Human Gene expression profiling using cDNA microarray in estrogen receptor positive breast cancer MCF7 cell line treated with Thymoquinone

Tumor classification, identification of new diagnostic markers, identification of cellular pathways that are affected by malignant transformation can be revealed through gene expression profiling. Gene expression profile can provide a molecular signature of specific tumor, in diseased cells and patient samples. It also provides information on the pathology of disease, resistance to cure and response to cellular micro environments. Gene expression analysis based on the array platform is a powerful tool to profile expression of genes and to generate information on therapeutic effects of new anti-cancer compounds. Micro array technique has also been introduced as a gene profiling method to enable the detection of molecular relations between genes through specific pathways. In this study, micro array based diagnostic method was used to differentiate between two separate profiles of estrogen receptor positive breast cancer cells, MCF7 cell line without treatment and after treatment with Thymoquinone. In previous researches, the toxicity influence of Thymoquinone revealed its potential anti-cancer effect on breast cancer. In the current research, after culturing breast cancer MCF-7 cell line, sufficient amount of pureed RNA was extracted from the cells using RNeasy Plus Mini kit and Agilent RNA 6000 Nano Lab Chip kit. The Sure Print Agilent technology was used by performing two-color cDNA micro array 8 well×60,000 base platform. The cDNA strands were generated using reverse transcription promoter primer. cDNA converted to cRNA using T7 RNA polymerase. cRNA samples were labelled with Cy-3-CTP produced red color and human universal reference RNA was labelled with Cy-5-CTP as a positive control RNA sample produced green color. Identification of two hybridized RNA samples to cDNA probes on the array platform was done using GEX hybridization buffer HI-RPM based on the colors. Filtering, normalization and statistical steps were done automatically using Agilent technology which showed the altered genes. The findings could be differentiated with the EST profile of breast cancer and revealed the anti-cancer effect of compound. These findings emphasized that detailed analysis of entire genome of breast cancers before treatment and after treatment by micro array would offer potential application in personalized medicine.

کلیدواژه ها:

gene expression profiling ، personalized medicine ، cDNA microarray ، Agilent technology ، breast cancer ، Thymoquinone