Combination of laser microdissection, 2D-DIGE and MALDI-TOF MS to identify protein biomarkers to predict colorectal cancer spread
Abstract Biomarkers are urgently required to support current histological staging to provide additional accuracy in stratifying colorectal cancer (CRC) patients according to risk of spread to properly assign adjuvant chemotherapy after surgery. Chemotherapy is given to patients with stage III to red...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2019-01-01
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| Series: | Clinical Proteomics |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12014-019-9223-7 |
| _version_ | 1819264110133510144 |
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| author | Chandra Kirana Lifeng Peng Rose Miller John P. Keating Corinne Glenn Hongjun Shi T. William Jordan Guy J. Maddern Richard S. Stubbs |
| author_facet | Chandra Kirana Lifeng Peng Rose Miller John P. Keating Corinne Glenn Hongjun Shi T. William Jordan Guy J. Maddern Richard S. Stubbs |
| author_sort | Chandra Kirana |
| collection | DOAJ |
| description | Abstract Biomarkers are urgently required to support current histological staging to provide additional accuracy in stratifying colorectal cancer (CRC) patients according to risk of spread to properly assign adjuvant chemotherapy after surgery. Chemotherapy is given to patients with stage III to reduce the risk of recurrence but is controversial in stage II patients. Up to 25% of stage II patients will relapse within 5 years after tumor removal and when this occurs cure is seldom possible. The aim of this study was to identify protein biomarkers to stratify risk of spread of CRC patients. Laser micro-dissection was used to isolate cancer cells from primary colorectal tumors of stage II patients which did or did not metastasize within 5 years after surgical resection. Protein expression differences between two groups of tumors were profiled by 2D-DIGE with saturation CyDye labeling and identified using MALDI-TOF mass spectrometry. Evaluation of protein candidates was conducted using tissue micro array (TMA) immunohistochemistry on 125 colorectal tumor tissue samples of different stages. A total of 55 differentially expressed proteins were identified. Ten protein biomarkers were chosen based on p value and ratio between non metastasized and metastazised groups and evaluated on 125 tissues using TMA immunohistochemistry. Expression of HLAB, protein 14-3-3β, LTBP3, ADAMTS2, JAG2 and NME2 on tumour cells was significantly associated with clinical parameters related to tumour progression, invasion and metastasis. Kaplan–Meier survival curve showed strong expression of six proteins was associated with good CRC specific survival. Expression of HLAB, ADAMTS2, LTBP3, JAG2 and NME2 on tumour cells, was associated with tumour progression and invasion, metastasis and CRC specific survival may serve as potential biomarkers to stratify CRC patients into low and high risk of tumour metastasis. Combined methods of laser microdissection, 2D DIGE with saturation labelling and MALDI-TOF MS proved to be resourceful techniques capable of identifying protein biomarkers to predict risk of spread of CRC to liver. |
| first_indexed | 2024-12-23T20:24:16Z |
| format | Article |
| id | doaj.art-40861744a52549cebad42bef6a7a48de |
| institution | Directory Open Access Journal |
| issn | 1542-6416 1559-0275 |
| language | English |
| last_indexed | 2024-12-23T20:24:16Z |
| publishDate | 2019-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Clinical Proteomics |
| spelling | doaj.art-40861744a52549cebad42bef6a7a48de2022-12-21T17:32:26ZengBMCClinical Proteomics1542-64161559-02752019-01-0116111310.1186/s12014-019-9223-7Combination of laser microdissection, 2D-DIGE and MALDI-TOF MS to identify protein biomarkers to predict colorectal cancer spreadChandra Kirana0Lifeng Peng1Rose Miller2John P. Keating3Corinne Glenn4Hongjun Shi5T. William Jordan6Guy J. Maddern7Richard S. Stubbs8Discipline of Surgery, The Queen Elizabeth Hospital, Basil Hetzel Research Institute, University of AdelaideCentre for Biodiscovery and School of Biological Sciences, Victoria University of WellingtonDepartment of Pathology and Molecular Medicine, Otago University of WellingtonCoastal and Coast District Health Board, Department of Surgery, Wellington HospitalCoastal and Coast District Health Board, Department of Surgery, Wellington HospitalWakefield Biomedical Research Unit, Wakefield Clinic, Wakefield HospitalCentre for Biodiscovery and School of Biological Sciences, Victoria University of WellingtonDiscipline of Surgery, The Queen Elizabeth Hospital, Basil Hetzel Research Institute, University of AdelaideWakefield Biomedical Research Unit, Wakefield Clinic, Wakefield HospitalAbstract Biomarkers are urgently required to support current histological staging to provide additional accuracy in stratifying colorectal cancer (CRC) patients according to risk of spread to properly assign adjuvant chemotherapy after surgery. Chemotherapy is given to patients with stage III to reduce the risk of recurrence but is controversial in stage II patients. Up to 25% of stage II patients will relapse within 5 years after tumor removal and when this occurs cure is seldom possible. The aim of this study was to identify protein biomarkers to stratify risk of spread of CRC patients. Laser micro-dissection was used to isolate cancer cells from primary colorectal tumors of stage II patients which did or did not metastasize within 5 years after surgical resection. Protein expression differences between two groups of tumors were profiled by 2D-DIGE with saturation CyDye labeling and identified using MALDI-TOF mass spectrometry. Evaluation of protein candidates was conducted using tissue micro array (TMA) immunohistochemistry on 125 colorectal tumor tissue samples of different stages. A total of 55 differentially expressed proteins were identified. Ten protein biomarkers were chosen based on p value and ratio between non metastasized and metastazised groups and evaluated on 125 tissues using TMA immunohistochemistry. Expression of HLAB, protein 14-3-3β, LTBP3, ADAMTS2, JAG2 and NME2 on tumour cells was significantly associated with clinical parameters related to tumour progression, invasion and metastasis. Kaplan–Meier survival curve showed strong expression of six proteins was associated with good CRC specific survival. Expression of HLAB, ADAMTS2, LTBP3, JAG2 and NME2 on tumour cells, was associated with tumour progression and invasion, metastasis and CRC specific survival may serve as potential biomarkers to stratify CRC patients into low and high risk of tumour metastasis. Combined methods of laser microdissection, 2D DIGE with saturation labelling and MALDI-TOF MS proved to be resourceful techniques capable of identifying protein biomarkers to predict risk of spread of CRC to liver.http://link.springer.com/article/10.1186/s12014-019-9223-7Colorectal cancerPrognosisBiomarkersProteomicsLiver metastasis |
| spellingShingle | Chandra Kirana Lifeng Peng Rose Miller John P. Keating Corinne Glenn Hongjun Shi T. William Jordan Guy J. Maddern Richard S. Stubbs Combination of laser microdissection, 2D-DIGE and MALDI-TOF MS to identify protein biomarkers to predict colorectal cancer spread Clinical Proteomics Colorectal cancer Prognosis Biomarkers Proteomics Liver metastasis |
| title | Combination of laser microdissection, 2D-DIGE and MALDI-TOF MS to identify protein biomarkers to predict colorectal cancer spread |
| title_full | Combination of laser microdissection, 2D-DIGE and MALDI-TOF MS to identify protein biomarkers to predict colorectal cancer spread |
| title_fullStr | Combination of laser microdissection, 2D-DIGE and MALDI-TOF MS to identify protein biomarkers to predict colorectal cancer spread |
| title_full_unstemmed | Combination of laser microdissection, 2D-DIGE and MALDI-TOF MS to identify protein biomarkers to predict colorectal cancer spread |
| title_short | Combination of laser microdissection, 2D-DIGE and MALDI-TOF MS to identify protein biomarkers to predict colorectal cancer spread |
| title_sort | combination of laser microdissection 2d dige and maldi tof ms to identify protein biomarkers to predict colorectal cancer spread |
| topic | Colorectal cancer Prognosis Biomarkers Proteomics Liver metastasis |
| url | http://link.springer.com/article/10.1186/s12014-019-9223-7 |
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