ABSTRACT
ABSTRACT Introduction: Acute promyelocytic leukemia currently presents an excellent chance of cure with protocols based on all-trans-retinoic acid (ATRA) and anthracycline or only differentiation agents. However, high early mortality rates continue to be reported Methods: Between 2000 and 2018, patients were enrolled and retrospectively analyzed by medical records. A modified AIDA protocol, with a 1-year shortening of the treatment duration, reduction in the number of drugs and a strategy to reduce early mortality by the postponement of the initiation of anthracyclines were employed. Overall and event-free survival rates and toxicity were analyzed Results: Thirty-two patients were enrolled, of whom 56% were female, with a median age of 12 years and 34% belonged to the high-risk group. Two patients had the hypogranular variant and three had another cytogenetic alteration, in addition to the t(15;17). The median start of the first anthracycline dose was 7 days. There were two early deaths (6%) due to central nervous system (CNS) bleeding. All patients achieved molecular remission after the consolidation phase. Two children relapsed and were rescued by arsenic trioxide and hematopoietic stem cell transplantation. The presence of disseminated intravascular coagulation (DIC) at diagnosis (p = 0.03) was the only factor with survival impact. The five-year event-free survival (EFS) was 84% and 5-year overall survival (OS) was 90% Conclusion: The survival results were comparable to those found in the AIDA protocol, with a low rate of early mortality in relation to the Brazilian reality.
ABSTRACT
Objective:To observe the effect of the antioxidant N-acetylcysteine (NAC) and selective endoplasmic reticulum stress response inhibitor salubrinal on the apoptosis of retinal pigment epithelial cells induced by all-trans-retinoic acid (ATRA).Methods:Human ARPE-19 cell line was used as the experimental cell line, and was divided into normal control group cultured with complete medium, model control group cultured with complete medium containing 10 μmol/L ATRA, NAC treatment group cultured with complete medium containing 10 μmol/L ATRA+ 5 mmol/L NAC, salubrinal group cultured with complete medium containing 10 μmol/L ATRA+ 40 μmol/L salubrinal, NAC+ salubrinal group cultured with complete medium containing 10 μmol/L ATRA+ 5 mmol/L NAC+ 40 μmol/L salubrinal.After 24-hour culture, apoptosis rate, multicaspase level and reactive oxygen species (ROS) level of ARPE-19 cells were detected by flow cytometry.The expressions of vascular endothelial growth factor A (VEGF-A), C/EBP-homologous protein (CHOP), cleaved-caspase 3 in cells were detected by Western blot.Results:There were significant differences in the apoptosis rate, multicaspase and ROS levels among the five groups ( F=113.23, 602.41, 160.39; all at P<0.001). The apoptosis rate, multicaspase and ROS levels of normal control group, NAC treatment group, salubrinal group and NAC+ salubrinal group were significantly lower than those of model control group (all at P<0.05). There were significant differences in the expression levels of VEGF-A, CHOP and cleaved-caspase 3 among the five groups ( F=24.62, 36.35, 60.25; all at P<0.001). The protein expression levels of VEGF-A, CHOP and cleaved-caspase 3 of normal control group, NAC treatment group, salubrinal group and NAC+ salubrinal group were significantly lower than those of model control group (all at P<0.05). Conclusions:ATRA can induce RPE cells to produce oxidative stress and endoplasmic reticulum stress injury, which leads to apoptosis.NAC and salubrinal can effectively reduce the RPE cell apoptosis by inhibiting stress response.
ABSTRACT
Objective@# To explore the preventive effect of nicotinamide (NAM) on cleft palate induced by all-trans retinoic acid (RA), to provide research evidence for the prevention of cleft palate. @*Methods @#The mouse cleft palate model was induced by intragastric administration of 70 mg/kg all-trans retinoic acid at embryonic day 10.5 (E10.5) in the control group. The mouse cleft palate model was treated by caudal vein injection of 20 mg/kg NAM at E8.5 to E13.5 in the experimental group (1). The cleft palate model was treated by caudal vein injection of 40 mg/kg NAM at E8.5-E13.5 in the experimental group (2). The cleft palate of fetal rats was observed by laparotomy on E16.5 and statistically analyzed. Annexin V-FITC/PI double staining was used to detect the apoptosis of mouse embryonic palatal mesenchyme (MEPM) cells treated with RA 1 μmol/L (RA 1 group), NAM 200 μmol/L (NAM 200 group), and both NAM 200 μmol/L and RA 1 μmol/L (NAM 200+RA 1 group) for 24 hours by flow cytometry and the apoptosis rate in groups were compared. Culture without RA or NAM was used as a control. @*Results @# The cleft palate rate in the control group was 98%. The cleft palate rate in experimental group (1) was 87%. There was no significant difference between groups (P>0.05). The cleft palate rate in the experimental group (2) was 63%, compared with the control group, there was a significant difference (P<0.01). The cell apoptosis rate was 16.53%±2.89% in the CONTROL group. The cell apoptosis rate was 22.9%±1.85% in the RA 1 group, which was a significant increase compared with the CONTROL group (P<0.01). The apoptotic rate of the NAM 200 group was 9.23%±1.39%, which was a significant decrease compared with NA 1 group (P<0.01). The apoptosis rate of the NAM 200+RA 1 group was 14.9%±7.67%, which was a significant decrease compared with the RA 1 group (P<0.01).@*Conclusion@#NAM can prevent cleft palate. 40 mg/kg nicotinamide during pregnancy is an effective concentration for the prevention of RA-induced cleft palate. The mechanism by which NAM prevents cleft palate may be that NAM inhibits RA-induced apoptosis of MEPM cells.
ABSTRACT
Objective: To evaluate the safety and efficacy of Compound Huangdai Tablets (Realgar-Indigo Naturalis formula, RIF) combined with all-trans retinoic acid (ATRA) to treat acute promyelocytic leukemia (APL). Methods: This study was registered in PROSPERO (CRD42018108118). The relevant literatures on RIF treatment of APL were systematically searched in the following databases: China National Knowledge Infrastructure, Wanfang, VIP Medical Information System, Chinese Biomedical Database, EMBASE, Cochrane Library, and PubMed. The quality of the included studies was evaluated and Review Manager 5.3 software and Stata 13.0 software were used to perform the Meta-analysis. In addition, this study used the method of network pharmacology to conduct a preliminary exploration of the mechanism of RIF on APL. Results: The study included 12 studies involving 775 APL patients. The Meta-analysis showed that there was no significant difference (P 0.05) between the RIF group and the arsenic trioxide (ATO) group for primary outcomes, secondary outcomes apart from liver dysfunction. The incidence of liver dysfunction (P = 0.006) in the RIF group were significantly lower than those in the ATO group. In addition, the cost of maintenance therapy in the RIF group was significantly lower (P 0.05) than the ATO group. Besides, the active ingredients in RIF mainly act on targets proteins such as ACHE, NCOA2, RXRA, and then play a role in the treatment of APL through regulating multiple molecular mechanisms, such as TP53 regulates transcription of cell cycle genes, nuclear receptor transcription pathway. Conclusion: There was no significant difference in efficacy of oral RIF combined with ATRA compared with intravenous ATO combined with ATRA for the treatment of APL. The oral RIF exposed patients to less risk, offered more convenience and had lower prices. RIF can treat APL by multi-target and multi-pathway interventions that inducing apoptosis of APL cells and inhibiting the proliferation of APL cells, and so on. Therefore, oral RIF in the treatment of APL is worthy of further research and development.
ABSTRACT
OBJECTIVE@#To investigate the effects of decitabine (DEC) combined with all-trans retinoic acid (ATRA) on the number of immune cells, efficacy and adverse reactions in the treatment of myeloid neoplasms patients.@*METHODS@#Eighty-four patients with myeloid tumors, including AML, MDS-EB-1 or MDS-EB-2 treated by the regimen containing decitabine in our hospital from January 2009 to October 2019 were enrolled and retrospectively analyzed, among the patients, 21 patients treated with DEC alone, 24 patients treated with DEC combined with ATRA (DEC/ATRA) and 39 patients treated with DEC combined with G-CSF priming regimen (DEC/priming). The changes of peripheral blood immune cell levels before and after treatment of the patients between the three groups were compared, and the differences in clinical efficacy and adverse reactions of the patients between the three groups were also compared.@*RESULTS@#There was no statistical differences in the number of immune cells among the patients in the three groups before treatment (P>0.05). NK cell levels decreased significantly in the patients in DEC and DEC/ATRA group after treatment (P<0.05); After treatment, the levels of CD8+ and CD3+T cells in the patients treated by DEC /priming regimen significantly increased (P<0.05), while the levels of CD3-HLA-DR+ B cells significantly decreased (P<0.05). The overall response rate (ORR) of the patients in DEC/ATRA group (75%) and DEC/priming group (74.36%) was significantly higher than 42.86% in DEC monotherapy group, and the differences showed statistically significant (P<0.05), while the ORR between the patients in DEC/ATRA and DEC/priming group showed no statistic differences (P>0.05). There were no statistical differences in overall survival (OS) and incidence of bleeding between the patients in the three groups (P>0.05). The incidences of grade 3 to 4 bone marrow suppression and the infection rate of the patients in DEC monotherapy and DEC/ATRA group were significantly lower than that in DEC/priming regimen group after treatment (all P<0.05), however, there was no statistical difference between DEC monotherapy and the DEC/ATRA group.@*CONCLUSION@#The efficacy of DEC/ATRA on myeloid neoplasms is comparable to that of DEC/priming regimen, and the anti-myeloid tumor effect of DEC/ATRA regimen may be related to the regulation of NK cells and T cells.
Subject(s)
Humans , Antineoplastic Combined Chemotherapy Protocols , Decitabine/therapeutic use , Granulocyte Colony-Stimulating Factor/therapeutic use , Leukemia, Myeloid, Acute/drug therapy , Retrospective Studies , Treatment Outcome , Tretinoin/therapeutic useABSTRACT
ABSTRACT Introduction: We performed cost-effectiveness and cost-utility analyses of the modified International Consortium on Acute Promyelocytic Leukemia protocol in Mexico for the treatment of acute promyelocytic leukemia Acute Promyelocytic Leukemia. Methods: We performed a three-state Markov analysis: stable disease (first line complete response [CR]), disease event (relapse, second line response and CR) and death. The modified IC-APL protocol is composed of three phases: induction, consolidation and maintenance. Cost and outcomes were used to calculate incremental cost-effectiveness ratios (ICERs); quality-adjusted life-years were used to calculate incremental cost-utility ratios (ICURs). Results: The CR was achieved in 18 patients (90%), treated with the IC-APL protocol as the first-line option; one patient (5%) died in induction, another one never achieved CR (5%); of the 18 patients that achieved CR, 1 relapsed (5.5%). The median treatment cost of the IC-APL protocol was $21,523 USD. The average life-year in our study was 7.8 years, while the average quality-adjusted life-year (QALY) was 6.1 years. When comparing the ICER between the IC-APL and the all-trans retinoic acid (ATRA) plus arsenic trioxide (ATO) protocols, we found the different costs of $6497, $19,133 and $17,123 USD in Italy, the USA and Canada, respectively. In relation to the ICUR, we found the different costs to be $13,955 and $11,979 USD in the USA and Canada, respectively. Conclusion: Taking into account the similar response rates, lower cost and easy access to the modified IC-APL regimen, we consider it a cost-effective and cost-utility protocol, deeming it the treatment of choice for our population.
Subject(s)
Humans , Male , Female , Adolescent , Adult , Middle Aged , Aged , Leukemia, Promyelocytic, Acute/diagnosis , Leukemia, Promyelocytic, Acute/drug therapy , Tretinoin/therapeutic use , Clinical Protocols , Cost-Benefit AnalysisABSTRACT
Objective:To investigate the clinical efficacy of compound Huangdai tablets combined with all-trans retinoic acid in the treatment of acute promyelocytic leukemia. Methods:A total of 120 patients with acute promyelocytic leukemia who received treatment in the First People's Hospital of Huzhou, China between February 2013 and February 2015 were included in this study. They were randomly assigned to receive either all-trans retinoic acid, mercaptopurine and methotrexate in combination (control group, n = 60) or all-trans retinoic acid combined with compound Huangdai tablets (study group, n = 60). The time to hemoglobin rebounding, the time to platelet count rebounding, and the time to first complete remission were recorded. The recurrence and death of patients during 5 years of follow up were recorded. Adverse drug reactions during the treatment period were recorded. Results:The time to hemoglobin rebounding, the time to platelet count rebounding, and the time to first complete remission in the study group were (19.56 ± 2.61) days, (20.42 ± 2.73) days, (1.74 ± 0.45) months, respectively, which were significantly shorter than those in the control group [(28.42 ± 3.85) days, (30.63 ± 4.02) days, (3.31 ± 0.69) months, t = 10.328, 9.746, 8.521, P < 0.001]. The 1-year and 5-year recurrence rate in the study group were 11.37% and 21.67% respectively, which were significantly lower than those in the control group [28.33%, 41.67%, χ2 = 5.208, 5.546, P = 0.022, 0.019]. 5-year mortality rate in the study group was significantly lower than that in the control group [8.33% vs. 25.00%, χ2 = 6.000, P = 0.014]. There was no significant difference in adverse drug reaction between study and control group [28.33% vs. 26.67%, χ2 = 0.042, P = 0.838]. Conclusion:Compound Huangdai tablets combined with all-trans retinoic acid can effectively shorten the remission time of acute promyelocytic leukemia, reduces the disease recurrence rate and mortality rate, and is highly safe.
ABSTRACT
Objective:To investigate all-trans retinoic acid(ATRA)-induced apoptosis signaling pathway in ARPE-19 cells in vitro. Methods:The APRE-19 cell was treated with different concentrations of ATRA for 24 hours and 48 hours.Cell counting kit-8 (CCK-8) was used to detect the cell viability in order to determine the experimental concentration range.Flow cytometry and Western blot method were performed to evaluate the apoptosis and caspase related protein levels in ARPE-19 cells treated with 0, 2.5, 5, 10, 15 and 20 μmol/L of ATRA for 24 hours.Flow cytometry was used to detect the reactive oxygen species (ROS) and multicaspase levels and quantitative real-time PCR was carried out to determine the mRNA relative expression levels of caspase related proteins in ARPE-19 cells treated with 0, 2.5, 5, 10 and 20 μmol/L of ATRA, and 0 μmol/L ATRA group was used as the blank control group.Results:CCK-8 test showed that the half maximal inhibitory concentration of ARPE-19 cells treated with different concentrations of ATRA for 24 hours and 48 hours were 13.88 μmol/L and 11.99 μmol/L, respectively.The cell survival rates of ARPE-19 cells treated with different concentrations of ATRA for 24 hours and 48 hours were significantly different ( F=176.60, 350.30; both at P<0.01). When cultured for 24 hours, the cell survival rates of ARPE-19 cells in the 2 μmol/L and 6 μmol/L of ATRA groups were higher than that of the blank control group (both at P<0.05), and the cell survival rates of ARPE-19 cells in the 12, 14, 16, 18 and 20 μmol/L of ATRA groups were lower than that of the blank control group (all at P<0.05). Flow cytometry showed that there were significant differences in the apoptosis, ROS and multicaspase level among ARPE-19 cell groups treated with different concentrations of ATRA ( F=86.39, 116.84, 101.40; all at P<0.01). The apoptosis rates of APRE-19 cells in the 2.5 μmol/L and 5 μmol/L of ATRA groups were significantly decreased than that of the blank control group, and the apoptosis rate of APRE-19 cells in the 10, 15 and 20 μmol/L of ATRA groups were significantly increaseded than that of the blank control group (all at P<0.01). The relative expression levels of multicaspase and ROS were significantly higher in the 2.5, 5, 10 and 20 μmol/L of ATRA groups than that of the blank control group (all at P<0.01). Western blot assay showed that the relative expression level of caspase 9 was increased in the 2.5, 5, 10, 15 and 20 μmol/L of ATRA groups than that of the blank control group (all at P<0.05). Compared with the blank control group, the relative expression levels of caspase 12 were increased in the 2.5 μmol/L of ATRA group and reduced gradually in the 5, 10, 15 and 20 μmol/L of ATRA groups, among which there were significant differences between the blank control group and 2.5, 15, and 20 μmol/L of ATRA groups (all at P<0.05). The relative expression level of caspase 3 was significantly increased in the 5, 10 and 20 μmol/L of ATRA groups than that of the blank control group (all at P<0.05). The relative expression level of cleaved caspase 3 was significantly increased in the 2.5, 5, 10, 15 and 20 μmol/L of ATRA groups than that of the blank control group (all at P<0.01). Quantitative real-time PCR assay showed that the relative expression levels of caspase 9 and caspase 12 mRNA were significantly higher in the 2.5, 5, 10 and 20 μmol/L of ATRA groups than that of the blank control group (all at P<0.01). The relative expression levels of caspase 3 mRNA were significantly higher in the 5 μmol/L and 10 μmol/L of ATRA groups than that of the blank control group (both at P<0.01). When the concentration of ATRA was lower than 10 μmol/L, the relative expression levels of caspase 9 and caspase 12 mRNA were elevated in a concentration-dependent manner.When the concentration of ATRA reached 20 μmol/L, the relative expression levels of caspase 9 and caspase 12 mRNA were markedly decreased, but it was still higher than that of the blank control group. Conclusions:ATRA induces apoptosis in ARPE-19 cells in vitro through activating the reactive oxygen species and endogenous caspase-dependent apoptotic pathway.
ABSTRACT
@#AIM: To investigate the endoplasmic reticulum stress(ERS)induced by all-trans retinoic acid(ATRA)in ARPE-19 cells.<p>METHODS:Immunofluorescence, real-time quantitative polymerase chain reaction and Western blot were used to detect the protein and mRNA expression of related signal pathways during the process of endoplasmic reticulum stress response induced by ATRA in ARPE-19 cells.<p>RESULTS: With the accumulation of ATRA concentration, the protein and mRNA levels of endoplasmic reticulum stress response marker proteins chop and BiP were significantly increased(<i>P</i><0.001); in the downstream signaling pathways, perk, eIF2 α, ATF4, IRE1 α and XBP1 were up-regulated(<i>P</i><0.001), while the expression of ATF6 did not change(<i>P</i>>0.05).<p>CONCLUSION: Over accumulation of ATRA induces ERS in ARPE-19 cells and activates PERK-EIF2 α-ATF4 and IRE1 α-XBP1 signaling pathways
ABSTRACT
Objective@#To research the functions of miR-3960 in all-trans-retinoic acid (atRA) induced cleft palate mouse model in order to provide the theoretical basis for gene therapy of cleft palate.@*Methods@#Excessive atRA induced cleft palate mouse model was established and palatine process tissues were collected. qRT-PCR was used to detect the expression of miR-3960. miRBase was used to analyse the characteristics of miR-3960 sequence. TargetScanMouse Prediction of microRNA targetswas used to predict the target genes of miR-3960. DAVID v6.8 database was used to perform bioinformatics analysis of target genes.@*Results@#miR-3960 was up-regulated in the experimental group. From the analysis of miRBase, we only got the sequences of miR-3960 in two species, and the two sequences were the same. There were 320 predicted target genes, the functions were mainly concentrated in cell proliferation, cell differentiation, embryo development and tissue development and so on (P<0.05), the signaling pathways were mainly concentrated in the calcium signaling pathway, cAMP signaling pathway and cGMP-PKG signaling pathway and so on (P<0.05).@*Conclusions@#In excessive atRA induced cleft palate mouse models, the up-regulated miR-3960 may result in cleft palate by inhibiting the proliferation and differentiation of palatal mesenchymal cells.
ABSTRACT
Objective: To investigate the effect of all-trans retinoic acid (ATRA) and vascular endothelial growth factor (VEGF) on the osteogenic differentiation of mouse embryonic fibroblasts (MEFs). Methods: The fetal mice in the uterus of NIH pregnant mice (pregnancy 12-15 days) were collected, and the heads and hearts etc. were removed. Then MEFs were separated from the rest tissues of the fetal mice and cultured by trypsin digestion and adherent culture. HEK-293 cells were used to obtain recombinant adenovirus-red fluorescent protein (Ad-RFP) and Ad-VEGF by repeatedly freezing and thawing. Alkaline phosphatase (ALP) staining and quantitative detection were used to detect the changes of ALP activity in MEFs applied with ATRA or VEGF alone or combined use of ATRA and VEGF on the 3rd and 5th days. The cultured 3rd to 4th generation MEFs were divided into groups A, B, C, and D, and were cultured with DMSO plus Ad-RFP, ATRA, Ad-VEGF, ATRA plus Ad-VEGF, respectively. Real-time fluorescence quantitative PCR (qRT-PCR) was used to detect the mRNA expressions of osteogenic markers including ALP, collagen type Ⅰ, osteopontin (OPN), osteocalcin (OCN), and angiogenic markers including VEGF, angiopoietin 1 (ANGPT1), and endomucin (EMCN) on the 3rd and 7th days. Immunohistochemical staining was used to detect the protein expressions of OPN and VEGF on the 3rd, 5th, and 7th days in each group. Alizarin red staining was used to detect calcium salt deposition levels in each group at 14 and 21 days after osteogenic induction. Fifteen athymic female nude mice aged 4 to 6 weeks were randomly divided into 3 groups and 5 mice in each group. Then MEFs treated with ATRA, Ad-VEGF, and ATRA plus Ad-VEGF were injected subcutaneously into the dorsal and ventral sides, respectively. X-ray observation, gross observation, and histological staining (Masson, HE, and Safranin O-fast green stainings) were performed at 5 weeks after implantation to observe the ectopic bone formation in nude mice in each group. Results: MEFs were successfully isolated and cultured. The acquired Ad-RFP and Ad-VEGF were successfully transfected into MEFs with approximately 50% and 20% transfection rates. ALP activity tests showed that ATRA or Ad-VEGF could enhance ALP activity in MEFs ( P<0.05), and ATRA had a stronger effect than Ad-VEGF; and the combined use of ATRA and Ad-VEGF significantly enhanced the ALP activity in MEFs ( P<0.05). qRT-PCR test showed that the combined use of ATRA and Ad-VEGF also increased the relative mRNA expressions of early-stage osteogenesis-related markers ALP, OPN, and collagen type I ( P<0.05); the relative mRNA expressions of angiogenesis-related markers VEGF, EMCN, and ANGPT1 increased at 7 days ( P<0.05). Immunohistochemical staining showed that ATRA combined with Ad-VEGF not only enhanced OPN protein expression, but also increased VEGF protein expression on 7th day. Alizarin red staining showed that the application of ATRA or Ad-VEGF induced weak calcium salt deposition, and the combined use of ATRA and Ad-VEGF significantly enhanced the effect of calcium salt deposition in MEFs. The results of implantation experiments in nude mice showed that X-ray films observation revealed obvious bone mass in the ATRA plus Ad-VEGF group, and the bone was larger than that in other groups. Histological staining showed a large amount of collagen and mature bone trabeculae, bone matrix formation, and gray-green collagen bone tissue, indicating that the combined use of ATRA and Ad-VEGF significantly enhanced the osteogenic effect of MEFs in vivo. Conclusion: The combined use of ATRA and VEGF can induce the osteogenic differentiation of MEFs.
ABSTRACT
[Abstract] Objective To investigate the effects of exclusive or joint application of all-trans retinoic acid (ATRA) and bone morphogenetic proteins (BMPs) on the osteocalcin (OCN) in mouse osteogenic precursor cells (MC3T3-E1) and bone marrow mesenchymal stem cells (BMSCs), and on the expressions of osteogenesis-related genes ALP, OCN and Runx2. Methods MC3T3-E1 cells and BMSCs were subcultured and stimulated with different concentrations of cytokines, and then divided into ten groups: control group, ATRA group, 5ng/ml BMP2/7 group, 50ng/ml BMP2/7 group, 5ng/ml BMP2 group, 50ng/ml BMP2 group, ATRA+5ng/ml BMP2/7 group, ATRA+50ng/ml BMP2/7 group, ATRA+5ng/ml BMP2 group, and ATRA+50ng/ml BMP2 group. The morphology of the cells was observed by immunofluorescence staining. The OCN expressions in the two kinds of cells were detected by ELISA. The expressions of ALP, OCN and Runx2 genes were detected by RT-qPCR. Results Immunofluorescence staining showed that the cells grew well and were structurally intact. Exclusive use of ATRA obviously inhibited the expressions of OCN in both MC3T3-E1 and BMSCs. On the 7th day, for MC3T3-E1 cells: ATRA alone group [(20.97±0.31)ng/ml] significantly decreased than control group [(33.45±0.55)ng/ml]; for BMSCs: ATRA alone group [(9.90±0.16)ng/ml] significantly decreased than control group [(14.30±0.53)ng/ml] (P<0.05). Exclusive application of BMPs promoted OCN expression in a concentration-dependent manner, and BMP2/7 showed a more strong promoting role. The OCN content in MC3T3-E1 cells treated with 50ng/ml BMP2/7 was (47.13±0.59)ng/ml, and in BMSCs was (49.92±0.53)ng/ml. 50ng/ml BMP2/7 completely antagonized the inhibitory effect of ATRA on cell OCN expression (P<0.05). RT-qPCR test showed that, for MC3T3-E1: ATRA alone significantly inhibited the expressions of ALP and OCN gene (P<0.05), but promoted the expression of Runx2 gene (P<0.05). BMPs alone promoted the expression of osteogenesis-related genes in a concentration-dependent manner (P<0.05). ATRA significantly inhibited the promoting role of BMPs on the OCN gene expression, but ATRA and 50ng/ml BMP2/7 synergistically promoted the expression of ALP gene (P<0.05); for BMSCs: ATRA alone significantly promoted the expression of Runx2 gene on the 7th day, but inhibited the expressions of ALP and OCN gene (P<0.05). 50ng/ml BMPs alone significantly promoted the expressions of ALP and Runx2 genes (P<0.05). 50ng/ml BMP2/7 antagonized the inhibitory effect of ATRA on ALP gene expression and promoted its expression (P<0.05). Conclusions ATRA alone may inhibit the expression of OCN in cells, and inhibits the expressions of ALP and OCN genes in cells; and BMP2/7 may significantly promote the expression of OCN in cells, and may promote the expression of osteogenesis-related genes; and 50ng/ml BMP2/7 may completely antagonize the osteogenic inhibition of ATRA on cells.
ABSTRACT
The present study was designed to analyze the metabolites of all-frans-retinal (atRal) and compare the cytotoxicity of atRal versus its derivative all-frans-retinoic acid (atRA) in human retinal pigment epithelial (RPE) cells. We confirmed that atRA was produced in normal pig neural retina and RPE. The amount of all-trans-retinol (atROL) converted from atRal was about 2.7 times that of atRal-derived atRA after incubating RPE cells with 10 µmol/L atRal for 24 h, whereas atRA in medium supernatant is more plentiful (91 vs. 29 pmol/mL), suggesting that atRA conversion facilitates elimination of excess atRal in the retina. Moreover, we found that mRNA expression of retinoic acid-specific hydroxylase CYP26b1 was dose-dependently up-regulated by atRal exposure in RPE cells, indicating that atRA inactivation may be also initiated in atRal-accumulated RPE cells. Our data show that atRA-caused viability inhibition was evidently reduced compared with the equal concentration of its precursor atRal. Excess accumulation of atRal provoked intracellular reactive oxygen species (ROS) overproduction, heme oxygenase-1 (HO-1) expression, and increased cleaved poly(ADP-ribose) polymerase 1 (PARP1) expression in RPE cells. In contrast, comparable dosage of atRA-induced oxidative stress was much weaker, and it could not activate apoptosis in RPE cells. These results suggest that atRA generation is an antidotal metabolism pathway for atRal in the retina. Moreover, we found that in the eyes of ABCA4−/−RDH8−/− mice, a mouse model with atRal accumulation in the retina, the atRA content was almost the same as that in the wild type. It is possible that atRal accumulation simultaneously and equally promotes atRA synthesis and clearance in eyes of ABCA4−/−RDH8−/− mice, thus inhibiting the further increase of atRA in the retina. Our present study provides further insights into atRal clearance in the retina.
ABSTRACT
Objective Decitabine (DAC) combined with the half dose priming regimen (HDPR) is a common treatment of myelodysplastic syndromes (MDS) and acute myeloid leukemia (AML) in the elderly. This study was to compare the clinical effect and safety of DAC combined with all-trans retinoic acid (ATRA) versus DAC plus HDPR in the treatment of MDS with excess of blasts (MDS-EB) or AML in elderly patients. Methods We retrospectively analyzed 48 elderly patients (≥60 years) with myeloid neoplasms (AML, MDS-EB-1 or MDS-EB-2) ineligible for standard chemotherapy treated in our hospital from January 2014 to October 2018, 22 by DAC+ATRA (group A) and the other 26 by DAC+HDPR (group B). We compared the overall response rate (ORR), overall survival (OS) and adverse events between the two groups of patients. Results No statistically significant difference was observed between groups A and B in ORR (86.4% vs 76.9%, P = 0.643) or median OS (26.2 vs 24.9 mo, P = 0.920). The median time to response was significantly longer in group A (2 courses) than in B (1 course) (P = 0.006). Compared with group A, group B showed remarkably lower incidence rates of grade-3 to -4 cytopenia (54.5% vs 84.6%, P = 0.029) and infection (45.5% vs 76.9%, P = 0.037), longer duration of neutropenia (P < 0.05), and higher volumes red blood cell infusion and platelet infusion (P < 0.05). There was no statistically significant difference in the incidence rate of bleeding between the two groups (P = 0.643). Conclusion DAC+ATRA and DAC+HDPR have comparable clinical effects on myeloid neoplasms in elderly patients, but the former is safer and better tolerated while the latter can achieve a more rapid response.
ABSTRACT
Histone lysine specific demethylase 1 (LSD1) has been recognized as an important modulator in post-translational process in epigenetics. Dysregulation of LSD1 has been implicated in the development of various cancers. Herein, we report the discovery of the hit compound (IC = 3.93 μmol/L) and further medicinal chemistry efforts, leading to the generation of compound (IC = 49 nmol/L, and = 16 nmol/L), which inhibited LSD1 reversibly and competitively with H3K4me2, and was selective to LSD1 over MAO-A/B. Docking studies were performed to rationalize the potency of compound . Compound also showed strong antiproliferative activity against four leukemia cell lines (OCL-AML3, K562, THP-1 and U937) as well as the lymphoma cell line Raji with the IC values of 1.79, 1.30, 0.45, 1.22 and 1.40 μmol/L, respectively. In THP-1 cell line, significantly inhibited colony formation and caused remarkable morphological changes. Compound induced expression of CD86 and CD11b in THP-1 cells, confirming its cellular activity and ability of inducing differentiation. The findings further indicate that targeting LSD1 is a promising strategy for AML treatment, the triazole-fused pyrimidine derivatives are new scaffolds for the development of LSD1/KDM1A inhibitors.
ABSTRACT
Background: All-trans retinoic acid (ATRA), a vitamin A-derived active metabolite, exerts important functions in hair biology. Previous studies indicated that excess ATRA hampered hair follicle morphogenesis and cyclic regeneration in adulthood, but other studies stated that ATRA promoted hair growth. Dermal papilla (DP), a cluster of specialized fibroblasts, plays pivotal roles in controlling development and regeneration of hair follicle. Several lines of evidence indicated that DP might be the target cells of ATRA in the hair follicle. To confirm this hypothesis, the present study was performed to explore the biological effects of ATRA on goat dermal papilla cells (DPCs) and clarify the roles of ATRA in hair biology. Results: Our experimental results indicated that key signaling transducers of ATRA were dynamically expressed in distinct stages of goat cashmere growth cycle, and high-dose ATRA treatment (10-5 M) significantly impaired the viability of goat DPCs and lowered the ratio of proliferating cells. Otherwise, goat DPCs were stimulated to enter apoptosis and their cell cycle progression was severely blocked by ATRA. Moreover, the expression of fibroblast growth factor 7 (Fgf7), one of the potent hair growth stimulators secreted by DPCs, was transcriptionally repressed following ATRA treatment. Conclusion: DPCs are the targets of ATRA in the hair follicle, and ATRA negatively regulates hair growth by the targeted suppression of cell viability and growth factor expression of goat DPCs. Through these observations, we offer a new mechanistic insight into the roles of ATRA in hair biology.
Subject(s)
Animals , Tretinoin/pharmacology , Goats , Hair Follicle/drug effects , Regeneration , In Vitro Techniques , Immunohistochemistry , Receptors, Retinoic Acid , Hair Follicle/cytology , Hair Follicle/growth & development , Cell Proliferation/drug effects , Fibroblast Growth Factor 7/genetics , Real-Time Polymerase Chain ReactionABSTRACT
OBJECTIVE: Recapitulation of the spermatogenesis process in vitro is a tool for studying the biology of germ cells, and may lead to promising therapeutic strategies in the future. In this study, we attempted to transdifferentiate Wharton's jelly-derived mesenchymal stem cells (WJ-MSCs) into male germ cells using all-trans retinoic acid and Sertoli cell-conditioned medium. METHODS: Human WJ-MSCs were propagated by the explant culture method, and cells at the second passage were induced with differentiation medium containing all-trans retinoic acid for 2 weeks. Putative germ cells were cultured with Sertoli cell-conditioned medium at 36℃ for 3 more weeks. RESULTS: The gene expression profile was consistent with the stage-specific development of germ cells. The expression of Oct4 and Plzf (early germ cell markers) was diminished, while Stra8 (a premeiotic marker), Scp3 (a meiotic marker), and Acr and Prm1 (postmeiotic markers) were upregulated during the induction period. In morphological studies, approximately 5% of the cells were secondary spermatocytes that had completed two stages of acrosome formation (the Golgi phase and the cap phase). A few spermatid-like cells that had undergone the initial stage of tail formation were also noted. CONCLUSION: Human WJ-MSCs can be transdifferentiated into more advanced stages of germ cells by a simple two-step induction protocol using retinoic acid and Sertoli cell-conditioned medium.
Subject(s)
Humans , Male , Acrosome , Biology , Germ Cells , In Vitro Techniques , Mesenchymal Stem Cells , Methods , Spermatocytes , Spermatogenesis , Tail , Transcriptome , TretinoinABSTRACT
All-trans retinoic acid (ATRA) is a highly effective treatment for acute promyelocytic leukemia (APL), a cytogenetically distinct subtype of acute myeloid leukemia (AML). However, ATRA-based treatment is not effective in other subtypes of AML. In non-APL AML, ATRA signaling pathway is impaired or downmodulated, and consequently fails to respond to pharmacological doses of ATRA. Therefore, complementary treatment strategies are needed to improve ATRA responsiveness in non-APL AML. In this study, we investigated the combined effect of ATRA and bromodomain inhibitor JQ1, proven to have potent anti-cancer activity mainly through inhibition of c-Myc. We showed that the combination of ATRA with JQ1 synergistically inhibited proliferation of AML cells. The synergistic growth inhibition was resulted from differentiation or apoptosis depending on the kind of AML cells. Concomitantly, the combined treatment of ATRA and JQ1 caused greater depletion of c-Myc and hTERT expression than each agent alone in AML cells. Taken together, these findings support the rationale for the use of the combination of ATRA and JQ1 as a therapeutic strategy for the treatment of AML.
Subject(s)
Apoptosis , Leukemia , Leukemia, Myeloid, Acute , Leukemia, Promyelocytic, Acute , TretinoinABSTRACT
All-trans retinoic acid (ATRA) has been studied for the treatment of cancer, including leukemia and breast cancer. This work aims to develop nanoemulsions (NE) loaded with a hydrophobic ion pair (HIP) of all-trans retinoic acid (ATRA) and a lipophilic amine, stearylamine (SA), and coated with hyaluronic acid (HA) to enhance anticancer activity and reducing toxicity. Blank NE was prepared by spontaneous emulsification and optimized prior to HIP incorporation. NE-ATRA was electrostatically coated with different concentrations of HA. Incorporation of ATRA-SA led to monodisperse NE with small size (129 ± 2 nm; IP 0.18 ± 0.005) and positive zeta potential (35.7 ± 1.0 mV). After coating with 0.5 mg/mL HA solution, the mean diameter slightly increased to 158 ± 5 nm and zeta potential became negative (-19.7 ± 1.2 mV). As expected, high encapsulation efficiency (near 100%) was obtained, confirmed by polarized light microscopy and infrared analysis. Formulations remained stable over 60 days and release of ATRA from NE was delayed after the hydrophilic HA-coating. HA-coated NE-ATRA was more cytotoxic than free ATRA for MDA-MB-231 and MCF-7 breast cancer cell lines, especially in the CD44 overexpressing cells. Blank coated formulations showed no cytotoxicity. These findings suggest that this easily-made HA-coated NE-ATRA formulation is a promising alternative for parenteral administration, thus improving the breast cancer therapy with this drug.
Subject(s)
Tretinoin/analysis , Breast Neoplasms/drug therapy , Pharmaceutical Preparations/analysis , Chemistry, Pharmaceutical , Hyaluronic AcidABSTRACT
Objective To investigate the induction of B-cell specific phenotype in classical Hodgkin lymphoma (cHL)upon all-trans retinoic acid(ATRA)incubation. Methods To construct B-cell specific promoter(CD19, CD79a,CD79b)driven reporter plasmid with NEO cassette to realize stable transfection and selection of cHL reporter cells. To verify the intact integration by amplification of the promoter and luciferase sequences,and to functionally validate the B-cell specific promoter by ABF1 interference and luciferase assay. Repoter cells were incubated with various doses of ATRA and luciferase activity was detected at 24,48 and 72 hours. Reporter cells were treated alone or in combination with 5-Aza and ATRA followed by luciferase assay. Endogenous B-cell specific genes(CD19, CD20, CD79a and CD79b) transcription and expression levels were detected by real-time PCR and immunoblot, respectively. The expression level of CD30 antigen on Hodgkin lymphoma cell membrane upon ATRA was assessed by flow cytometry. Results ATRA treatment stimulated B-cell specific signature in cHL cells including CD19,CD79a and CD79b while down-regulated their CD30 expression. Conclusions ATRA induces B-cell phenotype deficient cHL cells to regain their B-cell transcriptional program while abolishes their Hodgkin-specific machinery.