TY  - JOUR
T1  - Determination of Cytotoxic effect and Expression analyses of Apoptotic and Autophagic related genes in Thymoquinone-treated Colon Cancer Cells
AU  - Tokay, Esra
PY  - 2020
DA  - February
Y2  - 2019
DO  - 10.16984/saufenbilder.585012
JF  - Sakarya University Journal of Science
JO  - SAUJS
PB  - Sakarya University
WT  - DergiPark
SN  - 2147-835X
SP  - 189
EP  - 196
VL  - 24
IS  - 1
LA  - en
AB  - Thymoquinone(Tq; 2-isopropyl-5-methyl-1,4 benzoquinone) is the main ingredient present inthe volatile oil of Nigella sativa(black seed). Recently, it has been reported that, Tq was inhibited cellproliferation of many cancer cell lines including ovarian, lung, breast, osteosarcoma,pancreatic, fibrosarcoma, lung cancer, squamous cell carcinoma and myeloblasticleukemias. In this study, the anticancer effectof Tq was investigated in human Colon Cancer Cells (HT-29) with MTT assay.Positive control was achieved with 5-fluorouracil (5-FU). Also, the invasioncapability of HT-29 cellls was determined in presence of Tq and 5-FU withscratch analyses. In addition, the autophagic and apoptotic effect of Tq wereinvestigated by qRT-PCR method to find out the pathway of anti-proliferativeeffect in Tq-treated HT-29 cells. As a result, Tq was inhibited the cellproliferation of HT-29 cells in time dependent manner, with 118 µM for 24 h and 84 µM for 48 h, respectively. Aconcentration of 150 µM of Tq was able to significantly reduced theinvasiveness of HT-29 cells about ̴8fold compared with not treated cells at 6h. ATG-12, ATG-7 and LC3-II weresignificantly downregulated in presence of Tq. Conversaly, the expression of all of theautophagy genes were downregulated in presence of 5-FU. Moreover, pro-apoptoticgene Bax was significiantly upregulated nearly 15 fold whereas the expressionsof Bcl-2 and Bcl-XL (pro-survival genes) were decreased in presence of Tq. Onthe other hand, in 5-FU treated HT-29 cells, the expression of Bax, Bcl-2 and Bcl-XL genes andautophagy related genes were significantly downregulated.
KW  - Tq
KW  - autophagy
KW  - apoptosis
KW  - HT-29
CR  - [1] A. Bhandari, M. Woodhouse, and S. Gupta, “Colorectal cancer is a leading cause of cancer incidence and mortality among adults younger than 50 years in the USA: a SEER-based analysis with comparison to other young”, J Investig Med. vol.65, no.2:, pp. 311–315, 2017.
CR  - [2] T.J Syed Ahmad Tajudin., H. Fatimah, I. Wan Iryani and A. Abdul Manaf,  “Combinatorial Cytotoxic Effects of Gelam Honey and 5-Fluorouracil against Human Adenocarcinoma Colon Cancer HT-29 Cells In Vitro’. International Journal of Cell Biology”, vol. 2019, pp.10, 2019.
CR  - [3] A.E. Ashour, A.R. Abd-Allah, H.M. Korashy, S.M. Attia , Alzahrani A.Z., Saquib Q., Bakheet S.A., Abdel-Hamied H.E., Jamal S., Rishi A.K., “Thymoquinone suppression of the human hepatocellular carcinoma cell growth involves inhibition of IL-8 expression, elevated levels of TRAIL receptors, oxidative stress and apoptosis”. Mol Cell Biochem. vol. 389 no.1-2,pp. 85-98, 2014
CR  - [4] Raghunandhakumar S., Paramasivam A., Senthilraja S., Naveenkumar C., Asokkumar S., Binuclara J., Jagan S., Anandakumar P., Devaki T.,”Thymoquinone inhibits cell proliferation through regulation of G1/S phase cell cycle transition in N-nitrosodiethylamine-induced experimental rat hepatocellular carcinoma”. Toxicol Lett., vol.223, no.1,pp.60-72,2013.
CR  - [5] Attoub S., Sperandio O., Raza H., Arafat K., Al-Salam S., Al Sultan M.A., Al Safi M., Takahashi T., Adem A., “Thymoquinone as an anticancer agent: evidence from inhibition of cancer cells viability and invasion in vitro and tumor growth in vivo”. Fundam Clin Pharmacol., vol. 27, no. 5, pp. 557-69,2013.
CR  - [6] Shahin Y.R., Elguindy N.M., Abdel Bary A., Balbaa M., “The protective mechanism of Nigella sativa against diethylnitrosamine-induced hepatocellular carcinoma through its antioxidant effect and EGFR/ERK1/2 signaling”. Environ Toxicol. doi: 10.1002/tox.22574. [Epub ahead of print],2018.
CR  - [7] Mohamed M S.A., Abdulaziz M.A., Salim S A.R., Abdulaziz A. Al-Y., Othman A. Al-S., Mohamed M.H., Mahmoud N.N., “Thymoquinone attenuates diethylnitrosamine induction of hepatic carcinogenesis through antioxidant signaling”. Oxid Med Cell Longev. Vol.  3, no. 2 , pp. 254–261., 2010.
CR  - [8] El-Baba C., Mahadevan V., Fahlbusch F.B., Mohan S.S., Rau T.T., Gali-Muhtasib H., Schneider-Stock R., “Thymoquinone-induced conformational changes of PAK1 interrupt prosurvival MEK-ERK signaling in colorectal cancer”. Mol Cancer., vol. 13, pp. 201, 2014.
CR  - [9] Fischer T.D., Wang J.H., Vlada A., Kim J.S., Behrns K.E.,”Role of autophagy in differential sensitivity of hepatocarcinoma cells to sorafenib”. World J Hepatol. Vol. 6, no. 10, pp. 752-8,2014.
CR  - [10] Juan W., Qi W., Juan-Juan L., Chuang C., Si S., Chang-Hua W., Sheng-Rong S., “Autophagy mediates free fatty acid effects on MDA-MB-231 cell proliferation, migration and invasion”. Oncol Lett. Vol. 14, no. 4, pp. 4715–4721, 2017.
CR  - [11] Liu J., Chen Z., Guo J., Wang L., Liu X., “Ambra1 induces autophagy and desensitizes human prostate cancer cells to cisplatin”. Biosci Rep. doi: 10.1042/BSR20170770. [Epub ahead of print], 2017.
CR  - [12] Wan B., Dai L., Wang L., Zhang Y., Huang H., Qian G., Yu T., “Knockdown of BRCA2 enhances cisplatin and cisplatin-induced autophagy in ovarian cancer cells”. Endocr Relat Cancer. Vol. 25, no. 1, pp. 69-82, 2018.
CR  - [13] Tokay E. and Kockar F. “Identification of intracellular pathways through which TGF-β1 upregulates URG-4/URGCP gene expression in hepatoma cells”, Life Sci., vol. 144, pp. 121-8, 2016.
CR  - [14] Schneider C. A.; Rasband W. S. and Eliceiri, K. W., “NIH Image to ImageJ: 25 years of image analysis”, Nature methods, vol. 9, no. 7, pp. 671-675, 2012.
CR  - [15] El-Najjar N., Chatila M., Moukadem H., Vuorela H., Ocker M., Gandesiri M., Schneider-Stock R., Gali-Muhtasib H., “Reactive oxygen species mediate thymoquinone-induced apoptosis and activate ERK and JNK signaling”, Apoptosis, vol. 15, no. 2, pp. 183-95, (2010).
CR  - [16] Chen C., Lu L., Yan S., Yi H., Yao H., Wu D., He G., Tao X., Deng X. Autophagy and doxorubicin resistance in cancer. Anticancer Drugs, vol. 29, no. 1, pp. 1-9, 2018.
UR  - https://doi.org/10.16984/saufenbilder.585012
L1  - https://dergipark.org.tr/en/download/article-file/967514
ER  -