Gene Expression Study on Similarity Effect of Thymoquinone on Epithelial Breast Cancer and Colorectal Cancer Cell Lines
Issue:
Volume 3, Issue 2-2, March 2015
Pages:
1-6
Received:
9 December 2014
Accepted:
17 December 2014
Published:
27 December 2014
Abstract: Adenocarcinoma is known as a common type of cancer which includes 85% of breast carcinoma and 95% of colorectal carcinoma. Up until now, the cytotoxic effect of Thymoquinone on different types of tumor cells has been reported. It was hypothesized that Thymoquinone has similar effect on cancer arising from epithelial cells through gene expression analysis in MCF7 breast cancer and HT-29 colon cancer cell lines. The quantity and quality of RNA samples were identified using RNeasyPlus Mini kit and RNA 6000 Nano LabChip kit, respectively. The purified RNA samples of MCF7 cells were used in two-color 8×60K cDNA array platform with SurePrint Agilent technology. The hybridized cRNA/cDNA probes were identified due to labeling with red and green cyanine dyes using GEX hybridization buffer HI-RPM. LOWESS normalization reduced the dye bias on the array slide using feature extraction software. Gene ontology analysis was done after performing different steps of filtering to reduce not satisfied genes using Gene Spring software. Two-step RT-qPCR assay using Taq Man fast advanced master mix analyzed the most up and down regulated genes in MCF7 compared to HT-29 cancer cell lines. The 10 different cancer cell lines in a form of universal reference RNA was used as standard data set comparison and as a positive control RNA in cDNA array and RT-qPCR assay, respectively. The T-test statistical analysis of independent samples showed that there is no significant difference between two types of cell lines due to Thymoquinone treatment with p-value 0.844. Among the selected genes; CARD16, UGT1A8, SLC7A11, IFIT1, IF16 and IFIT3 were expressed significantly (0.009, 0.0001, 0.037, 0.098, 0.0001 and 0.033, respectively) in breast cancer compare to colon cancer cells. The findings indicated similar effect of Thymoquinone on cancer arising from epithelial cells.
Abstract: Adenocarcinoma is known as a common type of cancer which includes 85% of breast carcinoma and 95% of colorectal carcinoma. Up until now, the cytotoxic effect of Thymoquinone on different types of tumor cells has been reported. It was hypothesized that Thymoquinone has similar effect on cancer arising from epithelial cells through gene expression an...
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Cytotoxic, Cytostatic and Anti-Estrogenic Effect of Thymoquinone on Estrogen Receptor-Positive Breast Cancer MCF7 Cell Line
Issue:
Volume 3, Issue 2-2, March 2015
Pages:
7-14
Received:
10 December 2014
Accepted:
13 December 2014
Published:
27 December 2014
Abstract: About 80% of breast cancers are estrogen-receptor positive. The research carried out herein focused on the effect of Thymoquinone which is an active compound of Nigella sativa seed on estrogen-receptor positive breast cancer MCF7 cell line. The percentage of apoptotic cells was found using Annexin V-FITC apoptosis detection kit. CycleTEST PLUS DNA Reagent was used to distinguish distribution of treated cells between different cell cycle phases. DNA microarray identified the regulated genes, level of expressed genes, gene ontology and pathway networks. Significant arrest of treated cells at G1 phase suggested cytostatic effect of Thymoquinone 100 µM after 24 hours at p-value < 0.05 which was similar to anti-estrogenic compounds such as Tamoxifen. Cytotoxic effect of Thymoquinone 100 µM was found through highly significant accumulation of cells at sub-G1 phase after 72 hours at p-value < 0.0001. CYP1A1, CYP1B1, NQO1 and UGT1A8 genes were down regulated after 24 hours treatment with Thymoquinone 50 µM concentration which suggested reduction of catechol estrogens and rising in metoxy forms of estradiol and estrone. Reduction of ER would be predictable due to the down-regulation of CYP1B1 and UGT1A8 genes which reduced affinity of trans-tamoxifen-o-glucuronide to ER. The study proposed the benefits of using Thymoquinone to accelerate Tamoxifen effects in treating breast cancer and reducing its side effects.
Abstract: About 80% of breast cancers are estrogen-receptor positive. The research carried out herein focused on the effect of Thymoquinone which is an active compound of Nigella sativa seed on estrogen-receptor positive breast cancer MCF7 cell line. The percentage of apoptotic cells was found using Annexin V-FITC apoptosis detection kit. CycleTEST PLUS DNA ...
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