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Cisplatin‐induced apoptosis in HeLa cells to determine potential mechanisms for drug resistance
Author(s) -
Shipley Mackenzie M.,
Piefer Andrew J.
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.545.1
Subject(s) - apoptosis , gene knockdown , hela , cisplatin , programmed cell death , microbiology and biotechnology , caspase , biology , cancer cell , small interfering rna , intrinsic apoptosis , dna damage , cell , chemistry , cell culture , dna , transfection , cancer , biochemistry , genetics , chemotherapy
The purpose of this project is to study the cisplatin‐induced apoptosis mechanism in HeLa cells. Apoptosis or programmed cell death occurs when molecular lesions are formed in genomic DNA. By using small‐interfering RNA (siRNA), the oncogene c‐Abl will be targeted and knocked down to suppress protein synthesis. c‐Abl encodes a cytoplasmic and nuclear protein tyrosine kinase. This oncogene gets translocated to the mitochondrial membrane and when DNA damage occurs, cytochrome c is released into the cytosol where eventual intrinsic apoptosis occurs. A colorimetric assay kit will be used to measure the amount of caspase‐3 present in cells before and after knockdown of c‐Abl. Caspase‐3 is a cysteine protease with specificity for aspartate residues. All caspases play an important role in the execution‐phase of intrinsic apoptosis. Trypan Blue will also be used to measure the total amount of cell death before and after knockdown of the gene target. siRNA knockdown of c‐Abl should result in decreased caspase‐3 and increased cell survival. The results of this research will determine if c‐Abl is involved in cisplatin mediated apoptosis in a cervical cancer cell line and may reveal potential drug resistance mechanisms. This research is funded by the Hartwick College Department of Chemistry.