ABSTRACT
Background: The cyclin E2 [CYCE2] is an important regulator in the progression and development of NSCLC, and its ectopic expression promoted the proliferation, invasion, and migration in several tumors, including Non-Small Cell Lung Cancer [NSCLC]. However, the upregulation of CYCE2 in NSCLC cells suggested that it has a key role in tumorigenicity. In addition, the RAS family proteins as oncoproteins were activated in many major tumor types and its suitability as the therapeutic target in NSCLC was proposed. Considering the crucial role of microRNAs, it was hypothesized that altered expression of hsa-miR-30d-5p and hsa-let-7b might provide a reliable diagnostic tumor marker for diagnosis of NSCLC
Method: Real-time RT-PCR approach could evaluate the expression alteration of hsa-miR-30d-5p and hsa-let-7b and it was related to the surgically resected tissue of 24 lung cancer patients and 10 non-cancerous patients. The miRNAs expression was associated with clinicopathological features of the patients
Results: Hsa-miR-30d showed a significant downregulation [p=0.0382] in resected tissue of NSCLC patients compared with control group. Its expression level could differentiate different stages of malignancies from each other. The ROC curve analysis gave it an AUC=0.73 [p=0.037] which was a good score as a reliable biomarker. In contrast, hsa-let-7b was significantly overexpressed in tumor samples [p=0.03]. Interestingly, our findings revealed a significant association of hsa-let-7b in adenocarcinoma tumors, compared to Squamous Cell Carcinomas [SCC] [p<0.05]. Also, analysis of ROC curve of hsa-let-7b [AUC=0.74, p-value=0.042] suggests that it could be as a suitable biomarker for NSCLC
Conclusion: Together, these results suggest a possible tumor suppressor role for hsa-miR-30d in lung tumor progression and initiation. Moreover, upregulation of hsa-let-7b was associated with the tumor type
ABSTRACT
Background: Diagnosis of Non-small Cell Lung Cancer [NSCLC] at an early stage is a daunting challenge due to the deficiency of specific noninvasive markers. MicroRNAs [miRNAs] play important roles in the initiation and progression of NSCLC. Measuring miRNA expression levels could provide a potential approach for the diagnosis of NSCLC. Our goals were to examine miR-223, miR-212, miR-192, miR-3074, SNORD33 and SNORD37 expression levels in tissue and sputum of NSCLC patients and cancer free subjects for molecular diagnosis of NSCLC
Methods: Relative expressions of miR-223, miR-212, miR-192, miR-3074, SNORD33 and SNORD37 were examined with quantitative real-time RT-PCR assay in tissue and sputum obtained from 17 NSCLC patients and 17 controls
Results: miR-3074 was upregulated in tissue samples of NSCLC patients compared with control group. miR-223 was upregulated, miR-212 and SNORD37 were downergulated in sputum samples of patients compared with controls. miR-223 quantification produced 82% sensitivity and 95% specificity with areas under the ROC curve at 0.90 in detection of NSCLC
Conclusion: miR-223 clearly discriminated cancer patients from cancer-free subjects and our results suggest that miR-223 could be a diagnostic useful biomarker. The measurement of altered miRNA expression in sputum samples manifested the potential noninvasive approach for detection of lung cancer
ABSTRACT
Background: The human OCT4 gene, responsible for pluripotency and self-renewal of Embryonic Stem [ES] and Embryonic Carcinoma [EC] cells, can generate several transcripts [OCT4A, OCT4B-variant 2, OCT4B-variant 3, OCT4B-variant 5, OCT4B1, OCT4 B2 and OCT4B3] by alternative splicing and alternative promoters. OCT4A that is responsible for ES and EC cell stemness properties is transcribed from a promoter upstream of Exon1a in those cells. The OCT4B group variants [OCT4B-varianT[2], OCT4Bvariant3, OCT4B-variant5, OCT4B1, OCT4B2 and OCT4B3] are transcribed from a different promoter located in intron 1 and some of them respond to the cell stresses, but cannot sustain the ES/EC cell self-renewal. However, the exact function of OCT4B group variants is still unclear
Methods: In the present study, we employed RT-PCR and sequencing approaches to explore different forms of OCT4 transcripts
Results: Our data revealed that the OCT4B group variants [OCT4B-varianT[2], OCT4 B-variant3, OCT4B1, OCT4B2 and OCT4B3] have longer 5' UTR in the human bladder carcinoma cell line of 5637
Conclusion: These OCT4 variants undergo alternative splicing in their 5' UTR which might exert regulatory roles in transcription and translation mechanisms
ABSTRACT
Background: The objective of this study was determination of the changes in the reactive oxygen species [ROS] level, mitochondrial DNA [mtDNA] copy number and enzyme activity and transcription factor A [TFAM] gene expression in oocytes after vitrification
Methods: The oocytes at metaphase II [MII] stage [n=320] were collected from superovulated adult female mice [n=40]. These oocytes were divided into vitrified and non-vitrified groups [n=160 in each group]. After vitrification of oocytes, ROS level, mtDNA copy number; TFAM gene expression and mitochondrial enzymes activity [cytochrome C oxidase and succinate dehydrogenase] were assessed and compared with non-vitrified group. Visualization of the mitochondria was done using Mitotracker green staining under confocal microscope. Data were compared by independent T-test. Values of p<0.05 were considered as statistically significant
Results: The survival rate of oocytes after vitrification and warming was 96.05%. The intensity of cytochrome C oxidase activity, mtDNA copy number and TFAM gene expression in non-vitrified oocytes were significantly lower and the level of ROS was higher in vitrified oocytes in comparison with non-vitrified group [p<0.05]. But the intensity of succinate dehydrogenase activity was not significantly different between the two groups. The pattern of mitochondrial distribution in two groups of study was similar but the intensity of Mitotracker green in non-vitrified oocytes was significantly higher than vitrified oocytes [p<0.05]
Conclusion: This study showed that vitrification of mouse MII oocytes reduced the mtDNA copy number and mitochondrial cytochrome C oxidase activity by increasing ROS level, thus the subsequent embryo development may be affected
ABSTRACT
Background: Alternative splicing is an important mechanism that regulates gene expression and function in human cells. OCT4, a crucial pluripotency marker in embryonic stem/carcinoma cells generates several spliced variants in different cell types and cancers. The expression of OCT4 in cancers has been challenged in many studies. The existence of several OCT4 spliced variants and absence of specific discriminating primers is the main reason of this controversy. Therefore, using specific primers and discriminating OCT4 variants from each other might help to reduce these discrepancies in carcinogenesis and stem cell researches
Methods: 17 various human cancer, pluripotent and normal cells were cultured and their RNAs were extracted. Related cDNAs were synthesized and the expression pattern of OCT4variants was investigated by RT-PCR assay. PCR products were cloned into pTZ57R/T vector and their authenticity was confirmed by DNA sequencing
Results: Expression pattern of OCT4 variants [OCT4A, OCT4B and OCT4B1] was analyzed by RT-PCR assay and the authenticity of PCR products was confirmed by DNA sequencing. A novel spliced variant of OCT4 was discovered and named as OCT4B3. This variant was very similar to OCT4B2 transcript except that 207-nt of exon 1b is lost. Moreover, the expression pattern of OCT4B3 variant was investigated in 17 human cell types, where its expression was only found in astrocytoma and bladder cancer cell types 1321N1 and 5637, respectively
Conclusion: OCT4 variants are differentially expressed in various human cancer cell lines. Moreover, a novel variant of OCT4, OCT4B3, was detected in two human cancer cell lines of bladder carcinoma [5637] and brain astrocytoma [1321N1] for the first time
ABSTRACT
Objective: This study aimed to evaluate the expression of the genes related to folliculo-genesis after vitrification of mouse ovarian tissues using a two-step in vitro culture
Materials and Methods: In this experimental study, vitrified and non-vitrified ovaries from 7- day old [neonate] female mice were cultured using alpha-Minimum Essential Medium [alpha-MEM] supplemented with 5% fetal bovine serum [FBS] for 7 days. Morphology, surface area of ovaries and percentage of normal follicles were evaluated and compared in both groups. After one-week culture, in non-vitrified group, preantral follicles of cultured ovaries were isolated and cultured in a three-dimensional alginate culture system for 12 days. Then, the collected metaphase [M] II oocytes were inseminated with capacitated spermatozoa derived from 7-8-week old [adult] male NMRI mice. Follicular diameter, oocyte maturation, fertilization, embryo development and the expression of genes related to follicular development [Pcna, Fshr and Cyp17a1,] using real time reverse transcription-polymerase chain reaction [RT-PCR] were assessed at the end of last culture period in both groups
Results: The ovarian area in vitrified group [162468.20 703.78] was less than non-vitrified group [297211.40 6671.71], while the percentage of preantral follicles in vitrified group [18.40%] was significantly lower than those of non-vitrified group [24.50%] on day 7 of culture [P<0.05]. There were no significant differences between the two groups in terms of follicular diameter, expression of genes related to development of follicles, oocyte maturation, fertilization, as well as embryo development [P>0.05]
Conclusion: The results of this study showed that vitrification of ovarian tissue following in vitro culture had negative impact on the survival and development of follicles within the tissue. However, no significant alterations were observed in development, gene expression and hormonal production of in vitro culture of isolated follicles derived from vitrified ovarian tissues as compared to the non-vitrified samples
ABSTRACT
Objective: OCT4B1, a novel variant of OCT4, is expressed in cancer cell lines and tis-sues. Based on our previous reports, OCT4B1 appears to have a crucial role in regulating apoptosis as well as stress response [heat shock proteins [HSPs]] pathways. The aim of the present study was to determine the effects of OCT4B1 silencing on the expression of high molecular weight HSPs in three different human tumor cell lines
Materials and Methods: In this experimental study, OCT4B1 expression was suppressed in AGS [gastric adenocarcinoma], 5637 [bladder tumor] and U-87MG [brain tumor] cell lines using RNAi strategy. Real-time polymerase chain reaction [PCR] array was employed for expression level analysis and the fold changes were calculated using RT2 Profiler PCR array data analysis software version 3.5
Results: Our data revealed up-regulation of HSPD1 [from HSP60 family] as well as HSPA14, HSPA1L, HSPA4, HSPA5 and HSPA8 [from HSP70 family] following OCT4B1 knock-down in all three cell lines. In contrast, the expression of HSP90AA1 and HSP90AB1 [from HSP90 family] as well as HSPA1B and HSPA6 [from HSP70 family] was down-regulated under similar conditions. Other stress-related genes showed varying expression pattern in the examined tumor cell lines
Conclusion: Our data suggest a direct or indirect correlation between the expression of OCT4B1 and HSP90 gene family. However, OCT4B1 expression was not strongly correlated with the expression of HSP70 and HSP60 gene families