1.Effects of TSA on promoter methylation and expression of E-cadherin gene in hepatocellular carcinoma cell lines
Jiaxiang WANG ; Guangcheng GUO ; Qiuliang LIU ; Zhengjun FAN ; Bin DAI
Chinese Journal of General Surgery 2009;24(3):230-233
Objective To study the effects of histone deacertylase inhibitor (TSA) on promoter methylation and expression of E-cadherin gene in a hepatocellular carcinoma cell line SMMC7721. Methods Hepatocellular carcinoma cell line SMMC-7721 was treated with TSA (300 nm/L), MTT method was used to investigate the growth inhibition ratio, TUNNL was conducted to measure the apoptosis ratio, methylation-specific PCR (MSP) was employed to detect changes in the CpG island methylation of E-cad promoter region, Western blot technique was used to detect the expression of E-cad gene and DNMT3b before and after TSA treatment, respectively. Results TSA decreases the SMMC-7721 cell viability and induces apoptosis, the growth inhibition ratio was 21.85% compared with control group. The apoptosis ratio of control group was (4.69±0.56)% ,the apoptosis ratio of TSA treatment group was (14.94±0.91)%. The apoptosis ratio of TSA treatment group was significantly higher than that of control group(P = 0.000). Before treated with TSA, the CpG island of E-cad promoter region was methylated, and the expression of E-cad was negative. TSA treatment induces demethylation of the CpG island in E-cad promoter region, causes the re-expression of E-cad. TSA reduces the expression of DNMT3b. Conclusions TSA decreases the SMMC-7721 cell viability and induces apoptosis, reverses the methylation status of E-cad promoter region, and resumes E-cad gene expression. TSA may induce demethylation through down-regulating the expression of DNMT3b.
2.The inhibitory effect of eupatorium japonicum thunb and foeniculum vulgare extract on prostatic hyperpla-sia in rats
Wei TANG ; Guangcheng DAI ; Boxin XUE ; Yuxi SHAN ; Wenfang ZHANG
Journal of Medical Postgraduates 2014;(12):1266-1268
Objective Benign prostatic hyperplasia ( BPH) is one of common diseases in aged males , and searching for new therapeutic drugs to BPH has been a research hotspot in recent years .This article was to study the inhibitory effect of eupatorium ja-ponicum thunb and foeniculum vulgare extract ( EFE) on benign prostatic hyperplasia in rats and its possible mechanism . Methods 48 male rats were randomly divided into 6 groups:normal control group without any treatment , model group of BPH treated with subcu-taneous injection of testosterone propionate , positive control group of BPH treated with dutasteride , high, middle and low dosage groups according to different EFE dosage (156 mg/kg, 234 mg/kg and 312 mg/kg).45 days after the treatment, the rats were sacrificed for measurement of the prostate glandular wet weight , the index of prostate gland ( PI ) , the morphological changes of prostate gland by light microscopy and the content of sex hormone . Results The prostate wet weight and PI decreased after EFE treatment for 45 days compared with the BPH model group(P<0.01 ).The hyperplastic glandular epithelium papilla waned and even disappeared in three EFE groups under the light microscope , and the epithelial cells became cubical or flat .High dosage EFE group (312 mg/kg) has simi-lar efficacy to dutasteride group .EFE significantly reduced serum testosterone content , dihydrotestosterone content and T/E2 ratio( P<0.05 ). Conclusion EFE can significantly inhibit prostatic hyperplasia in rats , and its mechanism is related to the decrease of the contents of serum testosterone and dihydrotestosterone as well as T/E2 ratio.
3.Circulating miRNA-141 as a non-invasive biomarker for prostate cancer detection and prognosis
Yufeng LIAO ; Jinhua DAI ; Qifeng MAO ; Zhankun ZHU ; Guangcheng JIA
Chinese Journal of Pathophysiology 2014;(10):1887-1890
AIM:To analyze circulating miR-141 in the serum as a non-invasive biomarker in the patients with prostate cancer ( PCa) and benign prostate hyperplasia ( BPH) , and healthy individuals.METHODS: A total of 75 pa-tients with PCa, 52 with BPH and 40 healthy individuals were enrolled into this study.Total RNA was isolated from the se-rum samples and the circulating levels of miR-141 were determined using quantitative real-time polymerase chain reaction. RESULTS:The serum levels of miR-141 were significantly higher in the patients with PCa compared to the patients with BPH and the healthy controls (P<0.01).The level of miR-141 in PCa group obviously differed from that in BPH group and healthy control group with high diagnosis performance, with areas under the curve of 0.785 and 0.801, respectively. No statistically significant difference of the serum miR-141 levels between the patients with BPH and healthy individuals was observed (P>0.05).The serum miR-141 level was also found to be related to Gleason score, clinical stage and bone me-tastasis status of the patients with PCa (P<0.05), and the patients with higher Gleason scores had higher serum miR-141 levels.No relationship was detected between miRNA-141 level and the patient’ s age, biochemistry recurrence and serum prostate-specific antigen level (P>0.05 for all comparisons).CONCLUSION: Circulating miR-141 could serve as a non-invasive biomarker for prostate cancer diagnosis, staging and prognosis prediction.