Mina Lashkarboloki; Amin Jahanbakhshi; Seyed Javad Mowla; Bahram Mohammad Soltani
Abstract
Cancer is one of the most challenging diseases in the world. It is widely accepted that knowing the molecular aspects of diseases, including cancers, helps to develop methods for their therapy and diagnosis. Long non-coding RNAs (lncRANs) are a novel category of regulatory genes known to be involved ...
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Cancer is one of the most challenging diseases in the world. It is widely accepted that knowing the molecular aspects of diseases, including cancers, helps to develop methods for their therapy and diagnosis. Long non-coding RNAs (lncRANs) are a novel category of regulatory genes known to be involved in cancer incidence. The expression of these genes is said to be suitable of using in prognosis, diagnosis, targeted therapy, etc. The RT-qPCR method that is widely used for analyzing the gene expression requires the application of appropriate reference genes as the internal control. The expression status of a proper housekeeping reference gene is not supposed to change under experimental circumstances. This study aimed to find a suitable reference gene in the U87 cells after overexpression of a gene of interest. To this aim, the expression status of four common reference genes (ACTB, β2M, GAPDH, and HPRT1) was examined in the transfected U87 cells. The U87 cells were transfected with a vector overexpressing YWHAE-lncRNA and an empty vector (mock). After total RNA extraction and cDNA synthesis, RT-qPCR was applied using the aforementioned internal control genes. Data were analyzed, and their graphs were plotted in GraphPad Prism 8.2 software. Β2M showed the most change; accordingly, GAPDH and HPRT1 expression levels were changed about 5 and 4 times, respectively. Of the candidate genes, only the ACTB gene had a consistent expression level in two different modes of transfection, and therefore, it is suggested as an appropriate reference gene for the study of gene expression in the transfected U87 cell line. It is remained to be tested if β2M, GAPDH, and HPRT1 common internal controls are specifically affected by YWHAE-lncRNA overexpression or other lncRNAs may affect their expression as well.
Mohammadreza Nassiri; Azadeh Safarchi; Masoume Vakili-Azghandi; Vinod Gopalan; Mohammad Doosti; Shahrokh Ghovvati; Ahmad Reza Movassaghi
Abstract
p53 is a tumor suppressor protein that plays an essential role in controlling the cell and vascular endothelial growth factor (VEGF) is one of the most strong and specific angiogenic factors. The main objective of this study was to evaluate the impact of p53 and VEGF-C gene expression in the neoplastic ...
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p53 is a tumor suppressor protein that plays an essential role in controlling the cell and vascular endothelial growth factor (VEGF) is one of the most strong and specific angiogenic factors. The main objective of this study was to evaluate the impact of p53 and VEGF-C gene expression in the neoplastic and normal mammary gland of canine as an animal model. Elleven benign and malignant specimens and 5 normal specimens were collected. After RNA extraction and cDNA synthesis, relative quantification of p53 and VEGF-C genes were accomplished by Real-time quantitative PCR (RT-qPCR) based on use of β-actin as a reference gene. The relative mRNA expression of the p53 and VEGF-C genes were analyzed by GLM procedure of SAS software v9.2. The results indicated that the VEGF-C and p53 mRNA expression in neoplastic specimens was over-and down-expressed respectively as compared with normal specimens and p53 mRNA expression was significantly negatively associated with VEGF-C (~4 fold) in neoplastic specimens (P <0.01). The findings emphasized that simultaneous evaluation of p53 and VEGF-C expression can be used as tumor biomarker for early diagnosis of malignancy in canine. Furthermore, RT-qPCR is a rapid and sensitive method to for monitoring and investigating of suspicious canine at the beginning stage of malignancy and may provide an alternative explanation for deregulated p53 signalling in breast cancer.
Jafar Vatandoost; Shadi Tirdad
Abstract
In the production of recombinant proteins, the selection of an expression system is very important. Although CHO cells are used as mammalian expression system, the ability of insect Schneider line 2 (S2) cells as a new expression system for the high production of many human proteins were confirmed. It ...
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In the production of recombinant proteins, the selection of an expression system is very important. Although CHO cells are used as mammalian expression system, the ability of insect Schneider line 2 (S2) cells as a new expression system for the high production of many human proteins were confirmed. It is suggested that high copy number of an introduced gene and so high transcription in these cells can be regarded as one of the reasons for high expression of recombinant proteins. Therefore, the present study aimed to evaluate the correlation of recombinant human coagulation factor IX (hFIX) abundance and mRNA expression. The amount of hFIX mRNA by quantitative real-time PCR as well as hFIX protein in cell lysate and cultured media by Elisa was analyzed. The results of data analysis indicated 6 fold increases in mRNA level in S2 cells in comparison to CHO cells. Furthermore, S2 cell line indicated 5.5 and 7-fold increase in total and secreted protein level, respectively, compared to CHO cell line. The data demonstrated the correlation of mRNA and protein abundance and indicate that S2 cell lines are superior in producing the recombinant proteins.
fatemeh keykha; Abdol Reza Bagheri; Nasrin Moshtaghi
Abstract
Variegation in flower color is commonly observed in many plant species and also occurs on petunia (Petunia hybrida)
as an ornamental plant. Variegated plants are highly valuable in the floricultural market. To gain a global perspective
on genes differentially expressed in variegated petunia flowers, ...
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Variegation in flower color is commonly observed in many plant species and also occurs on petunia (Petunia hybrida)
as an ornamental plant. Variegated plants are highly valuable in the floricultural market. To gain a global perspective
on genes differentially expressed in variegated petunia flowers, we investigated the expression of chalcone synthase
(chs) and chalcone isomerase (chi) as two essential genes in biosynthesis pathway of pigment production. Also, we
measured the concentration level of total flavonoids, naringenin chalcone and naringenin to evaluate the probably
relationship between the expression profile of chs and chi genes and the concentration of mentioned pigments. The
results indicated that chalcone synthase and chalcone isomerase expression had different profile in different petal
color of Petunia hybrida. Because red flower color in petunia is related to the synthesis of pelargonidin-based (orange
to red) pigments, our results suggest that the low chalcone synthase and chalcone isomerase expression levels in
white petals reduce dihydrokaempferol formation, thereby inhibiting pelargonidin production. In contrast, the high
expression levels of these genes observed in red petals ensure sufficient anthocyanin yields to make flowers red.
Maria Beihaghi; Ahmad Reza Bahrami; Abdolreza Bagheri; Mohammad Zare Mehrjerdi
Abstract
Studies on the genes contributing to the seed filling in chickpea and its protein content might be valuable in engineering plants with seeds of a higher nutritional value. A gene of interest is phosphoenolpyruvate carboxykinase (pepck), encoding a protein with a substantial role in the gluconeogenesis ...
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Studies on the genes contributing to the seed filling in chickpea and its protein content might be valuable in engineering plants with seeds of a higher nutritional value. A gene of interest is phosphoenolpyruvate carboxykinase (pepck), encoding a protein with a substantial role in the gluconeogenesis pathway. In the present study, the protein content (percentage) was measured in a number of cultivated chickpea genotypes, followed by comparison of the expression levels of pepck gene at different stages of seed filling in some of the genotypes. This study aims at revealing the relation between pepck transcript level with protein content of chickpea seeds which might, in the longer term, end at protein quality improvement of the crop through the gene manipulation procedures. The results which were verified by Real Time-PCR and Western blot techniques in four genotypes of plant, showed that, the amount of pepck expression was significantly higher at the stage of seed fillingthan at other stages in all of genotypes and the lowest levels of expression belonged to flowering and seed formation and the PEPCK protein level was higher in the high protein genotypes compared to the low protein genotypes.
Nasrin moshtaghi; Robab Ghahremanzadeh; Seyyed Hasan Marashi
Abstract
Saffron (Crocus sativus L.) is an indigenous and the most valuable and marginal plant in Iran. However,
limited knowledge exists on its molecular biology. The importance of this plant is due to the color, flavor and
medicinal properties of its red stigmas. Saffron stigmas contain a high amount of carotenoids ...
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Saffron (Crocus sativus L.) is an indigenous and the most valuable and marginal plant in Iran. However,
limited knowledge exists on its molecular biology. The importance of this plant is due to the color, flavor and
medicinal properties of its red stigmas. Saffron stigmas contain a high amount of carotenoids such as crocetin
and crocin. Two genes, bch and pds, have essential role in carotenoid production. In this study, the effect of
four different irrigation regimes was evaluated on the expression of bch and pds genes. Semi-quantitative RTPCR
showed no significant difference in the expression levels of genes of interest related to the internal
standard (18S rRNA). Results of Real-Time PCR assays showed that the expression of bch and pds genes were
affected by irrigation treatments as their expression decreased in irrigated plants in comparison to non irrigated
ones, exept for one irrigation treatment (one irrigation in September) where the pds gene expression showed
higher level. However, the expression profile of the genes was almost the same in all treatments. The
comparison between results of two techniques indicated that the Real-Time PCR is more accurate for
determination of the level of transcript in the Iranian saffron. It was interesting that by decreasing of irrigation,
the expression level of these two genes increased indicating that abiotic stress and drought can affect on the
gene expression relating to the saffron color.
