Establishment and characterization of usm-bc-1 cell line derived from human urinary bladder cancer
Transitional cell carcinoma of the urinary bladder is one of the common diseases that causes morbidity and mortality worldwide. In Malaysia, it lines in the 10 most frequent cancer occurred among males. Since most of the current urinary bladder cancer cell lines were established from populations in...
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| Main Author: | |
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| Format: | Thesis |
| Language: | English |
| Published: |
2013
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| Subjects: | |
| Online Access: | http://eprints.usm.my/60901/1/TEO%20PEY%20SZE%20-%20e.pdf |
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| Summary: | Transitional cell carcinoma of the urinary bladder is one of the common diseases that causes morbidity and mortality worldwide. In Malaysia, it lines in the 10 most frequent cancer occurred among males. Since most of the current urinary bladder cancer cell lines were established from populations in western countries while significant genotypic variations have been reported among various populations, this project aimed at establishing
the first cell line for providing an ideal cancer cell model for population in Malaysia and
countries nearby acquires significance. The urinary bladder cancer cells were isolated from
bladder cancer tissues obtained from a Malay patient and the cells obtained were cultured
continuously over 30 passages ( > 2 years). The cell line was characterized by observing its
morphology, growth characteristics, presence of cytokeratin 7 and desmin markers,
expression of Ki-67, vascular endothelial growth factor (VEGF) as well as p21 proteins.
Chromosome analysis also was performed to further characterize the cell line designated as
USM-BC-1. The authenticity of the obtained cells was investigated using Short Tandem
Repeat (STR) analysis and contamination by mycoplasma species was ascertained. Results
revealed that the USM-BC-1 cell line was composed of predominantly elongated spindle
shape cells with clearly defined margins and no mycoplasma was found contaminating the
established cell line. The population of monolayer cells doubled at about 44.6 hours and
exhibited good viability when grown from cryopreserved stocks. Positive staining for
cytokeratin 7 confirmed the identity of the urothelial cells while the negative staining for
desmin indicated the absence of fibroblast. Ki-67 (345 kDa) was highly expressed in the
USM-BC-1 cell line indicating high proliferation rate of USM-BC-1 cell. Meanwhile, both VEGF and p21 expressions remained low indicating the possible role of other
angiostimulatory proteins and the USM-BC-1 cell line possess neoplastic behavior,
respectively. Chromosomal study proved the occurance of genetic instability when the cell
line exhibited unstable chromosome numbers ranged from 44-57 with a modal number of
44, abnormal chromosomes structure and no Y chromosome observed in 80% of the
metaphase cells. The high similarity (96 %) in the sequence of the STR observed between
the donor and the newly established cell line further confirmed its authenticity. Hence, this
newly established cell line (USM-BC-1) that has been well characterized may prove useful
in cancer research for developing future diagnostic and therapeutic targets, taking into
consideration the genotypic variations among Asian population. |
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