Frequency of Mutations in BRAFV600E, RASAl1 and RASA1 Genes in Tumor and Peripheral Blood Samples of the Patients with Ovarian Cancer Using Real-Time PCR Technique

Abstract

Ovarian cancer is the fourth most prevalent cancer and the leading cause of death because of gynecological cancers, whose epithelial type is the sixth most common cancer among women. Although the etiologic causes of ovarian cancer are not recognized well, genetic precursors like changes in the KRAS / BRAF pathway causing mutations in the type I biological evolution pathway of ovarian cancer especially in somatic cells are worth examining and significant. The study examined RASA₁ and RASA1₁ genes from the KRAS gene family that code protein and are the main components of the MAPK / RAS tyrosine-kinase pathway. Common point mutations in these genes disrupt GTPase function ending in the loss of protein inactivation ability.

Methods: This case-control study examined 25 ovarian tumor samples, 25 blood samples of ovarian cancer patients that had undergone surgery, and 50 blood samples from healthy women, five samples from normal ovarian tissue as controls. DNA extraction was then carried out for all the samples and finally, the mutation frequency of RASA1₁, RASA1, and BRAFV600E genes was examined using Quantitative Real-time PCR.

Results: No mutations were seen in RASAl₁ and RASA1 in tumor samples and their blood samples after the analysis of the findings obtained from the real-time PCR technique by relative method on tumor tissue and blood samples of the patient group with ovarian cancer, normal tissue samples as the control, and blood of healthy female group in terms of ovarian cancer. No mutations were found in BRAFV600E in tumoral tissue, yet two patients had this mutation (8%) in the blood sample of this group. No mutations in these three genes were found in the blood sample of healthy people.

Conclusion: Despite the presence of the mutations in other tumors, they do not have the potential to be introduced as research objectives in biomarker detection studies for early detection of metastasis in patients with ovarian cancer given the lack of mutations in RASAl₁, RASA1, and BRAFV600E. Nonetheless, considering the BRAFV600E gene and the mutations in the blood samples of two patients and its 8% prevalence, more examination is recommended in this case as a less invasive and early method in early diagnosis of cancer.