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Clinical significance of phenotyping and karyotyping of circulating tumor cells in patients with advanced gastric cancer.

Li Y, Zhang X, Ge S, Gao J, Gong J, Lu M, Zhang Q, Cao Y, Wang DD, Lin PP, Shen L - Oncotarget (2014)

Bottom Line: Phenotyping of CTCs in HER2 positive AGC patients demonstrated that HER2⁺ CTCs could be effectively eliminated in response to HER2-targeted therapy.Examination of the copy number of chromosome 8 in CTCs provides a potential approach for predicting chemotherapeutic efficacy and monitoring chemo-resistance.Phenotyping and karyotyping of the enriched CTCs upon ploidy of chromosome 8 or HER2 expression is of clinical potential for monitoring chemo-resistance and evaluating therapeutic efficacy for AGC patients.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education), Department of GI Oncology, Peking University Cancer Hospital & Institute, Beijing 100142, China.

ABSTRACT

Background: Karyotyping and phenotyping of circulating tumor cells (CTCs) in therapeutic cancer patients is of particular clinical significance in terms of both identifying chemo-resistant CTC subtypes and understanding CTC evolution.

Methods: The integrated subtraction enrichment (SET) and immunostaining-fluorescence in situ hybridization (iFISH) platform was applied to detect and characterize CTCs in patients with advanced gastric cancer (AGC). Status of human epidermal growth factor receptor 2 (HER2) expressing and aneuploidy of chromosome 8 in CTCs enriched from the patients was examined by SET-iFISH following clinical chemotherapy or HER2-targeted therapy. CellSearch system was applied as a reference control.

Results: Phenotyping of CTCs in HER2 positive AGC patients demonstrated that HER2⁺ CTCs could be effectively eliminated in response to HER2-targeted therapy. Karytotyping of CTCs indicated that distinct CTCs with different ploidies of chromosomes 8 in AGC patients correlated to either sensitivity or resistance of paclitaxel or cisplatin-based chemotherapy. Examination of the copy number of chromosome 8 in CTCs provides a potential approach for predicting chemotherapeutic efficacy and monitoring chemo-resistance.

Conclusions: Phenotyping and karyotyping of the enriched CTCs upon ploidy of chromosome 8 or HER2 expression is of clinical potential for monitoring chemo-resistance and evaluating therapeutic efficacy for AGC patients.

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Related in: MedlinePlus

Detection of HER2 expression status on CTCs(A) Quantification of immnuofluorescence stained HER2 on cell lines performed by SET-IF and CellSearch. HER2 expression was quantified by Alexa 488-conjugated anti-HER2 (green) for SET-IF and FITC labeled anti-HER2 for CellSearch. Scale bar = 5 μm. (B) Detection of HER2 expression on CTCs enriched from pathological HER2+ AGC patients by SET-IF or CellSearch. (C) Changes of total CTCs and HER2+ CTCs following HER2-targeted therapy.
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Figure 2: Detection of HER2 expression status on CTCs(A) Quantification of immnuofluorescence stained HER2 on cell lines performed by SET-IF and CellSearch. HER2 expression was quantified by Alexa 488-conjugated anti-HER2 (green) for SET-IF and FITC labeled anti-HER2 for CellSearch. Scale bar = 5 μm. (B) Detection of HER2 expression on CTCs enriched from pathological HER2+ AGC patients by SET-IF or CellSearch. (C) Changes of total CTCs and HER2+ CTCs following HER2-targeted therapy.

Mentions: Status of HER2 expression on CTCs was further examined by both SET-IF and CellSearch. To quantify HER2 expression on CTCs, a panel of cell lines with different expression levels of HER2 was examined by both SET-IF and CellSearch. Immunofluorescence intensity of HER2 corresponding to the known HER2 expression status serves as the reference standard for quantification of HER2 status on CTCs [21]. As shown in Figure 2A, immunofluorescence intensity of HER2 was scored as 0 for MCF-7 (none expression), 1+ for BT20 (low), 2+ for MDA-MB-453 (medium), and 3+ for SK-BR-3 (high) cells, respectively. In the case of CTCs, 0 and 1+ were defined as HER2−, 2+ and 3+ were classified as HER2+.


Clinical significance of phenotyping and karyotyping of circulating tumor cells in patients with advanced gastric cancer.

Li Y, Zhang X, Ge S, Gao J, Gong J, Lu M, Zhang Q, Cao Y, Wang DD, Lin PP, Shen L - Oncotarget (2014)

Detection of HER2 expression status on CTCs(A) Quantification of immnuofluorescence stained HER2 on cell lines performed by SET-IF and CellSearch. HER2 expression was quantified by Alexa 488-conjugated anti-HER2 (green) for SET-IF and FITC labeled anti-HER2 for CellSearch. Scale bar = 5 μm. (B) Detection of HER2 expression on CTCs enriched from pathological HER2+ AGC patients by SET-IF or CellSearch. (C) Changes of total CTCs and HER2+ CTCs following HER2-targeted therapy.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4196148&req=5

Figure 2: Detection of HER2 expression status on CTCs(A) Quantification of immnuofluorescence stained HER2 on cell lines performed by SET-IF and CellSearch. HER2 expression was quantified by Alexa 488-conjugated anti-HER2 (green) for SET-IF and FITC labeled anti-HER2 for CellSearch. Scale bar = 5 μm. (B) Detection of HER2 expression on CTCs enriched from pathological HER2+ AGC patients by SET-IF or CellSearch. (C) Changes of total CTCs and HER2+ CTCs following HER2-targeted therapy.
Mentions: Status of HER2 expression on CTCs was further examined by both SET-IF and CellSearch. To quantify HER2 expression on CTCs, a panel of cell lines with different expression levels of HER2 was examined by both SET-IF and CellSearch. Immunofluorescence intensity of HER2 corresponding to the known HER2 expression status serves as the reference standard for quantification of HER2 status on CTCs [21]. As shown in Figure 2A, immunofluorescence intensity of HER2 was scored as 0 for MCF-7 (none expression), 1+ for BT20 (low), 2+ for MDA-MB-453 (medium), and 3+ for SK-BR-3 (high) cells, respectively. In the case of CTCs, 0 and 1+ were defined as HER2−, 2+ and 3+ were classified as HER2+.

Bottom Line: Phenotyping of CTCs in HER2 positive AGC patients demonstrated that HER2⁺ CTCs could be effectively eliminated in response to HER2-targeted therapy.Examination of the copy number of chromosome 8 in CTCs provides a potential approach for predicting chemotherapeutic efficacy and monitoring chemo-resistance.Phenotyping and karyotyping of the enriched CTCs upon ploidy of chromosome 8 or HER2 expression is of clinical potential for monitoring chemo-resistance and evaluating therapeutic efficacy for AGC patients.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education), Department of GI Oncology, Peking University Cancer Hospital & Institute, Beijing 100142, China.

ABSTRACT

Background: Karyotyping and phenotyping of circulating tumor cells (CTCs) in therapeutic cancer patients is of particular clinical significance in terms of both identifying chemo-resistant CTC subtypes and understanding CTC evolution.

Methods: The integrated subtraction enrichment (SET) and immunostaining-fluorescence in situ hybridization (iFISH) platform was applied to detect and characterize CTCs in patients with advanced gastric cancer (AGC). Status of human epidermal growth factor receptor 2 (HER2) expressing and aneuploidy of chromosome 8 in CTCs enriched from the patients was examined by SET-iFISH following clinical chemotherapy or HER2-targeted therapy. CellSearch system was applied as a reference control.

Results: Phenotyping of CTCs in HER2 positive AGC patients demonstrated that HER2⁺ CTCs could be effectively eliminated in response to HER2-targeted therapy. Karytotyping of CTCs indicated that distinct CTCs with different ploidies of chromosomes 8 in AGC patients correlated to either sensitivity or resistance of paclitaxel or cisplatin-based chemotherapy. Examination of the copy number of chromosome 8 in CTCs provides a potential approach for predicting chemotherapeutic efficacy and monitoring chemo-resistance.

Conclusions: Phenotyping and karyotyping of the enriched CTCs upon ploidy of chromosome 8 or HER2 expression is of clinical potential for monitoring chemo-resistance and evaluating therapeutic efficacy for AGC patients.

Show MeSH
Related in: MedlinePlus