched CRC tissue. All the mutations were heterozygous. Though insignificant, CTLA-4 mRNA expression was higher in patients carrying KRAS mutations. Patients harbouring KRAS mutations in CTCs were more likely to have poorly differentiated tumours (p = 0.039) and with lymph node metastasis (p = 0.027) and perineural invasion (p = 0.014). KRAS mutations in CTCs were also significantly correlated with overall pathological stages (p = 0.027). These findings imply the genetic basis of KRAS with immunotherapeutic target molecules based on a real-time platform. This study also suggests the highly heterogeneous nature of cancer cells, which may facilitate the assessment of clonal dynamics across a single patient’s disease.

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Comparison of genomic profiling of patient-matched primary colorectal and surgical resected distant metastatic (stage IV) colorectal carcinoma for drug actionability

Jan,

Lu,

Lam

2024

Human Pathology

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Enumeration, characterisation and clinicopathological significance of circulating tumour cells in patients with colorectal carcinoma

Cited by 4 publications

References 44 publications

Gene Expression Analysis of Immune Regulatory Genes in Circulating Tumour Cells and Peripheral Blood Mononuclear Cells in Patients with Colorectal Carcinoma

Gene Expression Analysis of Immune Regulatory Genes in Circulating Tumour Cells and Peripheral Blood Mononuclear Cells in Patients with Colorectal Carcinoma

Correlation between KRAS Mutation and CTLA-4 mRNA Expression in Circulating Tumour Cells: Clinical Implications in Colorectal Cancer

Comparison of genomic profiling of patient-matched primary colorectal and surgical resected distant metastatic (stage IV) colorectal carcinoma for drug actionability

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