Determination of Cytotoxic effect and Expression analyses of Apoptotic and Autophagic related genes in Thymoquinone-treated Colon Cancer Cells

Year 2020, , 189 - 196, 01.02.2020

Esra Tokay

https://doi.org/10.16984/saufenbilder.585012

Cited By: 1

Abstract

Thymoquinone
(Tq; 2-isopropyl-5-methyl-1,4 benzoquinone) is the main ingredient present in
the volatile oil of Nigella sativa
(black seed). Recently, it has been reported that, Tq was inhibited cell
proliferation of many cancer cell lines including ovarian, lung, breast, osteosarcoma,
pancreatic, fibrosarcoma, lung cancer, squamous cell carcinoma and myeloblastic
leukemias. In this study, the anticancer effect
of Tq was investigated in human Colon Cancer Cells (HT-29) with MTT assay.
Positive control was achieved with 5-fluorouracil (5-FU). Also, the invasion
capability of HT-29 cellls was determined in presence of Tq and 5-FU with
scratch analyses. In addition, the autophagic and apoptotic effect of Tq were
investigated by qRT-PCR method to find out the pathway of anti-proliferative
effect in Tq-treated HT-29 cells. As a result, Tq was inhibited the cell
proliferation of HT-29 cells in time dependent manner, with 118  µM for 24 h and 84 µM for 48 h, respectively. A
concentration of 150 µM of Tq was able to significantly reduced the
invasiveness of HT-29 cells about  ̴8
fold compared with not treated cells at 6h. ATG-12, ATG-7 and LC3-II were
significantly downregulated in presence of Tq.  Conversaly, the expression of all of the
autophagy genes were downregulated in presence of 5-FU. Moreover, pro-apoptotic
gene Bax was significiantly upregulated nearly 15 fold whereas the expressions
of Bcl-2 and Bcl-XL (pro-survival genes) were decreased in presence of Tq. On
the other hand, in 5-FU treated HT-29 cells, the expression of  Bax, Bcl-2 and Bcl-XL genes and
autophagy related genes were significantly downregulated. 

Keywords

Tq, autophagy, apoptosis, HT-29

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