Genetic Heterogeneity of Single Circulating Tumour Cells in Colorectal Carcinoma
The aim of the present study was to isolate and investigate the genetic heterogeneities in single circulating tumour cells (CTCs) from patients with colorectal carcinoma (CRC). Twenty-eight single CTCs were collected from eight patients with CRC using a negative immunomagnetic enrichment method. Aft...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2020-10-01
|
| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/21/20/7766 |
| _version_ | 1797550367417827328 |
|---|---|
| author | Faysal Bin Hamid Vinod Gopalan Marco Matos Cu-Tai Lu Alfred King-yin Lam |
| author_facet | Faysal Bin Hamid Vinod Gopalan Marco Matos Cu-Tai Lu Alfred King-yin Lam |
| author_sort | Faysal Bin Hamid |
| collection | DOAJ |
| description | The aim of the present study was to isolate and investigate the genetic heterogeneities in single circulating tumour cells (CTCs) from patients with colorectal carcinoma (CRC). Twenty-eight single CTCs were collected from eight patients with CRC using a negative immunomagnetic enrichment method. After validation with glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene expression in 3 colon cancer cell lines, a panel of 19 genes were used to analyse the single CTCs (<i>n</i> = 28), primary colorectal carcinoma tissues (<i>n</i> = 8) and colon carcinoma cells (<i>n</i> = 6) using real-time qPCR. Genetic heterogeneities were assessed by comparing gene expression profiles of single CTCs from the different patients and in the same patient, respectively. Genetic profiling of the single CTCs showed extensive heterogeneities of the selected genes among the CTCs. Hierarchical clustering analyses exhibited two clusters of CTCs with differentially expressed genes, which highlighted different modifications from the primary carcinomas. Further, the genetic heterogeneities were observed between different patients or in the same patient. Finally, <i>AKT1</i> expression was significantly (<i>p</i> = 0.0129) higher in single CTCs from CRC of advanced pathological stages (III or IV) CRC than in CTCs from CRC of early stages (I or II). Our findings suggest that single-cell genetic analysis can monitor the genetic heterogeneities and guide the personalised therapeutic targets in clinical sectors. |
| first_indexed | 2024-03-10T15:28:17Z |
| format | Article |
| id | doaj.art-1481f6d52a374c61a7e4ab1ab8fbc2f1 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T15:28:17Z |
| publishDate | 2020-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-1481f6d52a374c61a7e4ab1ab8fbc2f12023-11-20T17:50:32ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672020-10-012120776610.3390/ijms21207766Genetic Heterogeneity of Single Circulating Tumour Cells in Colorectal CarcinomaFaysal Bin Hamid0Vinod Gopalan1Marco Matos2Cu-Tai Lu3Alfred King-yin Lam4Cancer Molecular Pathology, School of Medicine, Griffith University, Gold Coast, QLD 4222, AustraliaCancer Molecular Pathology, School of Medicine, Griffith University, Gold Coast, QLD 4222, AustraliaOncology, Gold Coast University Hospital, Gold Coast, QLD 4215, AustraliaColorectal Surgery, Gold Coast University Hospital, Gold Coast, QLD 4215, AustraliaCancer Molecular Pathology, School of Medicine, Griffith University, Gold Coast, QLD 4222, AustraliaThe aim of the present study was to isolate and investigate the genetic heterogeneities in single circulating tumour cells (CTCs) from patients with colorectal carcinoma (CRC). Twenty-eight single CTCs were collected from eight patients with CRC using a negative immunomagnetic enrichment method. After validation with glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene expression in 3 colon cancer cell lines, a panel of 19 genes were used to analyse the single CTCs (<i>n</i> = 28), primary colorectal carcinoma tissues (<i>n</i> = 8) and colon carcinoma cells (<i>n</i> = 6) using real-time qPCR. Genetic heterogeneities were assessed by comparing gene expression profiles of single CTCs from the different patients and in the same patient, respectively. Genetic profiling of the single CTCs showed extensive heterogeneities of the selected genes among the CTCs. Hierarchical clustering analyses exhibited two clusters of CTCs with differentially expressed genes, which highlighted different modifications from the primary carcinomas. Further, the genetic heterogeneities were observed between different patients or in the same patient. Finally, <i>AKT1</i> expression was significantly (<i>p</i> = 0.0129) higher in single CTCs from CRC of advanced pathological stages (III or IV) CRC than in CTCs from CRC of early stages (I or II). Our findings suggest that single-cell genetic analysis can monitor the genetic heterogeneities and guide the personalised therapeutic targets in clinical sectors.https://www.mdpi.com/1422-0067/21/20/7766CTCsingle-cell analysisgene expressionheterogeneitycolorectal carcinoma |
| spellingShingle | Faysal Bin Hamid Vinod Gopalan Marco Matos Cu-Tai Lu Alfred King-yin Lam Genetic Heterogeneity of Single Circulating Tumour Cells in Colorectal Carcinoma International Journal of Molecular Sciences CTC single-cell analysis gene expression heterogeneity colorectal carcinoma |
| title | Genetic Heterogeneity of Single Circulating Tumour Cells in Colorectal Carcinoma |
| title_full | Genetic Heterogeneity of Single Circulating Tumour Cells in Colorectal Carcinoma |
| title_fullStr | Genetic Heterogeneity of Single Circulating Tumour Cells in Colorectal Carcinoma |
| title_full_unstemmed | Genetic Heterogeneity of Single Circulating Tumour Cells in Colorectal Carcinoma |
| title_short | Genetic Heterogeneity of Single Circulating Tumour Cells in Colorectal Carcinoma |
| title_sort | genetic heterogeneity of single circulating tumour cells in colorectal carcinoma |
| topic | CTC single-cell analysis gene expression heterogeneity colorectal carcinoma |
| url | https://www.mdpi.com/1422-0067/21/20/7766 |
| work_keys_str_mv | AT faysalbinhamid geneticheterogeneityofsinglecirculatingtumourcellsincolorectalcarcinoma AT vinodgopalan geneticheterogeneityofsinglecirculatingtumourcellsincolorectalcarcinoma AT marcomatos geneticheterogeneityofsinglecirculatingtumourcellsincolorectalcarcinoma AT cutailu geneticheterogeneityofsinglecirculatingtumourcellsincolorectalcarcinoma AT alfredkingyinlam geneticheterogeneityofsinglecirculatingtumourcellsincolorectalcarcinoma |