Journal of Biosciences

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PEGylated Lipova E120 liposomes loaded with celecoxib: in-vitro characterization and enhanced in-vivo anti-inflammatory effects in rat models
(Indian Academy of Sciences, 2019-09) Dave, Vivek; Gupta, Ashish; Singh, Priyanka; Tak, Kajal; Sharma, Swapnil
Brown adipose tissue (BAT) is responsible for adaptive thermogenesis. We previously showed that genetic deficiency ofreceptor for advanced glycation end products (RAGE) prevented the effects of high-fat diet (HFD). This study was tocompare BAT activity in RAGE knock out (Ager-/-, RKO) and wild-type (WT) mice after treated with HFD or LFD.[18F]FDG PET-CT imaging under identical cold-stimulated conditions and mean standard uptake values (SUVmean), ratio ofSUViBAT/SUVmuscle (SUVR, muscle as the reference region) and %ID/g were used for BAT quantification. The resultsshowed that [18F]FDG uptake (e.g., SUVR) in WT-HFD mice was significantly reduced (three-fold) as compared to that inWT-LFD (1.40 ± 0.07 and 4.03 ± 0.38; P = 0.004). In contrast, BAT activity in RKO mice was not significantly affectedby HFD, with SUVRRKO-LFD: 2.14 ± 0.10 and SUVRRKO-LFD: 1.52 ± 0.13 (P = 0.3). The uptake in WT-LFD was almostdouble of that in RKO-LFD (P = 0.004); however, there was no significant difference between RKO-HFD and WT-HFDmice (P = 0.3). These results, corroborating our previous findings on the measurement of mRNA transcripts for UCP1 inthe BAT, suggest that RAGE may contribute to altered energy expenditure and provide a protective effect against HFD byAger deletion (Ager -/-).
Calycosin down-regulates c-Met to suppress development of glioblastomas
(Indian Academy of Sciences, 2019-09) Nie, Xiaohu; Zhou, Yue; Li, Xiaobing; Xu, Jie; Pan, Xuyan; Yin, Rui; Lu, Bin
The current case–control study sought the association of BDNF rs6265 and MC4R rs17782313 with metabolic syndrome(MetS), MetS components and other related metabolic parameters in a sample of Pakistani subjects. Fasting high-densitylipoprotein cholesterol (HDL-C) and homeostatic model assessment of insulin sensitivity showed a significantly lowermean whereas body mass index (BMI), waist circumference, systolic blood pressure (SBP), diastolic blood pressure (DBP),fasting blood glucose, insulin, total cholesterol (TC), low-density lipoprotein cholesterol, very-low-density lipoproteincholesterol, triglycerides (TG), cholesterol to HDL-C ratio, TG to HDL-C ratio, homeostatic model assessment of insulinresistance, visceral adiposity index, lipid accumulation product and the product of TG and glucose showed a significantlyhigher mean in the presence of MetS. Reduced HDL-C appeared as the most frequent and hypertriglyceridemia as the leastfrequent component of MetS whereas clustering of reduced HDL-C ? abdominal obesity (AO) ? hyperglycemia appearedas the most prevalent combination of MetS components. Moreover, BDNF rs6265 showed BMI and gender independentassociation with increased risk of MetS in Pakistani individuals whereas MC4R rs17782313 showed BMI and genderdependent association with increased risk of MetS in Pakistani females. In addition, BDNF rs6265 and MC4R rs17782313showed gender-dependent associations with decreased risk of having low HDL-C in males and increased risk of havingabdominal obesity in females, respectively. However, no association was observed for metabolic variables other thancomponents of MetS across genotypes of both BDNF rs6265 and MC4R rs17782313.
Association of BDNF rs6265 and MC4R rs17782313 with metabolic syndrome in Pakistanis
(Indian Academy of Sciences, 2019-09) Rana, Sobia; Sultana, Ayesha; Bhatti, Adil Anwar
The goal of the current investigation was to prepare PEGylated Lipova E120 liposomes loaded with celecoxib for theeffective treatment of rheumatoid arthritis (RA). PEGylated liposomes were prepared and were characterized using techniques such as particle size distribution, polydispersity index (PDI), zeta potential, encapsulation efficiency and in-vitrorelease, in-vivo and stability studies. The morphological study was characterized by scanning electron microscopy andtransmission electron microscopy. To determine the interaction between drug and polymer Fourier transform infrared,Raman, thermogravimetric analysis and differential scanning calorimetry studies were performed. Results show that formulation F6 was optimized with a particle size of 92.12 ± 1.7 nm, a PDI of 0.278 ± 0.22, a zeta potential of- 40.8 ± 1.7 mV with a maximum encapsulation of 96.6 ± 0.05% of drug in the PEGylated liposomes. The optimizedformulation shows a maximum release of drug i.e. 94.45 ± 1.13% in 72 h. Tail immersion assay shows that the optimizedformulation F6 significantly increases the reaction time and carrageenan-induced assay shows that the optimized formulation inhibits the increase in paw edema thus providing a pain relief treatment in RA. These results suggest that thePEGylated liposomes provide a sustained release of celecoxib and helps in effective treatment of RA.
Analysis of genome-wide SNPs based on 2b-RAD sequencing of pooled samples reveals signature of selection in different populations of Haemonchus contortus
(Indian Academy of Sciences, 2019-09) Khan, Sawar; Zhao, Xiaochao; Hou, Yini; Yuan, Chunxiu; Li, Yumei; Luo, Xiaoping; Liu, Jianzhi; Feng, Xingang
The antitumor effect of calycosin has been widely studied, but the targets of calycosin against glioblastomas are stillunclear. In this study we focused on revealing c-Met as a potential target of calycosin suppressing glioblastomas. In thisstudy, suppressed-cell proliferation and cell invasion together with induced-cell apoptosis appeared in calycosin-treatedU251 and U87 cells. Under treatment of calycosin, the mRNA expression levels of Dtk, c-Met, Lyn and PYK2 wereobserved in U87 cells. Meanwhile a western blot assay showed that c-Met together with matrix metalloproteinases-9(MMP9) and phosphorylation of the serine/threonine kinase AKT (p-AKT) was significantly down-regulated by calycosin.Furthermore, overexpressed c-Met in U87 enhanced the expression level of MMP9 and p-AKT and also improved cellinvasion. Additionally, the expression levels of c-Met, MMP9 and p-AKT were inhibited by calycosin in c-Met overexpressed cells. However, an AKT inhibitor (LY294002) only effected on MMP9 and p-AKT, not on c-Met. These datacollectively indicated that calycosin possibility targeting on c-Met and exert an anti-tumor role via MMP9 and AKT.
Impact of Pseudomonas putida RRF3 on the root transcriptome of rice plants: Insights into defense response, secondary metabolism and root exudation
(Indian Academy of Sciences, 2019-09) Kandaswamy, Rekha; Ramasamy, Mohan Kumar; Palanivel, Rameshthangam; Balasundaram, Usha
The parasitic nematode Haemonchus contortus is one of the world’s most important parasites of small ruminants that causessignificant economic losses to the livestock sector. The population structure and selection in its various strains are poorlyunderstood. No study so far compared its different populations using genome-wide data. Here, we focused on differentgeographic populations of H. contours from China (Tibet, TB; Hubei, HB; Inner Mongolia, IM; Sichuan, SC), UK andAustralia (AS), using genome-wide population-genomic approaches, to explore genetic diversity, population structure andselection. We first performed next-generation high-throughput 2b RAD pool sequencing using Illumina technology, andidentified single-nucleotide polymorphisms (SNPs) in all the strains. We identified 75,187 SNPs for TB, 82,271 for HB,82,420 for IM, 79,803 for SC, 83,504 for AS and 78,747 for UK strain. The SNPs revealed low-nucleotide diversity (p =0.0092–0.0133) within each strain, and a significant differentiation level (average Fst = 0.34264) among them. Chinesepopulations TB and SC, along with the UK strain, were more divergent populations. Chinese populations IM and HBshowed affinities to the Australian strain. We then analysed signature of selection and detected 44 (UK) and 03 (AS) privateselective sweeps containing 49 and 05 genes, respectively. Finally, we performed the functional annotation of selectivesweeps and proposed biological significance to signature of selection. Our data suggest that 2b-RAD pool sequencing canbe used to assess the signature of selection in H. contortus.
miR-183-5p enhances the radioresistance of colorectal cancer by directly targeting ATG5
(Indian Academy of Sciences, 2019-09) Zheng, Sheng; Zhong, Yong-Fu; Tan, De-Ming; Xu, Yue; Chen, Huai-Xiang; Wang, Dan
Intracellular trafficking is a field that has been intensively studied for years and yet there remains much to be learned. Part ofthe reason that there is so much obscurity remaining in this field is due to all the pathways and the stages that define cellulartrafficking. One of the major steps in cellular trafficking is fusion. Fusion is defined as the terminal step that occurs when acargo-laden vesicle arrives at the proper destination. There are two types of fusion within a cell: homotypic and heterotypicfusion. Homotypic fusion occurs when the two membranes merging together are of the same type such as vacuole tovacuole fusion. Heterotypic fusion occurs when the two membranes at play are of different types such as when anendosomal membrane fuses with a Golgi membrane. In this review, we will focus on all the protein components – Rabs,Golgins, Multisubunit tethers, GTPases, protein phosphatases and SNAREs – that have been known to function in both ofthese types of fusion. We hope to develop a model of how all of these constituents function together to achieve membranefusion. Membrane fusion is a biological process absolutely necessary for proper intracellular trafficking. Due to the degreeof importance multiple proteins are required for it to be properly carried through. Whether we are talking about heterotypicor homotypic fusion, any defects in the fusion machinery can result in disease states such as Parkinson’s and Alzheimer’sdisease. Although much research has significantly expanded our knowledge of fusion, there is still much more to belearned.
Analysis of chickpea gene co-expression networks and pathways during heavy metal stress
(Indian Academy of Sciences, 2019-09) Yadav, Birendra Singh; Srivastava, Swati Singhsameer; Mani, Ashutosh
Pseudomonas putida is widely used as a biocontrol agent, however, mechanisms by which it initiates the plants’ defenseresponse remains obscure. To gain an insight into the molecular changes that occur in plants upon plant growth-promotingrhizobacteria colonization, root transcriptome analysis by using a microarray was performed in rice using P. putida RRF3 (arice rhizosphere isolate). Data analysis revealed a differential regulation of 61 transcripts (48 h post-treatment), of which,majority corresponded to defense response, cell wall modification and secondary metabolism. Seven genes encodingsalicylic acid (SA) responsive pathogenesis-related proteins were up-regulated significantly (fold change ranges from 1 to4), which suggests that RRF3 has a profound impact on a SA-mediated defense signaling mechanism in rice. Investigationsperformed at later stages of RRF3 colonization by real-time polymerase chain reaction and high-performance liquidchromatography (HPLC) analysis confirmed the above results, demonstrating RRF3 as a potent biocontrol agent. Further,the impact of RRF3 colonization on root exudation, in particular, exudation of SA was investigated by HPLC. However,analysis revealed RRF3 to have a negative impact on root exudation of SA. Overall, this study shows that P. putida RRF3immunizes the rice plants by re-organizing the root transcriptome to stimulate plant defense responses (‘priming’), andsimultaneously protects itself from the primed plants by altering the rhizosphere chemical constituents.
PET imaging study of brown adipose tissue (BAT) activity in mice devoid of receptor for advanced glycation end products (RAGE)
(Indian Academy of Sciences, 2019-09) Ding, Yu-Shin; Malik, Noeen; Mendoza, Sebastian; Tuchman, Daniel; Pozo, Carmen Hurtado Del; Diez, Raquel Lopez; Schmidt, Ann-Marie
Does cartilageER-alpha overexpression correlate with osteoarthritic chondrosenescence? Indications from Labisia pumila OA mitigation
(Indian Academy of Sciences, 2019-09) Madzuki, Iffah Nadhira; Lau, Seng Fong; Shalan, Nor Aijratul Asikin Mohamad; Ishak, Nur Iliyani Mohd; Mohamed, Suhaila
Crop productivity and yield are adversely affected by abiotic and biotic stresses. Therefore, finding out the genesresponsible for stress tolerance is a significant stride towards crop improvement. A gene co-expression network is apowerful tool to detect the most connected genes during heavy metal (HM) stress in plants. The most connected genes maybe responsible for HM tolerance by altering the different metabolic pathways during the biotic and abiotic stress. In thesame line we have performed the GSE86807 microarray analysis of chickpea during exposure to chromium, cadmium andarsenic and analyzed the data. Common differentially expressed genes (DEGs) during exposure to chromium, cadmium andarsenic were identified and a co-expression network study was carried out. Hub and bottleneck genes were explored on thebasis of degree and betweenness centrality, respectively. A gene set enrichment analysis study revealed that genes likehaloacid dehydrogenase, cinnamoyl CoA reductase, F-box protein, GDSL esterase lipase, cellulose synthase, b-glucosidase13 and isoflavone hydroxylase are significantly enriched and regulate the different pathways like riboflavin metabolism,phenyl propanoid biosynthesis, amino acid biosynthesis, isoflavonoid biosynthesis and indole alkaloid biosynthesis.
The inner workings of intracellular heterotypic and homotypic membrane fusion mechanisms
(Indian Academy of Sciences, 2019-09) Cruz, Mariel Delgado; Kim, Kyoungtae
The glycolytic enzyme enolase of Staphylococcus aureus is a highly conserved enzyme which binds to human plasminogenthereby aiding the infection process. The cloning, over expression and purification of S. aureus enolase as well as the effectof various metals upon the catalytic activity and structural stability of the enzyme have been reported. The recombinantenzyme (rSaeno) has been purified to homogeneity in abundant amounts (60 mg/L of culture) and the kinetic parameters(Km = 0.23 ± 0.013 9 10-3 M; Vmax = 90.98 ± 0.00052 U/mg) and the optimum pH were calculated. This communication further reports that increasing concentrations of Na? ions inhibit the enzyme while increasing concentrations of K?ions were stimulatory. In case of divalent cations, it was found that Mg2? stimulates the activity of rSaeno while the rest ofthe divalent cations (Zn2?, Mn2?, Fe2?, Cu2?, Ni2? and Ca2?) lead to a dose-dependent loss in the activity with a total lossof activity in the presence of Hg2? and Cr2?. The circular dichroism data indicate that other than Hg2?, Ni2? and to acertain extent Cu2?, none of the other ions destabilized rSaeno. The inhibitory roles of fluorides, as well as neurotoxiccompounds upon the catalytic activity of rSaeno, have also been studied. Conformational changes in rSaeno (induced byions) were studied using partial trypsin digestion.
Comprehensive transcriptomics and proteomics analyses of rice stripe virus-resistant transgenic rice
(Indian Academy of Sciences, 2019-09) Xu, Yang; Bi, Lvjie; Yu, Zipeng; Lin, Chao; Gan, Liming; Zhu, Lifei; Li, Haiyang; Song, Yunzhi; Zhu, Changxiang
Stable transgenic rice line (named KRSV-1) with strong resistance against rice stripe virus was generated using the genesequence of disease-specific protein by RNA interference. Comprehensive safety assessment of transgenic plants has turnedinto a significant field of genetic modification food safety. In this study, a safety assessment of KRSV-1 was carried out in astepwise approach. The molecular analysis exhibited that KRSV-1 harbored one copy number of transgene, which wasintegrated into the intergenic non-coding region of chromosome 2 associated with inter-chromosomal translocations of 1.6-kb segments of chromosome 8. Then, transcriptomics and proteomics analyses were carried out to detect the unintendedeffects as a result of the integration of the transgene. Although 650 dramatically differentially expressed genes (DDEGs)and 357 differentially expressed proteins were detected between KRSV-1 and wild-type (WT) by transcriptomics andproteomics analyses, no harmful members in the form of toxic proteins and allergens were observed. Encouragingly, thenutritional compositions of seeds from KRSV-1 were comparable with WT seeds. The results of this entire study ofmolecular analysis, transcriptome and proteome profile of KRSV-1 revealed that no detrimental changes in the form of toxicproteins and allergens were detected in the transgenic rice line due to the integration of the transgene.
Effect of ions and inhibitors on the catalytic activity and structural stability of S. aureus enolase
(Indian Academy of Sciences, 2019-09) Hemmadi, Vijay; Das, Avijit; Chouhansumit, Om Prakash; Biswas, Biswasmalabika
Constantly rising energy demands, finite fossil fuel reserves and deteriorating environmental conditions have invokedworldwide interest to explore the sustainable sources of renewable biofuels. Locally adapted photosynthetic oleaginousmicroalgae with rapid growth on variable temperatures could be an ideal way for bioremediating the wastewater (WW) whileproducing the feedstock for biodiesel. To test this notion, an unknown strain was isolated from a sewage fed lake (Neela-Hauz).It was discerned as Chlorella sorokiniana-I using the 16S rDNA and 18S rDNA barcodes. The culture conditions such as pH,illumination, different temperature ranges and growth medium were cohesively optimized prior to the assessment of C.sorokiniana-I’s efficacy to remediate the WW and biodiesel production. The strain has thrived well up to 40C when continuously grown for 15 days. The highest lipid accumulation and biomass productivity were recorded in 100% WW. Fatty acidmethyl ester (FAME) content was observed to be more than twice in WW (47%), compared to control synthetic media, TAP(20%) and BG11 (10%), which indicate the importance of this new isolate for producing economically viable biodiesel.Moreover, it is highly efficient in removing the total nitrogen (77%), total phosphorous (81%), iron (67%) and calcium (42%)from the WW. The quality of WW was considerably improved by reducing the overall chemical oxygen demand (48%),biological oxygen demand (47%) and alkalinity (15%). Thus, C. sorokiniana-I could be an ideal alga for the tropical countriesin the remediation of WW while producing feedstock for biodiesel in a cost-effective manner.
Adiponectin inhibits vascular smooth muscle cell calcification induced by beta-glycerophosphate through JAK2/STAT3 signaling pathway
(Indian Academy of Sciences, 2019-09) Lu, Yan; Ma, Yichao; Wang, Ruihua; Sun, Jing; Guo, Beibei; Wei, Ruipeng; Jia, Yongping
Radioresistance is a material obstacle for effective treatment of colorectal cancer (CRC). Thus, the discovery of a novelbiomarker for determining the CRC radiosensitivity is necessary. Recent studies have confirmed that miR-183-3p regulatescell phenotypes and tumor growth in various cancers. However, the role and mechanism of this micro-ribonucleic acid inCRC radiosensitivity remains unclear. Here, the abundances of miR-183-5p and ATG5 mRNA were detected by a real-timequantitative reverse transcription polymerase chain reaction. Kaplan–Meier survival analysis was carried out to explore thecorrelation between miR-183-5p and patient prognosis. Cell viability was evaluated by the MTT assay. Survival fractionanalysis through colony formation was performed to assess the cell radiation response. Bioinformatic, luciferase andwestern blot assays were employed to verify the targeted interaction between miR-183-5p and ATG5. The results showedthat an elevated abundance of miR-183-5p and a reduced ATG5 level in CRC were associated with the poor prognosis. Theknockdown of miR-183-5p enhanced the sensitivity of CRC cells to radiation, inflected by the decreased cell viability andsurvival fraction. Mechanically, ATG5 was targeted by miR-183-5p. The addition of ATG5 conferred the radiosensitivity ofthe CRC cells, which was revered by miR-183-5p restoration. Furthermore, miR-183-5p knockdown hindered the tumorgrowth by repressing ATG5 in vivo after radiation treatment. In summary, the output data indicated that miR-183-5pheightened the radiation response of the CRC cells by targeting ATG5, promising a novel therapeutic target for CRCpatients with radioresistance.
Stability analysis of a prey refuge predator–prey model with Allee effects
(Indian Academy of Sciences, 2019-09) Ufuktepe, Unal; Akman, Burcin Kulahciogluolcay
We constructed a discrete-time predator–prey model by adding prey refuge and Allee effects (predator saturation on preyand mate limitation on predator) to an earlier prey–predator model and examined its dynamics. We show the existence ofpositive fixed points and study the stability properties. The numerical simulations and bifurcation diagrams verify theimpact of refuge and the Allee mechanism on the system.
Real-time monitoring of translocation of selected type-III effectors from Xanthomonas oryzae pv. oryzae into rice cells
(Indian Academy of Sciences, 2019-09) Huijie, Bian; Wang, Liyuan Zhanglei Chenwenzhan; Ji, Hongtao; Dong, Hansong
Type-III (T3) effectors PthXo1 and AvrXa10 of Xanthomonas oryzae pv. oryzae are translocated into rice cells to inducevirulence and avirulence on susceptible- and resistant-rice varieties Nipponbare and IRBB10, respectively. The translocation needs the bacterial T3 translocator Hpa1 and rice Oryza sativa plasma membrane protein OsPIP1;3. Here, weemployed the b-lactamase (BlaM) reporter system to observe PthXo1 and AvrXa10 translocation. The system wasestablished to monitor effectors of animal-pathogenic bacteria by quantifying the BlaM hydrolysis product [P] and fluorescence resonance energy transfer (FRET) of the substrate. The feasibility of the BlaM reporter in rice protoplasts wasevaluated by three criteria. The first criterion indicated differences between both [P] and FRET levels among wild types andOsPIP1;3-overexpressing and OsPIP1;3-silenced lines of both Nipponbare and IRBB10. The second criterion indicateddifferences between [P] and FRET levels in the presence and absence of Hpa1. The last criterion elucidated the coincidenceof PthXo1 translocation with induced expression of the PthXo1 target gene in protoplasts of Nipponbare and the coincidence of AvrXa10 translocation with induced expression of the AvrXa10 target gene in protoplasts of IRBB10. Theseresults provide an experimental avenue for real-time monitoring of bacterial T3 effector translocation into plant cells with apathological consequence.
In silico structure analysis of alphaviral RNA genomes shows diversity in the evasion of IFIT1-mediated innate immunity
(Indian Academy of Sciences, 2019-09) Barik, Sailen
The IFIT (interferon-induced proteins with tetratricopeptide repeats) family constitutes a major arm of the antiviral functionof type I interferon (IFN). Human IFIT1, the earliest discovered member of this family, inhibits several viruses of positivestrand RNA genome. IFIT1 specifically recognizes single-stranded RNA with canonical 7-methylguanylate cap at the 50 end(Cap0), and inhibits their translation by competing with eIF4E (eukaryotic initiation factor 4E), an essential factor for 50Caprecognition. Recently, a novel viral mechanism of IFIT1 suppression was reported, in which an RNA hairpin in the 50untranslated region (50UTR) of the viral genome prevented recognition by IFIT1 and enhanced virus growth. Here, I haveanalyzed the in silico predicted structures in the 50UTR of the genomes of the Alphaviruses, a large group of envelopedRNA virus with positive-sense single-stranded genome. The results uncovered a large ensemble of RNA secondarystructures of diverse size and shape in the different viruses, which showed little correspondence to the phylogeny of theviruses. Unexpectedly, the 50UTR of several viral genomes in this family did not fold into any structure, suggesting eithertheir extreme sensitivity to IFIT1 or the existence of alternative viral mechanisms of subverting IFIT1 function.
A cross-eyed geneticist’s view IV. Neurospora genes and inversions collude to cheat Mendel
(Indian Academy of Sciences, 2019-09) Kasbekar, Durgadas P
Measles virus phosphoprotein inhibits apoptosis and enhances clonogenic and migratory properties in HeLa cells
(Indian Academy of Sciences, 2019-03) Bhattacharjee, Sankhajit; Jaiswal, Rishi Kumar; Yadava, Pramod Kumar
Measles virus is the causative agent of measles, a major cause of child mortality in developing countries. Two majorproteins, coded by the viral genome, are nucleocapsid protein (N) and phosphoprotein (P). The N protein protects the viralgenomic RNA and forms ribonucleoprotein complex (RNP) together with P protein. MeV-P protein recruits the largeprotein (L), i.e. viral RNA-depended RNA polymerase (RdRp), to ensure viral replication in host cell. Apoptogenicproperties of N protein of Edmonston vaccine strain have been established in our lab previously. We investigated the role ofMeV-P protein of Edmonston vaccine strain as modulator of apoptosis in cervical cancer cell line (HeLa) and found thatMeV-P protein is anti-apoptotic and enhances cell proliferation. Measles virus is considered to be innately oncotropic virus.However, the anti-apoptotic property of MeV-P protein raises important concerns while adopting this virus as an anti-cancertherapeutic tool.
Protein profiling of SH-SY5Y neuroblastoma cells: The effect of rhein
(Indian Academy of Sciences, 2019-09) Cockova, Zuzana; Ujcikova, Hana; Telensky, Petr; Novotny, Jiri
Bone marrow mesenchymal stem cells (BMSCs) play an important role in the process of bone repair. The present studyinvestigated the effect of 5-azacytidine (AZA) and trichostatin A (TSA) on BMSC behaviors in vitro. The role of WNTfamily member 5A (WNT5A)/WNT family member 5A (WNT7A)/b-catenin signaling was also investigated. BMSCs wereisolated from a steroid-induced avascular necrosis of the femoral head (SANFH) rabbit model. The third-generation ofBMSCs was used after identification. The results revealed obvious degeneration and necrosis in the SANFH rabbit model.AZA, TSA and TSA ? AZA increased BMSC proliferation in a time-dependent fashion. AZA, TSA and TSA ? AZAinduced the cell cycle release from the G0/G1 phase and inhibited apoptosis in BMSCs. AZA, TSA and TSA ? AZAtreatment significantly decreased caspase-3 and caspase-9 activities. The treatment obviously increased the activity andrelative mRNA expression of alkaline phosphatase. The treatment also significantly up-regulated the proteins associatedwith osteogenic differentiation, including osteocalcin and runt-related transcription factor 2 (RUNX2), and Wnt/b-cateninsignal transduction pathway-related proteins b-catenin, WNT5A and WNT7A. The relative levels of Dickkopf-relatedprotein 1 (an inhibitor of the canonical Wnt pathway) decreased remarkably. Notably, TSA ? AZA treatment exhibited astronger adjustment ability than either single treatment. Collectively, the present studies suggest that AZA, TSA and TSA ?AZA promote cell proliferation and osteogenic differentiation in BMSCs, and these effects are potentially achieved via upregulation of WNT5A/WNT7A/b-catenin signaling.
MiR-212-3p inhibits cell proliferation and promotes apoptosis by targeting nuclear factor IA in bladder cancer
(Indian Academy of Sciences, 2019-09) Xiaoming, Wu; Hao, Chen; Gaoyue, Zhang; Jianhui, Wu; Wei, Zhu; Yanqin, Gu; Yi, He
Accumulating evidence suggest that microRNAs play crucial roles in the development and progression of bladder cancer(BC). Here, we found that miR-212-3p was significantly down-regulated and negatively correlated with nuclear factor IA(NFIA) in human BC tissues. Bioinformatics analysis predicted that NFIA was a target gene of miR-212-3p. Then BC celllines, T24 and J82 cells were transfected with miR-212-3p mimics or siNFIA to obtain miR-212-3p overexpression or NFIAknockdown cell lines, respectively. Quantitative real-time PCR was used to determine the expression of miR-212-3p andNFIA. Western blot analysis was utilized to detect NFIA expression. MTT assay showed either miR-212-3 overexpression orNFIA knockdown significantly inhibited the BC cell proliferation. Double staining with Annexin V-APC and 7-AAD showedthe total number of apoptotic BC cells were remarkably increased after miR-212-3p overexpression or NFIA knockdown.Collectively, our results indicated that miR-212-3p targeting NFIA might serve as a promising target for BC.

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