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Evaluation of the expression of several genes involved in cancer in several different cancer cell lines by RT-PCR

Document Type : Article

Authors

1 Sc. of Biochemistry, Department of Biology, Payamnoor University, Tehran, Iran

2 Professor of Biochemistry Department, Payamnoor University

3 Ph.D. of Biochemistry, University of Georgia

4 M.Sc. of Molecular Biology, Emam Hosein University

5 M.Sc. Student, Genetic Department, Tarbiat Modares University

Abstract

  Abstract
The altered or mutated forms of genes known as proto-oncogenes are responsible for promoting cell growth and uncontrolled cell proliferation. An accumulation of many mutations in different and specific genes,over time is required to cause cancer. The pattern of gene expression, also called molecular signature is unique to a particular class of tumor or tumor cell. This paper describes the latest technique for monitoring the expression of a panel of cancer-specific genes. The PCR technique combines the quantitative performance of SYBR® Green-based real-time PCR is widely used for gene profiling. This technique is cost-effective, easy-to-use, and focuses only on the genes that you desire. In this study the expression of our target genes were quantitatively determined in five human cancer cell lines. We selected gene β-actin as our reference gene. Cells were lysed and the mRNAs were extracted using the RNA Purification Kit and cleaned up with Qiagen RNeasy spin columns. The first-strand cDNA was synthesized according to the High Capacity cDNA Reverse Transcription Kit protocol. RT-PCR were performed with Gene Expression Assays in an AB step one plus Sequence Detection System. Briefly the expression of p53 was high in both breast cancer cell lines, MCF7, T47-D and lung cancer cells, A549. Src expression was higher in prostate cell line, PC3 and lung cancer cells, A549. Meanwhile SKOV3 (ovarian cancer cell line) showed high expression of her-2 gene.  The results clearly show that the expression pattern of this panel of genes was unique to almost every cell line examined.
 

Keywords

References
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Experimental animal Biology
Volume 3, Issue 3 - Serial Number 3
May 2015
Pages 1-12
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