Detection of Circulating Tumor Cells Using Three-dimensional and Conditionally Reprogrammed Culture Methods
0
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
Background: Circulating tumor cells (CTCs) are rare in the circulation of patients with tumors and are very difficult to detect. In this study, we cultured CTCs in a three-dimensionally reprogrammed condition using the peripheral blood of patients with cancer. Methods: We evaluated the sensitivity of a three-dimensional (3D) conditionally reprogrammed cell (CRC) culture method using SNU-484 gastric cancer cells. From April 2017 to January 2018, blood was collected from 30 patients diagnosed with colorectal or breast cancer at the Daegu Catholic University Medical Center. After lysis of the red blood cells, the blood samples were centrifuged to isolate the mononuclear cells. We used Matrigel to create 3D conditions and reprogrammed the isolated cells using a Rho-kinase inhibitor and J2 feeder cells. After 30 days of culture, the cells were collected and RNA was extracted and amplified for the expression of GAPD, hTERT, and MAGE A1-6 using real-time PCR. Results: The SNU-484 cells were cultured from three cells using the 3D CRC culture method. The cultured blood samples showed some cells or few colonies. The positive rates of hTERT and MAGE A1-6 mRNA were 50% and 30%, respectively. The positive rates of both mRNAs were 60% and 20%, respectively. MAGE and both mRNAs showed significantly increased odds ratios at the tumor, nodes, and metastases (TNM), N, and M stages in all subjects. Conclusions: As 60% of CTCs were detected in the blood of 30 patients with cancer, the 3D CRC culture method was very effective for CTC culture.
Abstract
Background: Circulating tumor cells (CTCs) are rare in the circulation of patients with tumors and are very difficult to detect. In this study, we cultured CTCs in a three-dimensionally reprogrammed condition using the peripheral blood of patients with cancer. Methods: We evaluated the sensitivity of a three-dimensional (3D) conditionally reprogrammed cell (CRC) culture method using SNU-484 gastric cancer cells. From April 2017 to January 2018, blood was collected from 30 patients diagnosed with colorectal or breast cancer at the Daegu Catholic University Medical Center. After lysis of the red blood cells, the blood samples were centrifuged to isolate the mononuclear cells. We used Matrigel to create 3D conditions and reprogrammed the isolated cells using a Rho-kinase inhibitor and J2 feeder cells. After 30 days of culture, the cells were collected and RNA was extracted and amplified for the expression of GAPD, hTERT, and MAGE A1-6 using real-time PCR. Results: The SNU-484 cells were cultured from three cells using the 3D CRC culture method. The cultured blood samples showed some cells or few colonies. The positive rates of hTERT and MAGE A1-6 mRNA were 50% and 30%, respectively. The positive rates of both mRNAs were 60% and 20%, respectively. MAGE and both mRNAs showed significantly increased odds ratios at the tumor, nodes, and metastases (TNM), N, and M stages in all subjects. Conclusions: As 60% of CTCs were detected in the blood of 30 patients with cancer, the 3D CRC culture method was very effective for CTC culture.
참고문헌 (34)
- Ashworth TR, "A case of cancer in which cells similar to those in the tumours were seen in the blood after death", Aust Med J, 14, pp.146~147 (1869)
- Pecot CV, "A novel platform for detection of CK+ and CK-CTCs", Cancer Discov, 1, pp.580~586 (2011)
- Marrinucci D, "Fluid biopsy in patients with metastatic prostate, pancreatic and breast cancers", Phys Biol, 9, pp.016003~ (2012)
- Haber DA, "Blood-based analyses of cancer : circulating tumor cells and circulating tumor DNA", Cancer Discov, 4, pp.650~661 (2014)
- Azim HA, Jr., "Circulating tumor cells and response to neoadjuvant paclitaxel and HER2-targeted therapy: a sub-study from the NeoALTTO phase III trial", Breast, 22, pp.1060~1065 (2013)
- Miller MC, "Significance of circulating tumor cells detected by the CellSearch System in patients with metastatic breast colorectal and prostate cancer", J Oncol, 2010, pp.617421~ (2010)
- Kang Y, "Tumor cell dissemination : emerging biological insights from animal models and cancer patients", Cancer Cell, 23, pp.573~581 (2013)
- Meng S, "Circulating tumor cells in patients with breast cancer dormancy", Clin Cancer Res, 10, pp.8152~8162 (2004)
- Dong Y, "Micro uidics and circulating tumor cells", J Mol Diagn, 15, pp.149~157 (2013)
- Gupta V, "ApoStream™, a new dielectrophoretic device for antibody independent isolation and recovery of viable cancer cells from blood", Biomicro uidics, 6, pp.24133~ (2012)
- Nagrath S, "Isolation of rare circulating tumour cells in cancer patients by microchip technology", Nature, 450, pp.1235~1239 (2007)
- Gleghorn JP, "Capture of circulating tumor cells from whole blood of prostate cancer patients using geometrically enhanced differential immunocapture(GEDI)and a prostate-specific antibody", Lab Chip, 10, pp.27~29 (2010)
- Zhang L, "The identification and characterization of breast cancer CTCs competent for brain metastasis", Sci Transl Med, 5, pp.180ra48~ (2013)
- Yu M, "Ex vivo culture of circulating breast tumor cells for individualized testing of drug susceptibility", Science, 345, pp.216~220 (2014)
- Jeong YJ, "Detection of circulating tumor cells in patients with breast cancer using the conditionally reprogrammed cell culture method and reverse transcription PCR of hTERT and MAGE A1 6", Oncol Lett, 20, pp.78~ (2020)
- Yang CS, "Detection of circulating gastrointestinal cancer cells in conditionally reprogrammed cell culture", In Vivo, 35, pp.1515~1520 (2021)
- Liu X, "ROCK inhibitor and feeder cells induce the conditional reprogramming of epithelial cells", Am J Pathol, 180, pp.599~607 (2012)
- Suprynowicz FA, "Conditionally reprogrammed cells represent a stem-like state of adult epithelial cells", Proc Natl Acad Sci U S A, 109, pp.20035~20040 (2012)
- Benton G, "Multiple uses of basement membrane-like matrix(BME", Matrigel)in vitro and in vivo with cancer cells, 2011, pp.128, 1751~1757
- Liu X, "Utilizing matrigel transwell invasion assay to detect and enumerate circulating tumor cells", Methods Mol Biol, 1634, pp.277~282 (2017)
- Ferreira MM, "Circulating tumor cell technologies", Mol Oncol, 10, pp.374~394 (2016)
- Ilie M, "Expression of MET in circulating tumor cells correlates with expression in tumor tissue from advanced-stage lung cancer patients", Oncotarget, 8, pp.26112~26121 (2017)
- Masuda T, "Clinical and biological significance of circulating tumor cells in cancer", Mol Oncol, 10, pp.408~417 (2016)
- Szilágyi M, "Circulating cell-free nucleic acids : Main characteristics and clinical application", Int J Mol Sci, 21, pp.6827~ (2020)
- Stewart CM, "Circulating cell-free DNA for non-invasive cancer management", Cancer Genet, 228-9, pp.169~179 (2018)
- Jiang T, "Role of circulating-tumor DNA analysis in non-small cell lung cancer", Lung Cancer, 90, pp.128~134 (2015)
- Brock G, "Liquid biopsy for cancer screening, patient stratification and monitoring", Transl Cancer Res, 4, pp.280~290 (2015)
- Kim DD, "Melanoma antigen-encoding gene family member A1-6 and hTERT in the detection of circulating tumor cells following CD45-depletion and RNA extraction", Oncol Lett, 14, pp.837~843 (2017)
- Bobek V, "Circulating tumor cells in pancreatic cancer patients : Enrichment and cultivation", World J Gastroenterol, 20, pp.17163~17170 (2014)
- Rodríguez-Lee M, "Effect of blood collection tube type and time to processing on the enumeration and high-content characterization of circulating tumor cells using the high-definition single-cell assay", Arch Pathol Lab Med, 142, pp.198~207 (2018)
- Cayrefourcq L, "Establishment and characterization of a cell line from human circulating colon cancer cells", Cancer Res, 75, pp.892~901 (2015)
- Kaifi JT, "Circulating tumor cell levels are elevated in colorectal cancer patients with high tumor burden in the liver", Cancer Biol Ther, 16, pp.690~698 (2015)
- Takeda K, "Analysis of colorectal cancer-related mutations by liquid biopsy : Utility of circulating cell-free DNA and circulating tumor cells", Cancer Sci, 110, pp.3497~3509 (2019)
- Bidard FC, "Circulating Tumor Cells and Circulating Tumor DNA Detection in Potentially Resectable Metastatic Colorectal Cancer : A Prospective Ancillary Study to the Unicancer Prodige-14 Trial", Cells, 8, pp.516~ (2019)