Antiproliferative effect of mifepristone (RU486) on human neuroblastoma cells (SK-N-SH): in vitro and in vivo studies

RU486 (mifepristone), a glucocorticoid and progesterone receptor antagonist, has been reported to exert antiproliferative effects on tumor cells. Experiments were performed to analyze the effects of RU486 on the proliferation of the human neuroblastoma, both in vitro and in vivo, using the human neuroblastoma SK-N-SH cell line. The exposure in vitro of SK-N-SH cells to RU486 revealed a dose-dependent inhibition of 3H-thymidine incorporation due to a rapid but persistent inhibition of MAPKinase activity and ERK phosphorylation. A significant decrease of SK-N-SH cell number was evident after 3, 6, and 9 days of treatment (up to 40% inhibition), without evident cell death. The inhibitory effect exerted by RU486 was not reversed by the treatment of the cells with dexamethasone or progesterone. Moreover, RU486 induced a shift in SK-N-SH cell phenotypes, with an almost complete disappearance of the neuronal-like and a prevalence of the epithelial-like cell subtypes. Finally, the treatment with RU486 of nude mice carrying a SK-N-SH cell xenograft induced a strong inhibition (up to 80%) of tumor growth. These results indicated a clear effect of RU486 on the growth of SK-N-SH neuroblastoma cells that does not seem to be mediated through the classical steroid receptors. RU486 acted mainly on the more aggressive component of the SK-N-SH cell line and its effect in vivo was achieved at a concentration already used to inhibit oocyte implantation.

Mifepristone as pre-induction cervical ripening agent: a review article

Induction of labour after the period of viability by any methods medical, surgical or combined, for the purpose of vaginal delivery. The success of induction, to a great extent, depend upon pre-induction cervical status i.e. cervical ripening. So, ripening of cervix prior to induction i.e. pre-induction cervical ripening is one of the important steps for successful induction of labour. There are different methods for cervical ripening like prostaglandins (PGE). However, use of prostaglandins (PGE) and oxytocin as labour inducing agent has its own adverse effects on maternal and perinatal outcome. So, constant efforts are made for the less use of uterotonins. The present review aims to study the efficacy of oral Mifepristone for improvement in Bishop’s score, requirement of additional uterotonics, induction delivery interval, mode of delivery and neonatal outcome. Electronic databases were searched by using keywords ‘Mifepristone, RU486, PGE2 gel, Cervical ripening, Bishop’s score and Induction of labour’ and eleven articles were found from 2009 to 2018 which fulfils our study criteria and thus they were taken for review. Based on all the studies, Mifepristone appears to be effective cervical ripening in comparison to other agents with significant improvement in Bishop’s score, higher vaginal delivery rate, shorter induction delivery interval and good neonatal outcome.

Effect of RU486 on pre-adipocytes differentiation and NF-κB activation / 实用医学杂志

Objective To investigate the roles of RU486 inhibiting 3T3-L1 pre-adipocytes differentiation and regulating NF-κB activation. Methods Cells were treated with RU486 with concentrations of 0.1 ~ 10μmol/L for 48 h , then the relative contents of triglyceride were analyzed by Oil-Red O staining assay on 9 th day during adipogenesis. The mRNA expressions of PPARγ2,C/EBPa, LPL and FAS were further measured by Real-time PCR. IκBα protein level was detected by Western bolt and nuclear translocation of NF-κB was observed by immunofluorescence assay. Results The relative contents of triglyceride decreased with the increasing of RU486 concentration. Compared with the control, the relative contents of triglyceride in RU486-treatment groups from 0.5 μmol/L were significantly decreased (P < 0.05 or P < 0.01). Compared with the control, PPARγ2, C/EBPa, LPL and FAS mRNA expression and IκBα protein level were significantly decreased (P < 0.01) and NF-κB nuclear translocated from cytoplasm to nucleus in Group 5 mmol/L RU486. Conclusions RU486 could down regulate IκBα protein level , activate NF-κB nuclear translocation , then down regulate PPARγ2 , C/EBPa , LPL and FAS mRNA expression and inhibit adipocytes differentiation.

Ru486: del aborto químico a la contracepción de emergencia / Ru486: from chemical abortion to emergency contraception

El uso del mifepriston (RU486), en combinación con análogos de las prostaglandinas, en la inducción del aborto químico requiere una específica reflexión en relación a los principales aspectos farmacológicos y toxicológicos. La actual dialéctica bioética y biopolítica, a menudo ideologizada, impone aún más un tratamiento riguroso basado en evidencias científicas, para aclarar sobre todo los mecanismos de acción y de los eventos adversos. Estos últimos a veces también subvalorados o minimizados. Considerada la iniquidad del aborto voluntario, el artículo se propone también el objetivo de aclarar cómo, a la luz de una reciente bibliografía, el recurso al RU486 representa un significativo riesgo para la salud de las mujeres. Una particular atención está reservada a la aclaración etiopatogenética de las hemorragias y de las sepsis, en las cuales se han evidenciado también distintos decesos. En el artículo están presentes, además, los más actuales desarrollos de la investigación con RU486 sea para el tratamiento experimental de patologías –ginecológicas y no– como para el uso de la molécula de la contracepción hormonal y la contracepción de emergencias.

The use of mifepristone (RU486), in combination with prostaglandin analogues, in chemical abortion requires a specific reflection on the main aspects in pharmacology and toxicology. The current debate in bioethics and bio-policies, often ideological, imposes a more rigorous treatment based on scientific evidence, especially clarification of the mechanisms of action and severe adverse events. The latter is sometimes underestimated or minimized. Considering the inequity of voluntary abortion, this article aims also to clarify how, in the light of the most recent literature; the use of RU486 represents a significant risk to women’s health. Particular attention is given to etiopathogenetic clarification of bleeding and sepsis, which have also involved several deaths. The article reports the latest developments in research with RU486, whether for experimental treatment of pathologies –gynaecological and others– and for the use of the hormonal contraception molecule and emergency contraception.

Inducible IL-12 gene expression mediated by naked DNA containing an RU486 regulatory system / 中华微生物学和免疫学杂志

Objective To evaluate the inducibility of target gene expression induced by plasmid DNA carrying a RU486 regulatory system. Methods Plasmid pRS-LacZ encoding LacZ gene and plasmid pRS22 encoding murine interleukin-12(IL-12)gene were used to bansfect cells in vitro or be rapidly injected into the vein of mice. Then LacZ expression was assayed in cultured cells and tissues by X-gal staining.The IL-12 level was tested in the supernatants of cultured cells and in the serum with ELISA. ResultsIn the presence of RU486.30％ of the liver derived hepatic HepG2 cells were dyed blue while less than 1％of the HepG2 cells were blue colored in the absence of RU486. Less than 0.5％ of other non-hepatic cells were in blue with or without RU486. After transfection, the IL-12 level in the supernatants of HepG2 cells increased with increasing concentrations of RU486 treatment. After intravenous injection of plasmid pRS-LacZ,blue colored cells were observed only in liver tissue while induced by RU486. After intravenous injection of plasmid pRS22,the IL-12 expression level in serum was related with the dosage of RU486 or/and plasmid DNA. Conclusion Plasmid DNA containing RU486 regulatory system could open or close IL-12 gene expression by the addition or deletion of RU486. And the IL-12 expression level in serum could be regulated by altering the dose of plasmid DNA or/and RU486.

RU486-inducible and liver-specific expression of IL-12 gene in mice / 中华微生物学和免疫学杂志

Objective To investigate the inducible ability of plasmid DNA carrying a RU486 regulatory system.Methotis Plasmid pRS22 containing RU486 regulatory system,liver specific promoter and transgene IL-12 was injected into mice by hydrodynamic injection.RU486 was injected intraperitoneally into mice at difierent time points after plasmid administration.The IL-12 1evel in serum was tested by an ELISA kit.The distribution and inducible expression of pRS22 in mice were assayed by measuring DNA,RNA and protein levels by PCR,RT-PCR and immunohistochemical staining.Resuits To determine the duration of the activity of plasmid pRS22,mice injected with 10μg of pRS22 were treated repeatedly with 250μg/kg of RU486 per 7 days after hydrodynamic injection of plasmid.IL-12 expression in serum abruptly increased to peak was detected at 10 h after induction and declined to baseline on day 6.Though peak values of IL-12 decreased gradually after each induction,IL-12 in serum could be induced until 15 weeks after plasmid administration.A total of 5μg of pRS22 was injected into mice to detect the effect of different induction manners on the IL-12 expression.The mice were treated with RU486 per day or per 2 days iil 6 days,respectively.The induction per 2 days resulted in a wavelike pattern of serum IL-12 expression with peak lev-els on the day of induction alternating with lower values on the following day.In contrast,sustained levels of IL-12 could be achieved by administering RU486 per day.Plasmid DNA and GLp65 mRNA were detected in liver of mice with or without RU486 until at least day 28 after plasmid administration.However,IL-12 p35 mRNA was detected only in the liver of mice with RU486 induction.IL-12 immunohistochemical staining in liver demonstrated that IL-1 2 expressed predominantly in the hepatocytes near the surface of liver or between the central vein and portal area after induction with RU486.In contrast,no IL-12 expression was observed in the hepatocytes after induction with sesame oil.Conclusion Tight temporal and spatial control of transgene IL-12 expression could be achieved by RU486 regulatory system driven by liver specific promoter.

Effect of ovarian on FSH release and subunit synthesis in rat anterior pituitary cells in culture / 대한산부인과학회잡지

OBJECTIVE: to determine whether ovarian steroids directly regulate FSH release and B subunit synthesis at the pituitary level METHODS: In vitro study. After applying ovarian steroids in cultured rat anterior pituitary cells, we evaluate the amount of relesed FSH and FSH B subunit using dot-blot hybridization. RESULTS: Estradiol alone or GnRH-induced did not modulate FSH release and B ubunit synthesis. Although progesterone alone did not alter FSH release, GnRH-induced FSH release was significantly augmented by the treatment of progesterone. Progesterone alone stimulated FSH B mRNA level. Significant increase in FSH B mRNA level was also observed by the combined treatment of progesterone and GnRH. The effects of progesterone on GnRH-induced FSH release was significantly reduced by the treatment of progesterone antagonist, RU 486. The treatment with progesterone antagonist, RU 486 also abolished progesterone-induced FSH B subunit mRNA biosynthesis. CONCLUSION: From these results, it is, therefore, concluded that estradiol does not seem to be a major regulator for FSH synthesis but progesterone may exert its action at the pituitary level for the synthesis and release of FSH.

The responses of RU486 to the effects of corticosterone sulfate on cardiovascular neurons in the rostral ventrolateral medulla of rats / 中国药理学通报

0.05), but completely (3 neurons) or partially (9 neurons) blocked the excitatory effect induced by CORT. CONCLUSION CORT had rapid excitatory effects on the Caldiovascular neuronsin the RVLM. RU 486 had no responses to the baseline activity of the cardiovascular neurons, and but completely or partially blocked the effect of CORT on the cardiovascular neurons.

RU486 reverses dexamethasone inhibition of interleukin—2 prodnction and gene expression in lymphocytes of rat spleen / 中国免疫学杂志

By using ~3H—TdR incorporation and CTLL—2 cell line,we observed effects of RU486,anantagonist of type Ⅱ glucocorticoid receptor,on dexamethasone inhibition of lymphocyte prolifer-ation and interleukin—2(IL—2)production in rat spleen.It was shown that RU486 obviouslyantagonized the inhibitory actions of dexamethasone on lymphocyte proliferation and IL—2 pro-duction.By means of dot hybridization and Northern blot analysis,effects of dexamethasone andRU486 on IL—2 gene expression were investigated in lymphocytes of rat spleen.The datademonstrated that dexamethasone markedly decreased IL—2 mRNA production,RU486 alonedidn't affect IL-2 mRNA levels,but obviously reversed dexamethasone-mediated downregala-tion of IL—2 mRNA production in ConA—activated lymphocytes.These results suggest thatthe above effects of dexamethasone may be mediated by glucocorticoid receptor in lymphocytesof rat spleen.

INFLUENCE OF PROGESTERONE ANTAGONIST-RU486 ON THE MACROPHAGES IN THE UTERI OF PREGNANT MICE AND ITS RELEVANCE TO PREGNANCY / 解剖学报

Objective The experiment was conducted to explore the influence of progesterone antagonist\|RU486 on the macrophages in the uteri of pregnant mice and its relevance to pregnancy. Methods On day 4 and day10 of gestation, each mouse was subcutaneously injected with 150mg/L of RU486 (treatment group) or the same volume of saline (control group). The macrophages were immunohistochemically identified in the uterus 12, 24, 36?h after injection. Results Injection of RU486 on day 4 gestation had completely blocked the implantation of blastocysts. A large number of macrophages were found in the endometrium, myometrium and blood vessel layer 12～36?h after RU486 injection, which was significantly higher than that of the control groups ( P