ABSTRACT
Objective To establish the expression profiles of microRNA (miRNA) in SMMC-7721 and CCC-HEL-1 cell lines in order to provide new clues to study the mechanism of miRNA function in hepatocellular carcinoma (HCC) and its treatment. Methods SMMC-7721 and CCC-HEL-1 cell lines were cultured in vitro. Total RNAs were extracted by TRIzol, followed by RNA quantification and quality control. The RNAs were used to detect the expression of miRNA in these two cell lines by miRNA array. The miRNA of interest was then verified by real-time PCR. Results In total,238 differentially expressed miRNAs were found, including 154 overexpressed and 84 underexpressed miRNAs. 64 miRNAs were upregulated more than 4 times and26 miRNAs were upregulated more than 10 times. 22 miRNAs were downregulated more than 4 times 10 miRNAs weredownregulated more than 5times, and 1 miRNA was downregulated more than 10 times. Real-time PCR validation suggested that has-miR-205 and has-let-7f in liver cancer increased by 2. 7 and 2. 3 times, respectively.Conclusion There are differences in the expression of miRNA between the SMMC-7721 and CCC-HEL-1 cell lines and the profiles of miRNA expression were established for both cell lines. It was found that has-miR-205 and has-let-7f were upregulated in liver cancer.
ABSTRACT
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.
ABSTRACT
Objective To explore the expression and significance of transforming growth factor-?(TGF-?),epidermal growth factor receptor (EGFR) and proliferation-related antigen Ki-67 in nephroblastoma. Methods Immunohistochemistry method was used to detect the expression of TGF-?,EGFR and Ki-67 in 8 cases of normal kidney,35 cases of nephroblastoma and 14 cases of the tissues adjacent to nephroblastoma.The differences of their expression were compared among the 3 kinds of tissues and different pathologic characteristics of nephroblastoma. Results The expression rates of TGF-?,EGFR and Ki-67 in nephroblastoma were 51.4%(18/35),62.9%(22/35) and 68.6%(24/35),respectively,which were significantly higher than those in the tissues adjacent to nephroblastoma and the normal kidney;the differences were significant (P0.05);but positive relationship was found between the expression and clinical stages (P