Characterisation of metabolic changes in colorectal cancer cells of different stages / Hazwani Mohd Yusof
Accurate diagnosis and staging of colorectal cancer (CRC) lead to better prognosis. Identification of molecular biomarkers may prove useful, but there is a lack of data on the molecular pathophysiology of CRC progression. Characterisation of metabolic changes in CRC cells may provide useful tools...
Saved in:
| Main Author: | |
|---|---|
| Format: | Thesis |
| Language: | English |
| Published: |
2020
|
| Subjects: | |
| Online Access: | https://ir.uitm.edu.my/id/eprint/60724/1/60724.pdf |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| id |
my-uitm-ir.60724 |
|---|---|
| record_format |
uketd_dc |
| spelling |
my-uitm-ir.607242022-06-01T02:38:07Z Characterisation of metabolic changes in colorectal cancer cells of different stages / Hazwani Mohd Yusof 2020-04 Mohd Yusof, Hazwani Cancer Accurate diagnosis and staging of colorectal cancer (CRC) lead to better prognosis. Identification of molecular biomarkers may prove useful, but there is a lack of data on the molecular pathophysiology of CRC progression. Characterisation of metabolic changes in CRC cells may provide useful tools for study of potential therapeutic drug actions. This study was aimed to characterise the metabolites profiles in CRC cells of different stages and the metabolic pathways affected. In this study, normal colon cell lines; CCD 840 CoN and CRC cells lines of different stages; SW 1116 (stage A), HT 29 and SW 480 (stage B), HCT 15 and DLD-1 (stage C), and HCT 116 (stage D) were used. The global metabolomics profiling was performed using liquid chromatographymass spectrometry (LC/MS). Mass Profiler Professional and Metaboanalyst software were used for statistical and pathway analysis. METLIN database was used for identification of metabolites. MS/MS was used for validation of detected metabolites. Validation of the affected metabolic pathways in CRC was performed in which expression of SLC6A14 (amino acid transporter), riboflavin kinase and FAD synthetase (enzymes in riboflavin metabolism pathway) were determined using Western blot. Lmethionine and FAD levels, cells viability and rate of apoptosis activity were determined with and without the presence of alpha-methyltryptophan, and lumichrome which are inhibitors of SLC6A14, and riboflavin uptake, respectively. The results showed that there were differential metabolites identified in CRC cells of different stages, which could be used as potential metabolite biomarkers for staging. (R)-1-O- [b-D-glucopyranosyl-(1->6)-b-D-glucopyranoside]-1,3-octanediol, lauroyl diethanolamide, lysoPE(0:0/18:1(11Z)), lysoPE(0:0/22:5(4Z,7Z,10Z,13Z,16Z)), phosphocholine, S-(formylmethyl)glutathione, and N-acetyl-DL-methionine were the most important differential metabolites identified in stage A. In stage B, 1,2,4- nonadecanetriol, dodecanoylcarnitine, hericene B, l-hexanoylcarnitine, pyroglutamic acid and S-furanopetasitin were the differential metabolites while in stage C, the differential metabolites were bis-γ-glutamylcystine, hippuric acid, lumichrome, lysoPE(16:1(9Z)/0:0), PE(22:5(7Z,10Z,13Z,16Z,19Z)/15:0), and purine. FAD, methyhiomethyl butyrate, muzanzagenim, 3b-hydroxy-5-cholenoic acid, ATP, Lglutamate and lysoPE(0:0/20:0) were the differential metabolites identified in CRC cells stage D. The results showed that L-methionine and riboflavin metabolisms were the most prominent pathways affected in CRC cells as the cancer advances. The results also showed that increased level of methionine observed in metabolomics study is probably due to increased expression of SLC6A14. However, expressions of riboflavin kinase and FAD synthetase in CRC were similar to normal cells and hence could not account for the increased level of FMN and FAD as observed in the metabolomics study. The results also showed that metabolites profiles identified in intracellular and secreted CRC cells were different. In conclusion, this study identified several novel differential metabolites which can be used as potential biomarkers for staging. In this study, for the first time, profiles of the intracellular as well as the secreted metabolites were determined which provides a more complete picture of the metabolic derangement that occurs in the development and progression of CRC. The affected metabolic pathways identified in this study may lead to further understanding of the pathophysiology of CRC and hence, its application for treatment strategies. 2020-04 Thesis https://ir.uitm.edu.my/id/eprint/60724/ https://ir.uitm.edu.my/id/eprint/60724/1/60724.pdf text en public phd doctoral Universiti Teknologi MARA Faculty of Medicine Mazlan, Musalmah (Professor Dr.) |
| institution |
Universiti Teknologi MARA |
| collection |
UiTM Institutional Repository |
| language |
English |
| advisor |
Mazlan, Musalmah (Professor Dr.) |
| topic |
Cancer |
| spellingShingle |
Cancer Mohd Yusof, Hazwani Characterisation of metabolic changes in colorectal cancer cells of different stages / Hazwani Mohd Yusof |
| description |
Accurate diagnosis and staging of colorectal cancer (CRC) lead to better prognosis.
Identification of molecular biomarkers may prove useful, but there is a lack of data on
the molecular pathophysiology of CRC progression. Characterisation of metabolic
changes in CRC cells may provide useful tools for study of potential therapeutic drug
actions. This study was aimed to characterise the metabolites profiles in CRC cells of
different stages and the metabolic pathways affected. In this study, normal colon cell
lines; CCD 840 CoN and CRC cells lines of different stages; SW 1116 (stage A), HT
29 and SW 480 (stage B), HCT 15 and DLD-1 (stage C), and HCT 116 (stage D) were
used. The global metabolomics profiling was performed using liquid chromatographymass
spectrometry (LC/MS). Mass Profiler Professional and Metaboanalyst software
were used for statistical and pathway analysis. METLIN database was used for
identification of metabolites. MS/MS was used for validation of detected metabolites.
Validation of the affected metabolic pathways in CRC was performed in which
expression of SLC6A14 (amino acid transporter), riboflavin kinase and FAD synthetase
(enzymes in riboflavin metabolism pathway) were determined using Western blot. Lmethionine
and FAD levels, cells viability and rate of apoptosis activity were
determined with and without the presence of alpha-methyltryptophan, and lumichrome
which are inhibitors of SLC6A14, and riboflavin uptake, respectively. The results
showed that there were differential metabolites identified in CRC cells of different
stages, which could be used as potential metabolite biomarkers for staging. (R)-1-O-
[b-D-glucopyranosyl-(1->6)-b-D-glucopyranoside]-1,3-octanediol, lauroyl
diethanolamide, lysoPE(0:0/18:1(11Z)), lysoPE(0:0/22:5(4Z,7Z,10Z,13Z,16Z)),
phosphocholine, S-(formylmethyl)glutathione, and N-acetyl-DL-methionine were the
most important differential metabolites identified in stage A. In stage B, 1,2,4-
nonadecanetriol, dodecanoylcarnitine, hericene B, l-hexanoylcarnitine, pyroglutamic
acid and S-furanopetasitin were the differential metabolites while in stage C, the
differential metabolites were bis-γ-glutamylcystine, hippuric acid, lumichrome,
lysoPE(16:1(9Z)/0:0), PE(22:5(7Z,10Z,13Z,16Z,19Z)/15:0), and purine. FAD,
methyhiomethyl butyrate, muzanzagenim, 3b-hydroxy-5-cholenoic acid, ATP, Lglutamate
and lysoPE(0:0/20:0) were the differential metabolites identified in CRC
cells stage D. The results showed that L-methionine and riboflavin metabolisms were
the most prominent pathways affected in CRC cells as the cancer advances. The results
also showed that increased level of methionine observed in metabolomics study is
probably due to increased expression of SLC6A14. However, expressions of riboflavin
kinase and FAD synthetase in CRC were similar to normal cells and hence could not
account for the increased level of FMN and FAD as observed in the metabolomics study.
The results also showed that metabolites profiles identified in intracellular and secreted
CRC cells were different. In conclusion, this study identified several novel differential
metabolites which can be used as potential biomarkers for staging. In this study, for the
first time, profiles of the intracellular as well as the secreted metabolites were
determined which provides a more complete picture of the metabolic derangement that
occurs in the development and progression of CRC. The affected metabolic pathways
identified in this study may lead to further understanding of the pathophysiology of
CRC and hence, its application for treatment strategies. |
| format |
Thesis |
| qualification_name |
Doctor of Philosophy (PhD.) |
| qualification_level |
Doctorate |
| author |
Mohd Yusof, Hazwani |
| author_facet |
Mohd Yusof, Hazwani |
| author_sort |
Mohd Yusof, Hazwani |
| title |
Characterisation of metabolic changes in colorectal cancer cells of different stages / Hazwani Mohd Yusof |
| title_short |
Characterisation of metabolic changes in colorectal cancer cells of different stages / Hazwani Mohd Yusof |
| title_full |
Characterisation of metabolic changes in colorectal cancer cells of different stages / Hazwani Mohd Yusof |
| title_fullStr |
Characterisation of metabolic changes in colorectal cancer cells of different stages / Hazwani Mohd Yusof |
| title_full_unstemmed |
Characterisation of metabolic changes in colorectal cancer cells of different stages / Hazwani Mohd Yusof |
| title_sort |
characterisation of metabolic changes in colorectal cancer cells of different stages / hazwani mohd yusof |
| granting_institution |
Universiti Teknologi MARA |
| granting_department |
Faculty of Medicine |
| publishDate |
2020 |
| url |
https://ir.uitm.edu.my/id/eprint/60724/1/60724.pdf |
| _version_ |
1783735161282625536 |