Expression analysis of mir-21 and mir-221 in cancerous tissues from Iranian patients with gastric cancer

Early detection is a key to survival for gastric cancer. Molecular markers such as miRNA [microRNA] can have great importance in the early diagnosis of gastric cancer. Expression of miR-21 and miR-221 are deregulated in many types of human cancers. This study aimed to investigate the differences in miRNA expression patterns within the Iranian population. Total RNA was extracted from gastric cancer tissues and adjacent non-cancerous tissues from 32 patients. Expression levels of miR-21 and miR-221 were detected by Real time RT-PCR using a specific primer, with 5s rRNA as the internal reference gene. Our data showed that the expression levels of miR-21 and miR-221 in gastric cancer samples were significantly higher than in paired non-cancerous samples [P < 0.05]. The receiver operating characteristic [ROC] analyses yielded the area under the curve [AUC] values of 80.30 for miR-21 and 93.30 for miR-221, and combined ROC analysis revealed the highest AUC value of 96.90 in discriminating GC patients from healthy controls. It seems that miR-21 and miR-221 expression pattern in Iranian patients with gastric cancer are similar to any other population. Considering the increased expression level of two miRNAs in cancerous tissue compared to normal tissue as well as the area under ROC curve, miR-21 and miR-221 can be used for early detection of gastric cancer

Expression of two basic mRNA biomarkers in peripheral blood of patients with non-small cell lung cancer detected by real-time RT-PCR,individually and simultaneously

Although extensive research has been conducted on lung cancer markers, a singular clinically applicable marker has not been found yet. The objective of this study was to evaluate the sensitivity and the specificity of carcinoembryonic antigen [CEA] mRNA and lung-specific X protein [LUNX] mRNA biomarkers in peripheral blood to detect lung cancer individually and simultaneously.Thirty patients affected by lung cancer and 30 healthy individuals were studied in this research. Three vials of cDNA were made from each sample after taking peripheral blood samples and extracting total RNA. Each sample was examined by the real-time RTPCR technique. The result from each vial was then compared with the sensitivity of overall marker. The CEA mRNA was positive in 24 out of 30 lung cancer patients. Hence, its sensitivity was determined at 80%,differing significantly from that observed in healthy individuals, where 11 positive cases were seen. The overall sensitivity of this marker was significantly associated with positivity in vials 2 and 3 but not in vial 1 The LUNX mRNA was positive in 21 out of 30 patients, indicating 70% sensitivity. This finding significantly differed from that in healthy individuals. The overall sensitivity of this marker was significantly associated with positivity in vials 1 and 3, but not in vial 2. In 93.3% of the patients, at least one positive marker was observed.The mentioned mRNA could be suggested as sensitive and specific markers in peripheral blood for primary diagnosis of lung cancer.

Simultaneous analysis of multidrug resistance 1[MDR1] C3435T, G2677T/A, and C1236T genotypes in Hamadan city population, west of Iran

One of the limitations in the treatment of common diseases such as cancer chemotherapy is development of multidrug resistance [MDR]. Polymorphisms could alter the expression level of MDR1 gene, which plays an important role in MDR. In this research, the frequency of C3435T, C1236T, and G2677T/A polymorphisms of MDR1 gene was investigated in a large group of population from Hamadan city to provide a sample data resource.Peripheral blood [2 ml] was taken, and DNA extraction was carried out. Multiplexed mutagenically separated PCR, which was followed by polyacrylamide gel electrophoresis and silver staining, was applied to detect the mentioned polymorphisms in 935 individuals. Sequencing performed for confirmation of gel electrophoresis resulted in 10 random cases. In total, alleles and genotypes of 933 persons [776 women and 157 men] were determined. The most frequent alleles of the polymorphisms were: 3435T, C1236, and G2677. The most frequent genotypes were: 3435C/T, 1236C/T, and 2677G/A, and their concurrent presence was also found as the most frequent simultaneous genotypes. There was not any meaningful difference among the prevalence of these genotypes in groups of men and women. Our results were close to those of other studies performed in Iran and compared to the other ethnic groups, which showed more similarity to Asian peoples than Europeans. As an aspect of personalized medicine, it could be used by chemotherapists to improve the routine methods of cancer treatment.

Effect of myomectomy on endometrial glutathione peroxidase 3 [GPx3] and glycodelin mRNA expression at the time of the implantation window

In fertile women, glycodelin and glutathione peroxidase 3 [GPx3] genes expression rises during the luteal phase, with a peak occurring during the implantation window. The expression of these genes decreases in women with myomas. To determine whether myomectomy would reverse glycodelin and GPx3 expression, we evaluated the transcript levels of these genes in the endometrium of patients before and after myomectomy. Expression of glycodelin and GPx3 genes were examined prospectively during the midluteal phase in the endometrium obtained from infertile women with myoma [n = 12] before and three months after myomectomy. Endometrial expression of these genes was evaluated using quantitative real-time RT-PCR. Endometrial glycodelin mRNA expression levels [normalized to 18S rRNA expression] were increased significantly in endometrium of patients after myomectomy [P = 0.02]. GPx3 mRNA expression was increased insignificantly after myomectomy [P = 0.43]. The results showed that myomectomy increased endometrial glycodelin [significantly] and GPx3 [not significantly] gene expression after 3 months. Study at different times and detecting expression of these genes can reveal more details

Improved real-time RT-PCR assays of two colorectal cancer peripheral blood mRNA biomarkers: a pilot study

Background: efficient screening for detection of colorectal cancer [CRC] at earlier stages reduces its mortality. The purpose of this study was to investigate expression of carcinoembryonic antigen [CEA] and human telomerase reverse transcriptase [hTERT] mRNA in peripheral blood of CRC patients and to present strategies for early detection screen test

Methods: twenty seven patients in non-metastatic stage and 27 healthy individuals were studied. Expression of CEA, hTERT mRNA and 18srRNA [18s subunit of ribosomal RNA, as reference gene] were determined based on real-time RT-PCR on 3 [micro]g of total RNA from blood in 3 separate vials [1 [micro]g per vial]

Results: positive expression rate of CEA mRNA [78%] and hTERT mRNA [81%] were higher in patient group [P<0.001]. These rates were meaningfully higher than the results of individual vials containing only 1 [micro]g of total RNA. Difference between Ct values of markers with 18srRNA [[DELTA]Ct] was higher in healthy group than patient one. Therefore, a [DELTA]Ct cut-off value was determined for distinguishing between true- and false-positive results. Concurrent expression of both markers was found in 67% of the patients, which was higher than healthy cases [11%]. Combination of concurrent marker expression with cut-off point strategy increased specificity to 100%

Conclusion: these results showed that concurrent evaluation of marker expression and performing the test on 3 [micro]g of samples in 3 separate vials may increase specificity and sensitivity of real-time RT-PCR for early detection of non-metastatic CRC. However, more investigations with larger numbers of samples are needed to verify these results. Iran. Biomed. J. 17 [1]: 15-21, 2013