scholarly journals 233EXPRESSION OF INSULIN-LIKE GROWTH FACTOR (IGF) MRNA IN BOVINE CONCEPTUSES FROM EMBRYOS PRODUCED IN VIVO OR IN VITRO

2004 ◽  
Vol 16 (2) ◽  
pp. 237
Author(s):  
C.E. Farin ◽  
J.E. Alexander ◽  
K.F. Rodriguez ◽  
P.W. Farin
Keyword(s):  
Mrna Expression ◽  
Agricultural Research ◽  
Pcr Analysis ◽  
Blastocyst Development ◽  
Globin Mrna ◽  
Treatment Groups ◽  
The North ◽  
Gapdh Mrna

The objective of this study was to determine the effect of in vitro embryo production (IVP) on expression of mRNAs for IGF-1, IGF-2, IGF-1 receptor (IGF-1R), IGF-2R and GAPDH in bovine conceptuses at Day 17 of gestation. In vivo embryos (In vivo) were recovered from superovulated Holstein cows. For IVP, Holstein oocytes were matured, fertilized and then cultured in M199 with 10% serum (IVPS) or 1% BSA (IVPSR) to 72hpi. All embryos were then transferred to M199 with 10% serum and cultured to 168hpi. The same Holstein sire was used to produce all embryos. Single grade 1 blastocysts were transferred into recipient heifers. On day 10 after transfer, conceptuses were recovered, measured and stored at −80°C prior to extraction of whole cell (wc) RNA and genomic (g) DNA. wcRNA was assessed for quality based on A260/A280 ratio and visualization of 18S and 28S ribosomal RNA. Only complete (intact) conceptuses with at least 10μg high-quality wcRNA recovered after extraction were included in the analysis (In vivo, n=8; IVPS n=8; IVPSR n=7). Conceptus sex was determined by PCR analysis of gDNA. Semi-quantitative PCR assays were used to assess relative expression of all mRNAs. For each conceptus, 2μg wcRNA were mixed with 3pg rabbit globin mRNA, DNase-treated and reverse-transcribed using random hexamers. Conceptus cDNA samples (140ng each for IGF-1, IGF-2 mRNAs;; 100ng each for IGF-1R, IGF-2R, GAPDH and globin mRNAs) were assayed in duplicate within single assays. Relative mRNA expression was calculated as the ratio of band intensities representing the mRNA of interest to globin mRNA. All data were analyzed for effects of treatment (In vivo, IVPS, IVPSR or In vivo, IVP), sex, stage of blastocyst development at transfer (early, mid, expanded) and the interactions of treatment sex and treatment stage. Conceptus length (LSM±SEMmm) did not differ with treatment (In vivo: 182±55; IVPS: 257±47; IVPSR: 310±72). Expression of GAPDH mRNA was greater (P=0.02) in female conceptuses (1.31±0.11) than in males (0.96±0.07) and did not differ (P=0.14) between treatment groups (In vivo: 1.30±0.11, IVPS: 1.00±0.09, IVPSR: 1.10±0.14). However, when the IVP groups were combined and compared to In vivo controls, expression of GAPDH mRNA was both greater (P=0.002) in females (1.38±0.10) than in males (0.94±0.06) and differed (P=0.03) with treatment (In vivo: 1.30±0.10, IVP: 1.01±0.07). Expression of IGF-1 and IGF-2 mRNA was not detected in the majority of conceptuses. Expression of IGF-1R mRNA was greater (P=0.02) in female (1.19±0.10) than in male conceptuses (0.87±0.07) and differed (P=0.02) with stage of blastocyst development at transfer (early: 0.79±0.07; mid: 1.23±0.13; expanded: 1.07±0.11). There was no effect of treatment on expression of IGF-1R mRNA. Expression of IGF-2R mRNA differed (P=0.03) with treatment (In vivo: 0.85±0.10; IVPS: 0.76±0.09; IVPSR: 0.33±0.13). In conclusion, expression of both imprinted and non-imprinted genes is altered in conceptuses resulting from embryos produced in vitro. Supported by the North Carolina Agricultural Research Service.

Healing effects of curcumin nanoparticles in deep tissue injury mouse model.

2020 ◽  
Vol 18 ◽  
Author(s):  
Zirui Zhang ◽  
Shangcong Han ◽  
Panpan Liu ◽  
Xu Yang ◽  
Jing Han ◽  
...  
Keyword(s):  
Wound Healing ◽  
Mrna Expression ◽  
Tissue Injury ◽  
Inflammatory Cells ◽  
Pcr Analysis ◽  
Protective Effects ◽  
Deep Tissue ◽  
Deep Tissue Injury

Background: Chronic inflammation and lack of angiogenesis are the important pathological mechanisms in deep tissue injury (DTI). Curcumin is a well-known anti-inflammatory and antioxidant agent. However, curcumin is unstable under acidic and alkaline conditions, and can be rapidly metabolized and excreted in the bile, which shortens its bioactivity and efficacy. Objective: This study aimed to prepare curcumin-loaded poly (lactic-co-glycolic acid) nanoparticles (CPNPs) and to elucidate the protective effects and underlying mechanisms of wound healing in DTI models. Methods: CPNPs were evaluated for particle size, biocompatibility, in vitro drug release and their effect on in vivo wound healing. Results : The results of in vivo wound closure analysis revealed that CPNP treatments significantly improved wound contraction rates (p<0.01) at a faster rate than other three treatment groups. H&E staining revealed that CPNP treatments resulted in complete epithelialization and thick granulation tissue formation, whereas control groups resulted in a lack of compact epithelialization and persistence of inflammatory cells within the wound sites. Quantitative real-time PCR analysis showed that treatment with CPNPs suppressed IL-6 and TNF-α mRNA expression, and up-regulated TGF-β, VEGF-A and IL-10 mRNA expression. Western blot analysis showed up-regulated protein expression of TGF-β, VEGF-A and phosphorylatedSTAT3. Conclusion: Our results showed that CPNPs enhanced wound healing in DTI models, through modulation of the JAK2/STAT3 signalling pathway and subsequent upregulation of pro-healing factors.

1 ARGININE SUPPLEMENTATION IN VITRO INCREASES PORCINE EMBRYO DEVELOPMENT AND AFFECTSmRNA TRANSCRIPT EXPRESSION

2011 ◽  
Vol 23 (1) ◽  
pp. 107 ◽  
Author(s):  
B. K. Bauer ◽  
L. D. Spate ◽  
C. N. Murphy ◽  
R. S. Prather
Keyword(s):  
Treatment Group ◽  
Mrna Expression ◽  
Embryo Development ◽  
Blastocyst Stage ◽  
Sequencing Analysis ◽  
Treatment Groups ◽  
Porcine Embryo ◽  
Arginine Supplementation

In vitro culture systems are suboptimal as compared to in vivo. Previous next-generation sequencing analysis of in vivo fertilized and in vitro cultured (IVC) or in vivo cultured (IVV) porcine blastocyst stage embryos identified an arginine transporter (SLC7A1) expressed 63 fold higher in IVC compared to IVV blastocysts (Bauer et al. 2010 Biol. Reprod. Epub ahead of print). Arginine catabolism may play important roles in placental and conceptus growth and development as it is a substrate for synthesis of nitric oxide synthase and polyamines. The objective of this study was to determine the effects arginine had on both embryo development and mRNA expression in in vitro fertilized embryos. Cumulus–oocyte complexes were matured for 44 h in M199 supplemented with EGF, FSH, and LH. Oocytes with a visible polar body (metaphase II) were selected and fertilized in modified Tris Buffered Medium for 5 h and then placed into one of 5 treatment groups (Porcine Zygote Medium 3 (PZM3) with 0 mM, 0.12 mM (current concentration of arginine in PZM3), 0.36 mM, 0.72 mM, or 1.69 mM arginine). Twenty-eight hours post-fertilization, cleaved embryos were selected and moved into 25 μL drops of respective culture media and cultured to day 6 in 5% CO2, 5% O2, 90% N2 at 38.5°C. To determine the effect arginine had on development the percent of embryos that made it to the blastocyst stage for each treatment group were analysed using PROC GLM in SAS (SAS Institute, Cary, NC). A least significant difference post test comparison was completed to determine if significant differences existed between treatment groups (a,b,cP < 0.05). The percentage of cleaved embryos on Day 6 that developed to blastocyst was 57.2%b,c, 50.2%c, 67.3%a,b, 67.3%a,b, 70.4%a (N = 147, 163, 150, 120, and 134) in 0 mM, 0.12 mM, 0.36 mM, 0.72 mM, and 1.69 mM arginine, respectively. Real-time PCR was then completed to assess the affect arginine supplementation had on SLC7A1 mRNA expression. Three biological replicates, each containing 10 blastocyst pools to ensure enough starting material, were collected for each treatment group. RNA was isolated from each sample and 5 μL was linearly amplified (NuGEN Ovation Pico WTA System) so multiple genes could be compared and then purified using Bio-Rad MicroSpin Columns. Expression levels were calculated relative to the reference sample and the housekeeping gene, YWHAG. The ΔΔCT values were log-transformed and analysed using PROC GLM in SAS. The expression of SLC7A1 mRNA was decreased (P = 0.0006) compared to PZM3 in the 1.69 mM arginine group. These results illustrate the positive effects that additional arginine may be having on porcine embryo development during culture from the 2-cell to the blastocyst stage. Supplementing arginine to a final concentration of 1.69 mM during culture increases development of porcine embryos to blastocyst compared to PZM3 and also decreases the expression of SLC7A1. Evaluation of the transcriptional profile appears to be a good method of letting the embryo tell us what it needs for development, and in this case arginine. Funded by F21C.

scholarly journals 150 ENVIRONMENT OF THE EARLY EMBRYO AND ITS EFFECT ON DEVELOPMENT AND POSTNATAL LIFE

2005 ◽  
Vol 17 (2) ◽  
pp. 225
Author(s):  
A. Watkins ◽  
A. Wilkins ◽  
T. Papenbrock ◽  
C. Osmond ◽  
M. Hanson ◽  
...  
Keyword(s):  
Blood Pressure ◽  
Systolic Blood Pressure ◽  
Embryo Transfer ◽  
Litter Size ◽  
Early Embryo ◽  
Blastocyst Development ◽  
Treatment Groups ◽  
Elevated Systolic Blood Pressure

We have investigated the impact of mouse early embryo in vitro culture environment on (a) short-term blastocyst development and (b) long-term postnatal growth and physiology after embryo transfer. In vitro-developed blastocysts, cultured from the 2-cell stage, had reduced inner cell mass (ICM) and trophectoderm (TE) cell numbers when compared to in vivo-derived blastocysts at 96 h post-hCG (n = 13–39, P < 0.05). Despite the retardation in blastocyst development, the ICM:TE ratio was equivalent in both treatment groups. Using embryo transfer techniques, we compared the postnatal development of embryos cultured in vitro from the 2-cell to the blastocyst stage (termed “in vitro” mice) with offspring generated from blastocysts developed in vivo, but which also underwent embryo transfer (termed “in vivo” mice). These two treatment groups were in turn compared with mice derived from naturally mated mothers, which had their mean litter size at birth adjusted to a size comparable with that of the in vitro and in vivo mice (a mean of 6 animals) and which had not been transferred. All data were analyzed using a multilevel random effects regression model which took into account between-mother and within-mother variation in litter size for parameters measured from individual animals. No significant differences in birth weight were observed between in vitro and in vivo offspring. However, in vitro offspring were significantly lighter than in vivo offspring in a gender-dependent manner at 2 weeks of age (males, P = 0.009) and at 6 and 11 weeks of age (females, P = 0.037 and 0.035, respectively). In addition, at 4 weeks of age, the in vivo males became significantly lighter when compared to the naturally mated males (P = 0.034). At 8 weeks of age, the in vivo females had a significantly elevated systolic blood pressure when compared to the in vitro females (P = 0.003); however, at 21 weeks of age, both in vitro males and females had a significantly elevated blood pressure when compared to in vivo offspring (P < 0.003). At 8, 15, and 21 weeks of age, offspring derived from transferred embryos developed with significantly elevated systolic blood pressure when compared to non-embryo transfer offspring (P < 0.05). No significant differences in serum angiotensin-converting enzyme activity (a potent regulator of systolic blood pressure) was observed between the treatment groups. Significantly altered liver:body weight ratios were observed between the in vitro and in vivo males, and between the in vitro and the naturally mated (6) females (P < 0.038). All of the above data are independent of litter size. These data support the hypothesis that early embryo environment can influence postnatal growth and cardiovascular physiology. This research was funded by an MRC research grant to TPF, and by a DOHaD studentship.

scholarly journals Birth of lambs of a pre-determined sex after in vitro production of embryos using frozen–thawed sex-sorted and re-frozen–thawed ram spermatozoa

Reproduction ◽  
2004 ◽  
Vol 127 (5) ◽  
pp. 557-568 ◽  
Author(s):  
F K Hollinshead ◽  
G Evans ◽  
K M Evans ◽  
S L Catt ◽  
W M C Maxwell ◽  
...  
Keyword(s):  
High Speed ◽  
Gradient Centrifugation ◽  
In Vitro Production ◽  
Blastocyst Development ◽  
Embryo Survival ◽  
Treatment Groups ◽  
Sperm Migration ◽  
And Control

The characteristics and functional capacity of ram spermatozoa frozen–thawed prior to and after flow cytometric sorting was assessed after incubation (37 °C; 6 h),in vitrofertilisation (IVF), and transfer of fresh and vitrifiedin vitroproduced embryos. Frozen-thawed spermatozoa from two rams were allocated to four treatment groups: (i) non-sorted (Control); (ii) sorted (FS); (iii) sorted then re-frozen (FSF) and (iv) re-frozen control (FCF). Frozen-thawed samples were separated into X- and Y-chromosome bearing spermatozoa using a high-speed sperm sorter after density gradient centrifugation (X: 88 ± 1.5% and Y: 87 ± 1.1% purity). After 6 h incubation (37 °C), the percentage of motile spermatozoa was higher (P< 0.001) for FS (84 ± 2.0%) compared with all other treatments (Control: 36 ± 3.3%, FSF: 28 ± 3.1%, FCF: 20 ± 2.0%). In a sperm migration test greater numbers of FS spermatozoa penetrated 5 mm into the artificial cervical mucus compared with spermatozoa from all other treatments (152 ± 39.4 vs 31 ± 9.2 spermatozoa respectively;P< 0.05). Fertilisation and cleavage rates were higher (P< 0.05) forin vitromatured oocytes inseminated with Control compared with FSF spermatozoa. However, the Day 7 blastocyst development rate was higher for oocytes inseminated with FSF (62.2%) than FS and Control spermatozoa (52.7 and 50.0%;P< 0.05). The number of ewes pregnant (Day 60), lambing and thein vivoembryo survival rate was greater (P< 0.01) after the transfer of fresh embryos rather than vitrified embryos derived from X- and Y-spermatozoa (67.6, 64.7 and 41.2% vs 29.6, 25.9 and 14.8% respectively). Twenty-six of the 30 (86.7%) lambs derived from sex-sorted spermatozoa were of the correct sex. These results demonstrate that frozen–thawed ram spermatozoa can be sex-sorted for immediate or future use after re-cryopreservation and, in conjunction with IVF and embryo transfer, can be used to efficiently produce offspring of pre-determined sex.

scholarly journals Normal gonadotropin production and fertility in gonadotrope-specific Bmpr1a knockout mice

2016 ◽  
Vol 229 (3) ◽  
pp. 331-341 ◽  
Author(s):  
Xiang Zhou ◽  
Ying Wang ◽  
Luisina Ongaro ◽  
Ulrich Boehm ◽  
Vesa Kaartinen ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Knockout Mice ◽  
Reproductive Organ ◽  
Normal Serum ◽  
Receptor Protein ◽  
Pcr Analysis ◽  
Type I ◽  
Pituitary Cells

Pituitary follicle-stimulating hormone (FSH) synthesis is regulated by transforming growth factorβsuperfamily ligands, most notably the activins and inhibins. Bone morphogenetic proteins (BMPs) also regulate FSHβ subunit (Fshb) expression in immortalized murine gonadotrope-like LβT2 cells and in primary murine or ovine primary pituitary cultures. BMP2 signals preferentially via the BMP type I receptor, BMPR1A, to stimulate murine Fshb transcription in vitro. Here, we used a Cre–lox approach to assess BMPR1A’s role in FSH synthesis in mice in vivo. Gonadotrope-specific Bmpr1a knockout animals developed normally and had reproductive organ weights comparable with those of controls. Knockouts were fertile, with normal serum gonadotropins and pituitary gonadotropin subunit mRNA expression. Cre-mediated recombination of the floxed Bmpr1a allele was efficient and specific, as indicated by PCR analysis of diverse tissues and isolated gonadotrope cells. Furthermore, BMP2 stimulation of inhibitor of DNA binding 3 expression was impaired in gonadotropes isolated from Bmpr1a knockout mice, confirming the loss of functional receptor protein in these cells. Treatment of purified gonadotropes with small-molecule inhibitors of BMPR1A (and the related receptors BMPR1B and ACVR1) suppressed Fshb mRNA expression, suggesting that an autocrine BMP-like molecule might regulate FSH synthesis. However, deletion of Bmpr1a and Acvr1 in cultured pituitary cells did not alter Fshb expression, indicating that the inhibitors had off-target effects. In sum, BMPs or related ligands acting via BMPR1A or ACVR1 are unlikely to play direct physiological roles in FSH synthesis by murine gonadotrope cells.

scholarly journals In vitro evaluation of chitosan-DNA plasmid complex encoding Jembrana disease virus Env-TM protein as a vaccine candidate

2019 ◽  
Vol 63 (1) ◽  
pp. 7-16 ◽  
Author(s):  
Januar Ishak ◽  
Lalu Unsunnidhal ◽  
Ronny Martien ◽  
Asmarani Kusumawati
Keyword(s):  
Mrna Expression ◽  
Dna Vaccine ◽  
Hela Cells ◽  
Restriction Analysis ◽  
Initial Study ◽  
Pcr Analysis ◽  
Colony Pcr ◽  
Dna Complex

AbstractIntroduction:The development of Jembrana disease vaccine is an important effort to prevent losses in the Bali cattle industry in Indonesia. This study aims to prepare a Jembrana DNA vaccine encoding the transmembrane portion of the envelope protein in pEGFP-C1 and test the success of its delivery in culture cells using a chitosan-DNA complex.Material and Methods:Cloning of the DNA vaccine was successfully performed onE. coliDH5α and confirmed by colony PCR, restriction analysis and sequencing. The plasmids were prepared as a chitosan complex using the complex coacervation method and physicochemically characterised using a particle size analyser. A transfection assay was performed in HeLa cells with 4 h exposure, and mRNA expression was assessed at 24 h post transfection.Results:With a 1:2 (wt./wt.) ratio of DNA and chitosan, the complexes have a mean diameter of 236 nm, zeta potential value of + 17.9 mV, and showed no high toxicity potential in the HeLa cells. This complex successfully delivered the DNA into cells, as shown by the presence of a specific RT-PCR product (336 bp). However, the real-time PCR analysis showed that the delivery with chitosan complex resulted in lower target mRNA expression when compared with a commercial transfecting agent.Conclusion:pEGFP-env-tm JDV as a candidate vaccine can be delivered as the chitosan-DNA complex and be expressed at the transcription levelin vitro. This initial study will be used for further improvement and evaluationin vivo.

Comparison of two approaches to nuclear transfer in the bovine: hand-made cloning with modifications and the conventional nuclear transfer technique

2005 ◽  
Vol 17 (5) ◽  
pp. 573 ◽  
Author(s):  
R. Tayfur Tecirlioglu ◽  
Melissa A. Cooney ◽  
Ian M. Lewis ◽  
Natasha A. Korfiatis ◽  
Renee Hodgson ◽  
...  
Keyword(s):  
Cell Lines ◽  
Nuclear Transfer ◽  
Cell Number ◽  
Developmental Competence ◽  
Blastocyst Development ◽  
Total Cell Number ◽  
Treatment Groups ◽  
Fusion Efficiency

The aim of the present study was to compare the in vitro and in vivo developmental competence of hand-made cloning (HMC) embryos with the conventional nuclear transfer (NT) method using five somatic cell lines and in vitro-fertilised (IVF; control) embryos. Modifications to the HMC procedure included fusion efficiency optimisation, effect of cytoplasmic volume and cloned embryo aggregation. The developmental competence of blastocysts from each of the treatment groups and cell lines used was assessed following transfer to 345 recipients. Vitrification was also used to enable management of recipient resources and to assess the susceptibility of membranes to cryopreservation following zona removal. Increasing cytoplasmic volume to 150% or aggregating two embryos improved the blastocyst development rate and increased the total cell number. Although HMC embryo transfers established a significantly higher pregnancy rate on Day 30 than fresh IVF or NT embryo transfers, the overall outcome in terms of cloned live births derived from either fresh or vitrified/thawed HMC or NT embryo transfers across the five cell lines did not differ. The birth and continued survival of clones produced with HMC technology with equivalent efficiency to NT shows that it can be used as an alternative method for the generation of cloned offspring in the bovine.

scholarly journals Efficacy of ruxolitinib as inducer of fetal hemoglobin in primary erythroid cultures from sickle cell and beta-thalassemia patients

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Alice Pecoraro ◽  
Antonio Troia ◽  
Aurelio Maggio ◽  
Rosalba Di Marzo
Keyword(s):  
Mrna Expression ◽  
Sickle Cell ◽  
Cultured Cells ◽  
Globin Gene ◽  
Beta Thalassemia ◽  
Erythroid Progenitors ◽  
Globin Mrna ◽  
Gamma Globin

High levels of HbF may ameliorate the clinical course of β-thalassaemia and SCD. Hydroxyurea (HU) is the only HbF inducer approved for the treatment of patients. However not all patients respond to the treatment, for this reason it is noteworthy to identify new HbF inducers. Ruxolitinib is a JAK inhibitor that decreases the phosphorilation of STAT proteins. In particular STAT3 is a repressor of gamma-globin gene. The decrease of STAT3 phosphorilation could derepress gamma-globin gene and reactivate its trascription. In this study we evaluated the efficacy of ruxolitinib as inducer of HbF production. The analyses were performed in cultured erythroid progenitors from 16 beta-thalassemia intermedia (TI) and 4 sickle cell disease (SCD) patients. The use of quantitative RT-PCR technique allowed us to determine the increase of gamma-globin mRNA expression in human erythroid cultured cells treated with ruxolitinib. The results of our study demonstrated an increase in vitro of gamma-globin mRNA expression in almost all patients. These data suggest that ruxolitinib could be a good candidate to be used in vivo for the treatment of hemoglobinopathies.

scholarly journals Filociclovir Is an Active Antiviral Agent against Ocular Adenovirus Isolates In Vitro and in the Ad5/NZW Rabbit Ocular Model

2021 ◽  
Vol 14 (4) ◽  
pp. 294
Author(s):  
Eric G. Romanowski ◽  
Islam T. M. Hussein ◽  
Steven C. Cardinale ◽  
Michelle M. Butler ◽  
Lucas R. Morin ◽  
...  
Keyword(s):  
Antiviral Activity ◽  
Topical Treatment ◽  
Antiviral Agent ◽  
Human Adenovirus ◽  
Treatment Groups ◽  
Ocular Infections ◽  
The Mean ◽  
Eye Infection

Presently, there is no FDA- or EMA-approved antiviral for the treatment of human adenovirus (HAdV) ocular infections. This study determined the antiviral activity of filociclovir (FCV) against ocular HAdV isolates in vitro and in the Ad5/NZW rabbit ocular model. The 50% effective concentrations (EC50) of FCV and cidofovir (CDV) were determined for several ocular HAdV types using standard plaque reduction assays. Rabbits were topically inoculated in both eyes with HAdV5. On day 1, the rabbits were divided into four topical treatment groups: (1) 0.5% FCV 4x/day × 10 d; (2) 0.1% FCV 4x/day × 10 d; (3) 0.5% CDV 2x/day × 7 d; (4) vehicle 4x/day × 10 d. Eyes were cultured for virus on days 0, 1, 3, 4, 5, 7, 9, 11, and 14. The resulting viral eye titers were determined using standard plaque assays. The mean in vitro EC50 for FCV against tested HAdV types ranged from 0.50 to 4.68 µM, whereas those treated with CDV ranged from 0.49 to 30.3 µM. In vivo, compared to vehicle, 0.5% FCV, 0.1% FCV, and 0.5% CDV produced lower eye titers, fewer numbers of positive eye cultures, and shorter durations of eye infection. FCV demonstrated anti-adenovirus activity in vitro and in vivo.

scholarly journals Targeting a Lipid Desaturation Enzyme, SCD1, Selectively Eliminates Colon Cancer Stem Cells through the Suppression of Wnt and NOTCH Signaling

Cells ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 106
Author(s):  
Yeongji Yu ◽  
Hyejin Kim ◽  
SeokGyeong Choi ◽  
JinSuh Yu ◽  
Joo Yeon Lee ◽  
...  
Keyword(s):  
Colon Cancer ◽  
Notch Signaling ◽  
Cultured Cells ◽  
Pcr Analysis ◽  
Marker Genes ◽  
Dependent Manner ◽  
Lipid Desaturation ◽  
Novel Target

The elimination of the cancer stem cell (CSC) population may be required to achieve better outcomes of cancer therapy. We evaluated stearoyl-CoA desaturase 1 (SCD1) as a novel target for CSC-selective elimination in colon cancer. CSCs expressed more SCD1 than bulk cultured cells (BCCs), and blocking SCD1 expression or function revealed an essential role for SCD1 in the survival of CSCs, but not BCCs. The CSC potential selectively decreased after treatment with the SCD1 inhibitor in vitro and in vivo. The CSC-selective suppression was mediated through the induction of apoptosis. The mechanism leading to selective CSC death was investigated by performing a quantitative RT-PCR analysis of 14 CSC-specific signaling and marker genes after 24 and 48 h of treatment with two concentrations of an inhibitor. The decrease in the expression of Notch1 and AXIN2 preceded changes in the expression of all other genes, at 24 h of treatment in a dose-dependent manner, followed by the downregulation of most Wnt- and NOTCH-signaling genes. Collectively, we showed that not only Wnt but also NOTCH signaling is a primary target of suppression by SCD1 inhibition in CSCs, suggesting the possibility of targeting SCD1 against colon cancer in clinical settings.

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