Influence of exogenous overexpression of p21WAF1/Cip1 gene and various chemotherapeutics on proliferation of colon cancer cells

Stulić, Maja (2010) Influence of exogenous overexpression of p21WAF1/Cip1 gene and various chemotherapeutics on proliferation of colon cancer cells. Diploma thesis, Faculty of Science > Department of Biology.

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Abstract

p21WAF1/Cip1 protein is primarily defined as a cyclin dependent kinase inhibitor which induces cell cycle arrest under stress conditions (DNA damage, oxidative stress, anticancer agents...). Recent studies point out an important role of p21 protein in regulation of DNA replication as well as very complex part in induction or inhibition of cell death, which depends on tretment type, cell type and genetic context. Since p21Waf1/Cip1 revealed equivocal and somewhat controversial role, it is a major issue to elucidate mechanism of it`s action after DNA damage caused by common chemotherapeutics in order to establish the influence of p21 to cell chemosensitivity , which coluld give rise to more targeted tumor therapy. In our expreiments we investigated the effects of five common chemotherapeutics and overexpression of exogenous p21WAF1/Cip1 on the growth of colon cancer cell lines HCT116, SW480, SW620, CaCo-2. Enhanced expression of protein p21WAF1/Cip1 was accomplished by infection with replication-defective adenoviral recombinant vector Ad5CMVp21, whereas Ad5CMVβ-gal was used as a control vector. In order to evaluate the effect of exogenous protein p21WAF1/Cip1 on the sensitivity to chemotheraputics, we treated cells with chemotherapeutics camptothecin, doxorubcin and cisplatin at different concetration range after infection wtih adenoviral vectors. Cells’ viability was monitored by MTT assay, and the expression of p21 protein by Western blot method. All of the tested chemotherapeutics inhibited growth of the tumor cells whereby 5-fluorouracil revealed least and camptothecin strongest effect. We detected significant growth inhibition of HCT116 and SW620 cells five days after infection with Ad5CMV-p21 at MOI 60. Furthermore, p21WAF1/Cip1 gene overexpression in HCT116 and SW620 cells distinctly enhanced cisplatin and camptotecin influence, whereas it had faint effect on doxorubicin influence. We detected similar but weaker effect of p21 on CaCo-2 cells. On the other hand, even though SW480 appeared to be the most sensitive to chemotherapeutics, their effect wasn`t enhanced when combined with p21WAF1/Cip1 overexpression in this cell line.

Item Type: Thesis (Diploma thesis)
Keywords: gene p21WAF1/Cip1, chemotherapy, tumor, cell cycle, adenoviral vectors
Supervisor: Kralj, Marijeta
Co-supervisor: Matulić, Maja
Date: 2010
Number of Pages: 54
Subjects: NATURAL SCIENCES > Biology
Divisions: Faculty of Science > Department of Biology
Depositing User: Silvana Šehić
Date Deposited: 04 Sep 2014 10:52
Last Modified: 04 Sep 2014 10:52
URI: http://digre.pmf.unizg.hr/id/eprint/2625

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