بررسي بيان ژن CAMK2N1 در نئوپلاسم تيروئيد در جمعيت اصفهان

Thyroid neoplasm is the most common malignancy of the endocrine system in humans, and the most common type is papillary thyroid cancer (PTC). Due to the rapid increase in this neoplasm, it is predicted that this disease will become the most common type of cancer worldwide. The genetic mechanism of PTC is not fully understood. Therefore, finding key genes in this disease can be more important. Analyzing large biological data, including microarray data, can be very important to find genes with different expression in diseased and healthy samples. In this study, 12 GSEs were loaded from the NCBI GEO database. Then data normalization was done by Log2 Transformation and Quantile Normalization. Based on this, by analyzing the microarray data and using the Limma package, we found differentially expressed genes (DEG). Then, by dividing among the common genes related to these analyses, significant key genes (p.value < 0.05) with different expression between PTC tissues and adjacent normal tissue were obtained. In laboratory tests, the expression level of CAMK2N1 gene in 20 samples, including 10 PTC tumor samples and 10 healthy tissue samples adjacent to the tumor, was checked by RT-qPCR technique. In order to design the primers, Beacon Designer 8.1 and Oligo 7 software were used. By determining the crossing regions of the exons, the design of the intersection of the primers was done. Tm temperature was also optimized. The RT-qPCR step in this study was performed using the Rotor-Gene Q device and the help of SYBR Green fluorescent dye. The results of this eva‎luation were statistically analyzed using REST 2009 software. Based on the obtained results, a significant increase in the expression of CAMK2N1 gene by 3.596 times was observed in PTC samples compared to healthy tissue (p.value=0.012). The level of changes in CAMK2N1 gene expression in relation to PTC varieties, tumor size, age and history of hypothyroidism in PTC patients was not statistically significant. With this significant increase in expression 5.657 times was observed in PTC samples with metastasis (N1) compared to PTC samples without metastasis (N0) (p.value=0.001).

4

33946