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Sorafenib suppresses the epithelial-mesenchymal transition of hepatocellular carcinoma cells after insufficient radiofrequency ablation.

Dong S, Kong J, Kong F, Kong J, Gao J, Ji L, Pan B, Chen L, Zheng L, Sun W - BMC Cancer (2015)

Bottom Line: An ectopic nude mice model was used to evaluate the effect of sorafenib on the growth of HepG2 cells in vivo after insufficient RFA.Higher expression of PCNA, Ki67, N-cadherin, MMP-2 and MMP-9, was also observed in HepG2-H tumors.Sorafenib blocked the enhanced growth of HepG2 cells in vivo after insufficient RFA.

View Article: PubMed Central - PubMed

Affiliation: Department of Hepatobiliary Surgery, Beijing Chao-yang Hospital, Capital Medical University, Beijing, 100043, China. shuying19870219@163.com.

ABSTRACT

Background: Epithelial-mesenchymal transition (EMT) played an important role in the progression of hepatocellular carcinoma (HCC) after insufficient radiofrequency ablation (RFA). However, whether sorafenib could be used to suppress the EMT of HCC after insufficient RFA and further prevent the progression of residual HCC remains poorly unknown.

Methods: Insufficient RFA was simulated using a water bath (47 °C 5, 10, 15, 20 and 25 min gradually). MTT assay and transwell assay were used to evaluate the effects of sorafenib on viability, migration and invasion of HepG2 and SMMC7721 cells after insufficient RFA in vitro. After insufficient RFA, the molecular changes in HCC cells with the treatment of sorafeinb were evaluated using western blot and ELISAs. An ectopic nude mice model was used to evaluate the effect of sorafenib on the growth of HepG2 cells in vivo after insufficient RFA.

Results: HepG2 and SMMC7721 cells after insufficient RFA (named as HepG2-H and SMMC7721-H) exhibited enhanced viability, migration and invasion in vitro. Sorafenib inhibited the enhanced viability, migration and invasion of HepG2 and SMMC7721 cells after insufficient RFA. Molecular changes of EMT were observed in HepG2-H and SMMC7721-H cells. Sorafenib inhibited the EMT of HepG2-H and SMMC7721-H cells. HepG2-H cells also exhibited larger tumor size in vivo. Higher expression of PCNA, Ki67, N-cadherin, MMP-2 and MMP-9, was also observed in HepG2-H tumors. Sorafenib blocked the enhanced growth of HepG2 cells in vivo after insufficient RFA.

Conclusions: Sorafenib inhibited the EMT of HCC cells after insufficient RFA, and may be used to prevent the progression of HCC after RFA.

No MeSH data available.


Related in: MedlinePlus

Sorafenib suppressed the EMT of HCC cells after insufficient RFA. Gray analysis of all bands was used to quantify expression levels of MMP-2, MMP-9, E-cadherin, N-cadherin, vimentin, snail, p-Akt, Akt, p-ERK1/2 and ERK1/2 in HepG2 and HepG2-H cells (a) The concentration of cytokines secreted into the conditioned medium of HepG2 and HepG2-H cells was detected by ELISA analysis (b). Error bars represent the SEM of data obtained in three independent experiments. P value <0.05 was considered statistically significant; **p < 0.01, ***p < 0.001
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Fig4: Sorafenib suppressed the EMT of HCC cells after insufficient RFA. Gray analysis of all bands was used to quantify expression levels of MMP-2, MMP-9, E-cadherin, N-cadherin, vimentin, snail, p-Akt, Akt, p-ERK1/2 and ERK1/2 in HepG2 and HepG2-H cells (a) The concentration of cytokines secreted into the conditioned medium of HepG2 and HepG2-H cells was detected by ELISA analysis (b). Error bars represent the SEM of data obtained in three independent experiments. P value <0.05 was considered statistically significant; **p < 0.01, ***p < 0.001

Mentions: HepG2-H and SMMC7721-H displayed a spindle shape with less cell-cell adhesion and increased formation of pseudopodia, and sorafenib inhibited the morphological changes above (Fig. 3a). To evaluate whether EMT had occurred in HepG2-H and SMMC7721-H cells, EMT markers were examined. Western blot and ELISA assay showed that significant reduction in E-cadherin expression and up-regulation of N-cadherin, vimentin, snail, MMP-2 and MMP-9 were found in HepG2-H (Figs. 3b and 4) and SMMC7721-H cells (Fig. 3b and Additional file 2: Figure S3). Sorafenib increased the expression of E-cadherin, and decreased the expression of N-cadherin, vimentin, snail, MMP-2 and MMP-9 in HCC cells in a dose-dependent manner (Figs. 3b, 4 and Additional file 2: Figure S3). To explore the signaling mechanisms involved in the sorafenib on EMT of HCC cells after insufficient RFA, we tested Akt and ERK1/2 signaling pathways. HepG2-H and SMMC7721-H cells showed significantly increased expression of p-Akt and p-ERK1/2 compared with HepG2 and SMMC7721 cells respectively (Figs. 3c, 4a and Additional file 2: Figure S3A). Furthermore, sorafenib inhibited the expression of p-Akt and p-ERK1/2 in HCC cells in a dose-dependent manner (Figs. 3c, 4a and Additional file 2: Figure S3A).Fig. 3


Sorafenib suppresses the epithelial-mesenchymal transition of hepatocellular carcinoma cells after insufficient radiofrequency ablation.

Dong S, Kong J, Kong F, Kong J, Gao J, Ji L, Pan B, Chen L, Zheng L, Sun W - BMC Cancer (2015)

Sorafenib suppressed the EMT of HCC cells after insufficient RFA. Gray analysis of all bands was used to quantify expression levels of MMP-2, MMP-9, E-cadherin, N-cadherin, vimentin, snail, p-Akt, Akt, p-ERK1/2 and ERK1/2 in HepG2 and HepG2-H cells (a) The concentration of cytokines secreted into the conditioned medium of HepG2 and HepG2-H cells was detected by ELISA analysis (b). Error bars represent the SEM of data obtained in three independent experiments. P value <0.05 was considered statistically significant; **p < 0.01, ***p < 0.001
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4663721&req=5

Fig4: Sorafenib suppressed the EMT of HCC cells after insufficient RFA. Gray analysis of all bands was used to quantify expression levels of MMP-2, MMP-9, E-cadherin, N-cadherin, vimentin, snail, p-Akt, Akt, p-ERK1/2 and ERK1/2 in HepG2 and HepG2-H cells (a) The concentration of cytokines secreted into the conditioned medium of HepG2 and HepG2-H cells was detected by ELISA analysis (b). Error bars represent the SEM of data obtained in three independent experiments. P value <0.05 was considered statistically significant; **p < 0.01, ***p < 0.001
Mentions: HepG2-H and SMMC7721-H displayed a spindle shape with less cell-cell adhesion and increased formation of pseudopodia, and sorafenib inhibited the morphological changes above (Fig. 3a). To evaluate whether EMT had occurred in HepG2-H and SMMC7721-H cells, EMT markers were examined. Western blot and ELISA assay showed that significant reduction in E-cadherin expression and up-regulation of N-cadherin, vimentin, snail, MMP-2 and MMP-9 were found in HepG2-H (Figs. 3b and 4) and SMMC7721-H cells (Fig. 3b and Additional file 2: Figure S3). Sorafenib increased the expression of E-cadherin, and decreased the expression of N-cadherin, vimentin, snail, MMP-2 and MMP-9 in HCC cells in a dose-dependent manner (Figs. 3b, 4 and Additional file 2: Figure S3). To explore the signaling mechanisms involved in the sorafenib on EMT of HCC cells after insufficient RFA, we tested Akt and ERK1/2 signaling pathways. HepG2-H and SMMC7721-H cells showed significantly increased expression of p-Akt and p-ERK1/2 compared with HepG2 and SMMC7721 cells respectively (Figs. 3c, 4a and Additional file 2: Figure S3A). Furthermore, sorafenib inhibited the expression of p-Akt and p-ERK1/2 in HCC cells in a dose-dependent manner (Figs. 3c, 4a and Additional file 2: Figure S3A).Fig. 3

Bottom Line: An ectopic nude mice model was used to evaluate the effect of sorafenib on the growth of HepG2 cells in vivo after insufficient RFA.Higher expression of PCNA, Ki67, N-cadherin, MMP-2 and MMP-9, was also observed in HepG2-H tumors.Sorafenib blocked the enhanced growth of HepG2 cells in vivo after insufficient RFA.

View Article: PubMed Central - PubMed

Affiliation: Department of Hepatobiliary Surgery, Beijing Chao-yang Hospital, Capital Medical University, Beijing, 100043, China. shuying19870219@163.com.

ABSTRACT

Background: Epithelial-mesenchymal transition (EMT) played an important role in the progression of hepatocellular carcinoma (HCC) after insufficient radiofrequency ablation (RFA). However, whether sorafenib could be used to suppress the EMT of HCC after insufficient RFA and further prevent the progression of residual HCC remains poorly unknown.

Methods: Insufficient RFA was simulated using a water bath (47 °C 5, 10, 15, 20 and 25 min gradually). MTT assay and transwell assay were used to evaluate the effects of sorafenib on viability, migration and invasion of HepG2 and SMMC7721 cells after insufficient RFA in vitro. After insufficient RFA, the molecular changes in HCC cells with the treatment of sorafeinb were evaluated using western blot and ELISAs. An ectopic nude mice model was used to evaluate the effect of sorafenib on the growth of HepG2 cells in vivo after insufficient RFA.

Results: HepG2 and SMMC7721 cells after insufficient RFA (named as HepG2-H and SMMC7721-H) exhibited enhanced viability, migration and invasion in vitro. Sorafenib inhibited the enhanced viability, migration and invasion of HepG2 and SMMC7721 cells after insufficient RFA. Molecular changes of EMT were observed in HepG2-H and SMMC7721-H cells. Sorafenib inhibited the EMT of HepG2-H and SMMC7721-H cells. HepG2-H cells also exhibited larger tumor size in vivo. Higher expression of PCNA, Ki67, N-cadherin, MMP-2 and MMP-9, was also observed in HepG2-H tumors. Sorafenib blocked the enhanced growth of HepG2 cells in vivo after insufficient RFA.

Conclusions: Sorafenib inhibited the EMT of HCC cells after insufficient RFA, and may be used to prevent the progression of HCC after RFA.

No MeSH data available.


Related in: MedlinePlus