The Relevance of Common K-RAS Gene Mutations and K-RAS mRNA Expression with Clinicopathological Findings and Survival in Breast Cancer.

INTRODUCTION: Studies have shown the role of mutation and gene expression of K-RAS in luminal breast cancer. In the current study, the status of common K-RAS mutations and mRNA expression in breast cancer were investigated. The aim of this research was determining the relationship of these molecular finding with clinicopathological features and 5 year overall survival. MATERIAL AND METHODS: In this case control study, we examined tumor tissue obtained from patients who had breast surgery which their paraffin tissue samples were available in the pathology department. Samples who had codon 12 and 13 mutations in exon 2 of K-RAS gene were considered as case group and tumor tissues without these mutations were considered as control group. The expression of K-RAS mRNA was explored by real-time polymerase chain reaction (RT-PCR) and the results were reviewed with clinicopathological features and survival of patients. RESULTS: The results of the present study showed that 14% and 10% of patients had K-RAS mutations in codons 12 and 13, respectively. There was a significant relationship between K-RAS mutations with T staging and PR positivity in tumors. Five years overall survival was 8% in case group compare to control group who had 69% 5y OS. Furthermore, K-RAS mRNA expression had a significant relationship with T and N staging and 5 year survival. In conclusion, it seems that two molecular markers of mutation and K-RAS gene expression may be used simultaneously to estimate the prognosis of breast cancer. CONCLUSION: It seems that two molecular markers of mutation and K-RAS gene expression may be used simultaneously to estimate the prognosis of breast cancer.

A Comprehensive Comparison of Rapid RNA Extraction Methods for Detection of SARS-CoV-2 as the Infectious Agent of the Upper Respiratory Tract using Direct RT-LAMP Assay.

Background: The current COVID-19 pandemic has highlighted the need for faster and more cost-effective diagnostic methods. The RNA extraction step in current diagnostic methods, such as real-time qPCR, increases the cost and time required for testing. Reverse-transcription loop-mediated isothermal amplification (RT-LAMP) is a promising technique for developing diagnostic tests with desired sensitivity and specificity without the need for RNA extraction. Materials and Methods: An RT-LAMP assay was developed to detect SARS-CoV-2 with a sensitivity of 0.5 copies of positive control plasmid per microliter in 40 min. Several rapid RNA extraction protocols were evaluated using different reagents, including bovine serum albumin, Triton X-100, Tween 20, proteinase K, guanidine hydrochloride, guanidinium isothiocyanate (GITC), and thermal treatment. Finally, the sensitivity and specificity of the developed direct RT-LAMP were determined using 150 upper respiratory tract samples. Results: Method 10 was selected as the most efficient protocol for the RNA extraction step. The sensitivity and specificity of the developed direct RT-LAMP assay with clinical samples were estimated at 98.4% and 88.8%, respectively. Conclusion: These results suggest that the combination of GITC and Triton X-100 detergent is a highly efficient method for RNA extraction and direct RT-LAMP detection of SARS-CoV-2 in clinical samples, providing a valuable tool for the rapid and cost-effective diagnosis of COVID-19.

Correlation of miR-31 and miR-373 expression with KRAS mutations and its impact on prognosis in colorectal cancer.

INTRODUCTION: Colorectal cancers (CRC) are among the most common cancers. There are different modalities for treatment including chemotherapy, surgery, and radiotherapy. There are some mutations in cancers which can assist in the treatment and better prognosis of patients. In this study, two molecular markers (miR-31 and miR-373) were involved in the pathogenesis of CRC and their association with histopathological features was investigated. As well, the prognostic value of these molecular markers was investigated in CRC patients with or without common KRAS mutations. METHODS: Paraffin blocks of tissue samples from 150 patients who underwent colon surgery between 2018 and 2020 were prepared by the Pathology Department of Imam Hossein Hospital (Tehran, Iran). After DNA and RNA isolation, gene expression of miR-31 and miR-373 was determined using probe-based quantitative real-time polymerase chain reaction (qRT-PCR). Mutations of KRAS were surveyed using conventional PCR and agarose gel electrophoresis. RESULTS: The mean age of the patients was 57.2 ± 13.4 years. KRAS codon 12 and 13 mutations were positive in 31 (20.6%) and 22 (14.6%) cases, respectively. The results showed that KRAS common mutations occurred in 32.6% of Iranian CRC patients. The expression levels of miR-31 and miR-373 increased in CRC patients with KRAS mutations in comparison with patients without these mutations. CONCLUSION: Considering the role of miR-31 and miR-373 in CRC tumor progression, it seems that the CRC patients bearing KRAS mutations have a poorer prognosis respective to patients without KRAS mutations.

Long non-coding RNA panel as a molecular biomarker in glioma.

BACKGROUND: Glioma is one of the most malignant brain tumors, accounting for about half of the gliomas that occur in central nervous system (CNS), originates from the glial tissue of the brain. The aim of the present study was to determine the expression levels of 5 lncRNAs (MDC1-AS1, HOXA11-AS, MALAT1, CASC2, ADAMTS9-AS2) in patients with high-grade glioma in comparison with low grade glioma. METHODS: This was a retrospective study which determined molecular biomarker on pathologic glioma samples. We examined 100 patients' pathologic block which consisted of 50 pathology samples of high-grade glioma (case group) and control group consisted of 50 pathology samples of low-grade glioma. This research was performed using real time polymerase chain reaction (PCR) technique. RESULTS: The results showed that the expression of ADAMTS9-AS2 and HOXA11-AS genes significantly increased with increasing tumor grade. Also the expression of CASC2 gene significantly decreased with increasing tumor grade. CONCLUSIONS: It was concluded that ADAMTS9-AS2 and HOXA11-AS and CASC2 are promising lncRNA markers in prognosis of glioma.