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The aPKCι blocking agent ATM negatively regulates EMT and invasion of hepatocellular carcinoma.

Ma CQ, Yang Y, Wang JM, Du GS, Shen Q, Liu Y, Zhang J, Hu JL, Zhu P, Qi WP, Qian YW, Fu Y - Cell Death Dis (2014)

Bottom Line: Our study showed that EMT took place in MMH-R cells under the effect of transforming growth factor-β1 (TGF-β1) overexpressing aPKCι.In conclusion, our result suggested that aPKCι could be an important bio-marker of tumor EMT, and used as an indicator of invasion and malignancy.ATM might be a promising agent for targeted treatment of HCC.

View Article: PubMed Central - PubMed

Affiliation: Department of Biliary and Pancreatic Surgery/Cancer Research Center Affiliated Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.

ABSTRACT
Epithelial-to-mesenchymal transition (EMT) has an important role in invasion and metastasis of hepatocellular carcinoma (HCC). To explore the regulatory mechanism of atypical protein kinase C ι (aPKCι) signaling pathways to HCC development, and find an agent for targeted therapy for HCC, immortalized murine hepatocytes were employed to establish an EMT cell model of HCC, MMH-RT cells. Our study showed that EMT took place in MMH-R cells under the effect of transforming growth factor-β1 (TGF-β1) overexpressing aPKCι. Furthermore, we showed that the aPKCι blocking agent aurothiomalate (ATM) inhibited EMT and decreased invasion of hepatocytes. Moreover, ATM selectively inhibited proliferation of mesenchymal cells and HepG2 cells and induced apoptosis. However, ATM increased proliferation of epithelial cells and had little effect on apoptosis and invasion of epithelial cells. In conclusion, our result suggested that aPKCι could be an important bio-marker of tumor EMT, and used as an indicator of invasion and malignancy. ATM might be a promising agent for targeted treatment of HCC.

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Related in: MedlinePlus

Apoptosis of MMH-R and MMH-RT induced by ATM was determined by flow cytometry. At 48-h ATM incubation in a range of 1–600 μmol/l, the apoptotic rates of MMH-R and MMH-RT cells were detected by flow cytometry. (a) Typical FACS schematic. (b) Following the increased ATM concentration, the standardized apoptotic rates (%) of mesenchymal MMH-RT cells were dramatically higher than those of epithelial MMH-R cells when the concentration of ATM treatment was greater than 200 μmol/l (n=3, Student's t-test, mean±S.D., *P<0.05). (c) Expression of caspase 3 (cleaved-caspase 3, pro-caspase 3) in MMH-R and MMH-RT cells was detected by western blotting (n=3, Student's t-test, mean±S.D., *P<0.05, **P<0.01, ImageJ2X software)
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fig5: Apoptosis of MMH-R and MMH-RT induced by ATM was determined by flow cytometry. At 48-h ATM incubation in a range of 1–600 μmol/l, the apoptotic rates of MMH-R and MMH-RT cells were detected by flow cytometry. (a) Typical FACS schematic. (b) Following the increased ATM concentration, the standardized apoptotic rates (%) of mesenchymal MMH-RT cells were dramatically higher than those of epithelial MMH-R cells when the concentration of ATM treatment was greater than 200 μmol/l (n=3, Student's t-test, mean±S.D., *P<0.05). (c) Expression of caspase 3 (cleaved-caspase 3, pro-caspase 3) in MMH-R and MMH-RT cells was detected by western blotting (n=3, Student's t-test, mean±S.D., *P<0.05, **P<0.01, ImageJ2X software)

Mentions: ATM-induced apoptosis in 48 h was detected by fluorescence activated cell sorter (FACS) (Figure 5a). The results showed that the standardized apoptotic rate (%) of mesenchymal MMH-RT cells was dramatically higher than that of epithelial MMH-R cells when concentration of ATM treatment was above 200 μmol/l (Figure 5b, Student's t-test, P<0.05, Supplementary Figure 2), while in MMH-R cells, the apoptosis rate had no obvious correlation with concentration (Figure 5b; Supplementary Figure 2). We also detected the expression of caspase 3, a key molecule of apoptosis signal pathway, in the MMH-R and MMH-RT cells. The results exhibited that the expression of cleaved-caspase 3 of MMH-RT cells was significantly higher than that of MMH-R cells with the increased concentration of ATM (Figure 5c, Student's t-test, P<0.01). These findings indicated that the aPKCι signaling inhibitor could activate the caspase 3-dependent apoptosis pathway and significantly promote the apoptosis of MMH-RT cells.


The aPKCι blocking agent ATM negatively regulates EMT and invasion of hepatocellular carcinoma.

Ma CQ, Yang Y, Wang JM, Du GS, Shen Q, Liu Y, Zhang J, Hu JL, Zhu P, Qi WP, Qian YW, Fu Y - Cell Death Dis (2014)

Apoptosis of MMH-R and MMH-RT induced by ATM was determined by flow cytometry. At 48-h ATM incubation in a range of 1–600 μmol/l, the apoptotic rates of MMH-R and MMH-RT cells were detected by flow cytometry. (a) Typical FACS schematic. (b) Following the increased ATM concentration, the standardized apoptotic rates (%) of mesenchymal MMH-RT cells were dramatically higher than those of epithelial MMH-R cells when the concentration of ATM treatment was greater than 200 μmol/l (n=3, Student's t-test, mean±S.D., *P<0.05). (c) Expression of caspase 3 (cleaved-caspase 3, pro-caspase 3) in MMH-R and MMH-RT cells was detected by western blotting (n=3, Student's t-test, mean±S.D., *P<0.05, **P<0.01, ImageJ2X software)
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3973203&req=5

fig5: Apoptosis of MMH-R and MMH-RT induced by ATM was determined by flow cytometry. At 48-h ATM incubation in a range of 1–600 μmol/l, the apoptotic rates of MMH-R and MMH-RT cells were detected by flow cytometry. (a) Typical FACS schematic. (b) Following the increased ATM concentration, the standardized apoptotic rates (%) of mesenchymal MMH-RT cells were dramatically higher than those of epithelial MMH-R cells when the concentration of ATM treatment was greater than 200 μmol/l (n=3, Student's t-test, mean±S.D., *P<0.05). (c) Expression of caspase 3 (cleaved-caspase 3, pro-caspase 3) in MMH-R and MMH-RT cells was detected by western blotting (n=3, Student's t-test, mean±S.D., *P<0.05, **P<0.01, ImageJ2X software)
Mentions: ATM-induced apoptosis in 48 h was detected by fluorescence activated cell sorter (FACS) (Figure 5a). The results showed that the standardized apoptotic rate (%) of mesenchymal MMH-RT cells was dramatically higher than that of epithelial MMH-R cells when concentration of ATM treatment was above 200 μmol/l (Figure 5b, Student's t-test, P<0.05, Supplementary Figure 2), while in MMH-R cells, the apoptosis rate had no obvious correlation with concentration (Figure 5b; Supplementary Figure 2). We also detected the expression of caspase 3, a key molecule of apoptosis signal pathway, in the MMH-R and MMH-RT cells. The results exhibited that the expression of cleaved-caspase 3 of MMH-RT cells was significantly higher than that of MMH-R cells with the increased concentration of ATM (Figure 5c, Student's t-test, P<0.01). These findings indicated that the aPKCι signaling inhibitor could activate the caspase 3-dependent apoptosis pathway and significantly promote the apoptosis of MMH-RT cells.

Bottom Line: Our study showed that EMT took place in MMH-R cells under the effect of transforming growth factor-β1 (TGF-β1) overexpressing aPKCι.In conclusion, our result suggested that aPKCι could be an important bio-marker of tumor EMT, and used as an indicator of invasion and malignancy.ATM might be a promising agent for targeted treatment of HCC.

View Article: PubMed Central - PubMed

Affiliation: Department of Biliary and Pancreatic Surgery/Cancer Research Center Affiliated Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.

ABSTRACT
Epithelial-to-mesenchymal transition (EMT) has an important role in invasion and metastasis of hepatocellular carcinoma (HCC). To explore the regulatory mechanism of atypical protein kinase C ι (aPKCι) signaling pathways to HCC development, and find an agent for targeted therapy for HCC, immortalized murine hepatocytes were employed to establish an EMT cell model of HCC, MMH-RT cells. Our study showed that EMT took place in MMH-R cells under the effect of transforming growth factor-β1 (TGF-β1) overexpressing aPKCι. Furthermore, we showed that the aPKCι blocking agent aurothiomalate (ATM) inhibited EMT and decreased invasion of hepatocytes. Moreover, ATM selectively inhibited proliferation of mesenchymal cells and HepG2 cells and induced apoptosis. However, ATM increased proliferation of epithelial cells and had little effect on apoptosis and invasion of epithelial cells. In conclusion, our result suggested that aPKCι could be an important bio-marker of tumor EMT, and used as an indicator of invasion and malignancy. ATM might be a promising agent for targeted treatment of HCC.

Show MeSH
Related in: MedlinePlus