Effects of Long Noncoding RNA TUG1 (Taurine Up-Regulated Gene 1) on Growth and Metastasis of the Non-Small Cell Lung Cancer and Its Mechanism

Objective: Our research was to discuss effects and mechanism of lncRNA TUG1 in NSCLC by vitro study. Methods: A549 and H1299 cells were divided into NC, pcDNA 3.1 and lncRNA TUG1 groups. Measuring cell proliferation using CCK-8 assay, cell apoptosis by flow cytometry, invasion cell number by transwell and wound healing rate by wound healing assay. Relative gene and protein expressions by RT-qPCR and WB assay. Results: Compared with NC group, the cell proliferation rate, invasion cell number and wound healing rate were significantly depressed in A549 and H1299 cell lines (P < 0.001, respectively). By RT-qPCR and WB assay, lncRNA TUG1 gene expression were significantly increased (P < 0.001, respectively); E-cadherin gene and protein expression were significantly up-regulation, and N-cadherin and Vimentin gene and protein expressions were significantly depressed compared with those of NC group in A549 and H1299 cell lines (P < 0.001, respectively). Conclusion: lncRNA TUG1 had effects to suppress NSCLC cell biological activities by regulation EMT relative gene and proteins expression in vivo study.

TiO2 nanoparticles and salinity stress in relation to artemisinin production and ADS and DBR2 expression in Artemisia absinthium L.

Artemisia absinthium L. is an important herb that is widely cultivated in different parts of the world for its medicinal properties. The present study evaluated the effects of four concentrations of nanoparticles treatment (0, 10, 20 and 30 mg L-1) and NaCl salinity stress (0, 50, 100 and 150 mM NaCl) and their interactions with respect to the expression of two key genes, i.e. DBR2 and ADS, in the biosynthesis pathway of artemisinin in A. absinthium. Total RNA was extracted and a relative gene expression analysis was carried out using Real-Time PCR. The amount of artemisinin was also determined by HPLC. All the experiments were performed as factorial in a completely randomized design in three replications. The results revealed that salinity stress and nanoparticles treatment and their interaction affected the expressions of these genes significantly. The highest levels of ADS gene expression were observed in the 30 mg L-1 nanoparticles–treated plants in the presence of 150 mM salinity stress and the lowest levels in the 10 mg L-1 nanoparticles–treated plants under 50 mM salinity stress. The maximum DBR2 gene expression was recorded in the 10 mg L-1 nanoparticles–treated plants in the absence of salinity stress and the minimum expression in the 100 mM salinity-stressed plants in the absence of nanoparticles treatment. Moreover, the smallest amounts of artemisinin were observed in the 150 mM salinity-stressed plants in the absence of nanoparticles and the highest amounts in the 30 mg L-1 nanoparticles–treated plants. The maximum amounts of artemisinin and ADS gene expression were reported from the plants in the same nanoparticles treatment and salinity stress conditions. In this regard, the amount of artemisinin was decreased by half in the plants containing the highest DBR2 gene expression. Meanwhile, no significant correlation was observed between these gene expressions and the artemisinin amount in the other nanoparticles–treated plants under different levels of salinity stress. The biosynthetic pathway of secondary metabolites appears to be very complex and dose not directly dependent on these gene expressions.

Cold shock treatment alleviates chilling injury in peach fruit by regulating antioxidant capacity and membrane lipid metabolism

Objectives The work intended to reveal the effect of cold shock (CS) treatment on chilling injury (CI), antioxidant capacity, and membrane fatty acid of peach fruit. Materials and methods Peaches were soaked in ice water (0 °C) for 10 min and stored at 5 °C for 28 days for determination, except CI, and then stored for 3 days at 20 °C, only CI was measured. The electrolyte leakage (EL) was measured by conductivity meter. The activities of antioxidant enzymes (superoxide dismutase, ascorbate peroxidase, catalase, and peroxidase) and key enzymes of membrane lipid metabolism (phospholipase D, lipase, and lipoxygenase) as well as reactive oxygen species (ROS; O2·– and H2O2) were measured with a spectrophotometer. An ELISA kit and gas chromatography were used to determine membrane lipids and membrane fatty acids. The relative gene expression was measured by real-time polymerase chain reaction analysis. Results The results showed that CS treatment effectively delayed CI, suppressed the increase of EL and malondialdehyde content. Meanwhile, CS-treated fruit exhibited lower level of ROS and higher activities of antioxidant enzymes. Furthermore, CS treatment inhibited the activities as well as the relative gene expression of key enzymes in membrane lipid metabolism. CS-treated fruits maintained higher membrane fatty acid unsaturation and lower phosphatidic acid content. Conclusions These results indicated that CS treatment effectively alleviated CI and maintained the integrity of cell membranes by inducing antioxidant-related enzyme activity and maintaining a higher ratio of unsaturated fatty acids to saturated fatty acids.

Use of melatonin as an inhibitor of apoptotic process for cryopreservation of zebrafish (Danio rerio) embryos

This study investigated the use of melatonin to arrest the effects of apoptosis in vitrified zebrafish (D. rerio) embryos. Dechorionated embryos at 22-24 somite-stage were divided (n = 60/treatment) into a non-vitrified (Control Group, 0 M melatonin) and vitrified treatments with 0 M (T1), 1 µM (T2) and 1 mM of melatonin (T3). For vitrified treatments, a solution methanol/propylene glycol based was used and the embryos stored in -196 °C for a week. After thaw, survival rate, scanning electron microscopy, expression of anti (bcl-2) and pro-apoptotic (bax/caspase-3) genes, reactive oxygen species (ROS) formation and DNA fragmentation analyses were performed. No live embryos were obtained from vitrified treatments, observing a rapid degeneration immediately after thawing, with the vitelline layer rupture and leakage of its content, followed by breakdown of epithelial cells and melanisation of the tissue. Regarding the apoptotic process, T3 had the highest relative gene expression, for the three genes (P < 0.05) furthermore, T2 had similar expression of pro-apoptotic genes to CG (P < 0.05). ROS formation revealed that CG presented lower percentage of embryo surface area affected (3.80 ± 0.40%) (P < 0.05), in contrast, no differences were found among the other groups. T1 was most significantly (P < 0.05) damaged by DNA fragmentation. The vitrified groups with melatonin had similar damage levels of CG (P > 0.05). The inclusion of 1 µM of melatonin in the vitrifying solution, countered the effects of apoptotic process in post-thaw embryos, suggesting its utility in cryopreserving fish embryos.

Quinoa (Chenopodium quinoa Willd.): Genetic Diversity According to ISSR and SCoT Markers, Relative Gene Expression, and Morpho-Physiological Variation under Salinity Stress

Quinoa (Chenopodium quinoa Willd.) is a halophytic crop that can withstand a variety of abiotic stresses, including salt. The present research examined the mechanisms of salt tolerance in five different quinoa genotypes at four different salinity levels (control (60), 80, 120, and 160 mM NaCl). ISSR and SCoT analysis revealed high polymorphism percentages of 90.91% and 85.26%, respectively. Furthermore, ISSR 1 and SCoT 7 attained the greatest number of polymorphic amplicons (27 and 26), respectively. Notably, LINE-6 and M-28 genotypes demonstrated the greatest number of unique positive and negative amplicons (50 and 42) generated from ISSR and SCoT, respectively. Protein pattern analysis detected 11 bands with a polymorphism percentage 27.27% among the quinoa genotypes, with three unique bands distinguishable for the M-28 genotype. Similarity correlation indicated that the highest similarity was between S-10 and Regeolone-3 (0.657), while the lowest similarity was between M-28 and LINE-6 (0.44). Significant variations existed among the studied salinity treatments, genotypes, and the interactions between them. The highest and lowest values for all the studied morpho-physiological and biochemical traits were recorded at 60 and 160 mM NaCl concentrations, respectively, except for the Na and proline contents, which exhibited the opposite relationship. The M-28 genotype demonstrated the highest values for all studied characteristics, while the LINE-6 genotype represented the lowest in both seasons. On the other hand, mRNA transcript levels for CqSOS1 did not exhibit differential expression in roots and leaf tissues, while the expression of CqNHX1 was upregulated more in both tissues for the M-28 genotype than for the LINE-6 genotype, and its maximum induction was seen in the leaves. Overall, the genotypes M-28 and LINE-6 were identified as the most and least salinity-tolerant, respectively.

Determination of Genetic Diversity Based on RAPD molecular Marker and ParARF3 Gene Expressions in some Apricot Genotypes in Iraq

Employing DNA markers allowed determining genetic diversity and relationships amongst five apricot genotypes. In this study, two relative gene expressions pertaining to ParARF3 gene, which could be distinguished from the genotypes that were exposed to various concentrations of marine alga treatments. As per our results, screening of seven primers with the DNA of 5 apricot genotypes was done, and six primers were generated while the primer OPN–16 gave negative results. The total quantity of fragments generated by 6 primers was 80 at an average of 13.33 fragments ̸primer. The highest unique percentage band depicted in U-17 touched 23%, and the total number of polymorphic bands touched 17 fragments with the average reaching 2.83 fragments ̸primer. The number of monomorphic lied in the range of 5 to 10, with a total of 47 monomorphic. Primer M 32 yielded the highest number of monomorphic bands reaching 10. Between Zaghenia and Zinni, a maximum genetic distance value of 0.8 was reached with less similarity value of 20%. A minimum genetic distance value of 0.44721 was noted between Kaisy and Baia while the high similarity value touched 55.3%. According to the cluster tree analysis, the genotypes were generally split into two key groups: A and B. The Zinni group, which included one apricot genotype, showed genetic similarity of 20% with the other genotypes present in B group. The B group was split into two sub-clusters B1 and B2 and the genetic similarity reached 44%. The ParARF3 relative gene expression pertaining to Zinni genotypes was second as well as convergent with that of gene expression for Zaghenia genotype results. Baia and Kaisy genotypes lied in between the lowest and highest ParARF3 value gene expression exposed to Marine Alga. These outcomes showed that RAPD markers offer an effectual alternative for the plant species genetic characterisation.

A 4-Gene Signature of CDKN1, FDXR, SESN1 and PCNA Radiation Biomarkers for Prediction of Patient Radiosensitivity

The quest for the discovery and validation of radiosensitivity biomarkers is ongoing and while conventional bioassays are well established as biomarkers, molecular advances have unveiled new emerging biomarkers. Herein, we present the validation of a new 4-gene signature panel of CDKN1, FDXR, SESN1 and PCNA previously reported to be radiation-responsive genes, using the conventional G2 chromosomal radiosensitivity assay. Radiation-induced G2 chromosomal radiosensitivity at 0.05 Gy and 0.5 Gy IR is presented for a healthy control (n = 45) and a prostate cancer (n = 14) donor cohort. For the prostate cancer cohort, data from two sampling time points (baseline and Androgen Deprivation Therapy (ADT)) is provided, and a significant difference (p > 0.001) between 0.05 Gy and 0.5 Gy was evident for all donor cohorts. Selected donor samples from each cohort also exposed to 0.05 Gy and 0.5 Gy IR were analysed for relative gene expression of the 4-gene signature. In the healthy donor cohort, there was a significant difference in gene expression between IR dose for CDKN1, FXDR and SESN1 but not PCNA and no significant difference found between all prostate cancer donors, unless they were classified as radiation-induced G2 chromosomal radiosensitive. Interestingly, ADT had an effect on radiation response for some donors highlighting intra-individual heterogeneity of prostate cancer donors.

Oocyte Morphometric Assessment and Gene Expression Profiling of Oocytes and Cumulus Cells as Biomarkers of Oocyte Competence in Sheep

Oocyte quality is crucial for subsequent embryo development and so it is a major challenge in assisted reproductive technologies. The aim of the present work was to evaluate the morphometric parameters of oocytes (experiment 1) and the relative gene expression of oocytes and cumulus cells (CCs) (experiment 2) as biomarkers of oocyte quality after individually culturing them (one oocyte or embryo/drop). In experiment 1, individually matured oocytes were measured and classified into small, intermediate, and large oocytes after a cluster analysis, based on total diameter (with zona pellucida, ZP), oocyte diameter (without ZP), and ZP thickness. These oocytes were individually fertilized in vitro and cultured. The embryo development was evaluated up to the blastocyst stage. According to the total diameter, oocyte diameter, and ZP thickness, the blastocyst rate decreased in the small oocytes group (3.1 ± 3.1, 14.1 ± 9.4, and 26.7 ± 3.9, respectively) compared to the intermediate (29.4 ± 5.2, 30.5 ± 10.1, and 28.6 ± 9.6, respectively) and large oocytes groups (54.2 ± 13.5, 44.4 ± 3.9, and 67.6 ± 12.4, respectively). In addition, the probability of reaching the blastocyst stage was positively related to the total diameter (p < 0.001), oocyte diameter (p < 0.05), and ZP thickness (p < 0.001). Furthermore, the relative gene expression of BAX, BCL2, GDF9, and GJA1 was lower in oocytes classified as large. In experiment 2, the mRNA transcript relative abundance pattern of genes in CCs was evaluated according to oocyte total diameter and developmental stage reached. CCs from oocytes classified as large and oocytes capable of developing to the blastocyst stage had a lower relative expression of BAX, STAR, and PTGS2, while a higher expression of HAS2 and SDC2 transcript was observed for those oocytes. In conclusion, oocyte morphometric parameters and gene expression analysis in oocytes and CCs provide methods for the identification of the most competent oocytes for assisted reproductive technologies in sheep.