Naslov
Comparision of transfection efficiency and antitumour effects of apoptin between different tumour cell lines

Autori
Lemmens, Steffi

Vrsta, podvrsta i kategorija rada
Ocjenski radovi, diplomski rad, preddiplomski

Fakultet
University College Ghent

Mjesto
Ghent, Belgija

Datum
06.06

Godina
2016

Stranica
61

Mentor
Ester, Katja

Ključne riječi
Cancer therapy ; PEI ; HMLE cells ; apoptin ; transient transfection

Sažetak
Cancer stem cells (CSCs) represent a subpopulation of cancer cells that are thought to be responsible for tumour formation, relapse and metastasis. There isn’t any of data of the effect of apoptin on cancer stem cells. These cells are very similar of embryonic stem cells. Both of them have the same working of pluripotency and self-renewal. Cancer stem cells are resistant to a whole range of anti-cancer drugs with different cellular targets. Mechanisms underlying the therapeutic resistance of breast cancer stem cells have also not been fully elucidated, although it has been previously shown that breast cancer stem cells are able to evade apoptosis. In previous experiments, delivery of apoptin using adenoviral vectors resulted in insufficient transfection efficiency in breast CSC model. The breast CSC model consists of two isogenic epithelial breast cell lines, HMLE pBp and HMLE Twist, with latter the showing markers and functional features of cancer stem cells. In order to evaluate potential cytotoxic effect of apoptin toward CSCs, it is important to obtain satisfactory transient transfection efficiency. In the study described in this thesis, we will investigate the (PEI)-plasmid DNA complex as a tool for apoptin delivery into tumour and HMLE cells. Plasmid encoding apoptin DNA will be introduced into a tumour cell by transfection with polyethylenimine (PEI). The PEI is a stable cationic polymer. PEI will condense DNA into positively charged particles, and consequently, the (PEI)-DNA complex will be endocytosed by the cells. The efficiency of transfection will be monitored using GFP expression by flow cytometry. This will produce green light. Also, we assessed cytotoxic effects of transfection reagents. We can conclude that the transfection is more efficient with a 1:1 PEI:DNA ratio, except in the HMLE Twist cell line. Furtermore, it will be better to use flow cytometry for both the transfection efficiency and cytotoxic effects of apoptin in the CSCs.

Izvorni jezik
Engleski

Znanstvena područja
Biologija

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