ABSTRACT
Objective:To investigate the effects of cell adhesion molecule 1 (CADM1) promoter methylation in ovarian cancer on gene transcription and protein expression levels, and the regulation mechanism of mirNA-148A on CADM1 methylation levels.Methods:A total of 86 patients with ovarian cancer who received surgical treatment in the Affiliated Hospital of Hangzhou Normal University from Jun. 2018 to Jun. 2020 were selected as study subjects. The methylation level of CADM1 gene CpG island in ovarian cancer tissues and adjacent tissues was quantitatively detected. Quantitative real-time polymerase chain reaction was used to detect the mRNA and mirNA-148a expressions of CADM1 gene. The CADM1 gene and DNMT1 protein levels were detected by Western blot. Human ovarian cancer SKOV3 cells were treated with different concentrations of methyltransferase inhibitors (5-Azacytidine, 5-aza) , and CADM1 mRNA expression was detected 72 h later. Human ovarian cancer cell lines SKOV3 were transfected with mir-335-5p mimic, inhibitor and negative control respectively. Then mir-335-5p expression level and CADM1 gene methylation level were detected after transfection.Results:The methylation level of CADM1-1 island in ovarian cancer tissues was 2.89%±0.82%, significantly higher than that of paracancerous normal tissues 1.86%±0.68% ( t=4.936, P<0.001) , and that of CADM1-2 island in ovarian cancer tissues was 3.12%±0.93%, significantly higher than that of paracancerous normal tissues (2.27%±0.69%, t=5.114, P<0.001) . Pearson correlation analysis showed that the methylation level of CADM1-1 island and CADM1-2 island in ovarian cancer tissues was significantly negatively correlated with the relative mRNA expression (r was -0.615 and -0.582, respectively, and both P<0.001) , and with the protein expression level of CADM1 (r was -0.521 and -0.612, respectively, and both P<0.001) . The relative expression level of mirNA-148a in ovarian cancer tissues was 1.53±0.42, significantly lower than that in paracancer tissues (2.59±0.73, t=6.113, P<0.001) . After treatment with different concentrations of 5-AZA, mRNA expression levels of CADM1 gene in SKOV3 cells were significantly higher in the low concentration group and the high concentration group than in the control group (both P<0.05) , and mRNA expression levels in the high concentration group were significantly higher than in the low concentration group ( P<0.05) . After mirNA-148A transfected SKOV3 cells, the relative expression levels of mirNA-148a in the mimic group were significantly increased, while those of inhibitor group were significantly decreased ( P<0.001) . The DNMT1 expression level and CADM1 gene methylation level of mimic group were significantly decreased, while those of inhibitor group were significantly increased (P<0.001) . Conclusion:In ovarian cancer, miRNA-148a can regulate the DNA methylation level of CADM1 gene by acting on the downstream target protein DNMT1, thus affecting the mRNA and protein expression levels of CDM1 gene and participating in the pathogenesis of ovarian cancer.
ABSTRACT
【Objective】To observe the effect of plasma ?-endogenous peptides (?-EP) on gestational function in endometriosis(EMT) rats.【Methods】 Female SD rats at the third estrous cycle were used to establish EMT model.Then 20 EMT rats were equally randomized into the model group and naltrexone(an opioid receptor antagonist) group,and other 10 rats were in the pseudo-operation group.Naltrexone group was given naltrexone 2?mg?kg~-1?d~-1 by gastric gavage during proestrus and normal saline at other time,and the model group and pseudo-operation group were given normal saline for one month.Serum levels of ?-EP,luteinizing hormone(LH) and prolactin(PRL) were examined during proestrus,and serum progesterone(P) level and gestational function were observed after estrus.【Results】Serum levels of ?-EP and PRL increased and LH decreased during proestrus,and serum P level also decreased after estrus in the model group(P
ABSTRACT
【Objective】To investigate the therapeutic effect of Qiangxin Prescription(QP)on rabbits with heart failure induced by volume overload.【Methods】Twenty New Zealand male rabbits were randomized into four groups:the control group,QP group(QP 2.6?g/kg),western medicine group(Digoxin 20??g/kg and furosemide 1mg/kg)and QP+western medicine group(QP 2.6?g/kg,Digoxin 20??g/kg and furosemide 1?mg/kg).Rabbit models with heart failure induced by volume overload were established.Six hours after the modeling,the treatment groups were given the corresponding drugs according to the experimental design,the treatment lasting 7 days.Four-channel physiologic instrument was used to examine the heart rate(HR),aortic systolic blood pressure(SBP),aortic diastolic blood pressure(DBP),left ventricular systolic pressure(LVSP),left ventricular diastolic pressure(LVDP),the maximum rate of left ventricular pressure rise(+dp/dt_(max)),the maximum rate of left ventricular pressure decrease(-dp/dt_(max))and the maximum rate of left ventricular pressure change(?dp/dt_(max)).Meanwhile,the serum contents of superoxide dismustase(SOD)and malondialdehyde(MDA)were detected.【Results】The difference was insignificant in the body weight in different groups after the modeling but the body weight in western medicine group was decreased one week after medication(P