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Identification and expression profiles of genes and protens in SMMC-7721 cells
Xie Shun Feng1, Sun Shu Ming, Lu Xiao Feng
1Departments of General Surgery, The First Affiliated Hospital of Shantou University Medical College, Shantou, People's Republic of China.
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.
Insights
Cis-diammineplatinum (CDDP) treatment demonstrated anti-cancer effects in liver cancer cells SMMC-7721 by inducing apoptosis, altering cell cycle, and reducing motility. CDDP also inhibited tumor growth in mice, modulating key apoptosis-related genes and proteins.
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- Hepatocellular carcinoma (HCC) remains a significant global health challenge.
- Understanding the molecular mechanisms of chemotherapy agents is crucial for effective treatment strategies.
Purpose of the Study:
- To evaluate the effects of Cis-diammineplatinum (CDDP) on liver cancer cell apoptosis, motility, and cell cycle.
- To investigate the inhibitory effect of CDDP on tumor growth and its molecular mechanisms in a liver cancer mouse model.
Main Methods:
- In vitro assays: apoptosis rate, cell cycle analysis (flow cytometry), wound healing, transwell migration and invasion assays.
- In vivo studies: CDDP administration in hepatocellular carcinoma mice, blood biochemical analysis (WBC, total protein, albumin, A/G, ALT, AST, ALP).
- Molecular analysis: Immunohistochemistry (Fas, Bax, NF-κB, P53, Bcl-2), Real-time PCR (Bax mRNA, Bcl-2 mRNA).
Main Results:
- CDDP treatment increased apoptosis and S-phase cell cycle arrest in SMMC-7721 cells in a dose-dependent manner.
- CDDP reduced cell motility, invasion, and wound closure capacity of liver cancer cells.
- In vivo, CDDP modulated liver function markers and significantly altered the expression of pro-apoptotic (Fas, Bax) and anti-apoptotic (Bcl-2) proteins and genes, as well as NF-κB and P53.
Conclusions:
- CDDP exhibits significant anti-cancer effects on liver cancer cells SMMC-7721 in vitro, impacting apoptosis, motility, and cell cycle.
- CDDP demonstrates tumor growth inhibition in vivo, mediated by the modulation of key apoptosis-related signaling pathways and genes.
- These findings support CDDP as a potential therapeutic agent for hepatocellular carcinoma, warranting further investigation into its molecular mechanisms.
