Identification and expression profiles of genes and protens in SMMC-7721 cells

Mol Biol Rep. 2014 Feb;41(2):987-96. doi: 10.1007/s11033-013-2943-2. Epub 2014 Jan 9.

Abstract

In the study presented here, we first evaluated effect of CDDP on liver cancer cells SMMC-7721 apoptosis and motility capacity. Then, we evaluate inhibitory effect of CDDP on tumour growth and its possible molecular mechanism in liver cancer mice model. Results showed that the apoptosis rate of cells decreased with increasing CDDP. Analysis of the effect of the CDDP on cell cycle was performed by flow cytometry and results show a dose-dependent increase in the percentage of cells in the S-phase of the cell cycle, with a decrease in the percentage of cells in the G1 and G2/M phases. CDDP did not close the wound even after 48 h, as opposed to untreated cells (0 mg/l). Similarly, the migratory and invasion capacity of SMMC-7721 cells was also reduced after treatment with CDDP, as evaluated by a transwell assay. Animal experiment indicated that CDDP administration could increase blood WBC, total protein, albumin and A/G, decrease blood alanine aminotransferase, aspartate aminotransferase and alkaline phosphatase levels in hepatocellular carcinomas mice. Immunohistochemistry analysis showed that positive expression of Fas and Bax proteins in the medicine-treated (II, III) group was significantly higher, whereas the expression of NF-κB, P53, Bcl-2 proteins was significantly lower than those of the control group. Gene expression analysis using Real time PCR methods revealed a significant up-regulation in the expression levels of Bax mRNA in the medicne-treated (II, III) group when compared to untreated control. In contrast, CDDP-treated group showed a significant down regulation in the expression levels of Bcl-2 mRNA as compared to untreated control group. These results are in agreement with immunohistochemistry data. Our observations indicate that CDDP has damaged effects on liver tumour cells SMMC-7721 including apoptosis, motility and cell cycle under in vitro. CDDP can enhance pro-apoptosis gene Fas, Bax expression, decrease anti-apoptosis genes Bcl-2 expression, and mutant genes P53, NF-κB proteins expression.

MeSH terms

  • Animals
  • Apoptosis / drug effects
  • Carcinoma, Hepatocellular / genetics*
  • Cell Proliferation / drug effects
  • Cisplatin / administration & dosage*
  • Cisplatin / pharmacology
  • G2 Phase / drug effects
  • Gene Expression Regulation, Neoplastic / drug effects*
  • Liver Neoplasms / genetics*
  • Liver Neoplasms / pathology
  • Mice
  • NF-kappa B / metabolism
  • Proto-Oncogene Proteins c-bcl-2 / biosynthesis
  • bcl-2-Associated X Protein / biosynthesis

Substances

  • NF-kappa B
  • Proto-Oncogene Proteins c-bcl-2
  • bcl-2-Associated X Protein
  • Cisplatin