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MiR-424-5p reversed epithelial-mesenchymal transition of anchorage-independent HCC cells by directly targeting ICAT and suppressed HCC progression.

Zhang Y, Li T, Guo P, Kang J, Wei Q, Jia X, Zhao W, Huai W, Qiu Y, Sun L, Han L - Sci Rep (2014)

Bottom Line: Microarray expression profiling revealed that expression of miR-424-5p was significantly decreased in anoikis-resistant HCC cells.Clinical investigation demonstrated that miR-424-5p was significantly downregulated in HCC tissues compared with that of the non-cancerous liver tissues, and this decreased expression of miR-424-5p was significantly correlated with higher pathological grades and more advanced TNM stages.Therefore, aberrant expression of miR-424-5p is critically involved in resistance to anoikis and EMT during the metastatic process of HCC, and its downregulation significantly contributes to liver cancer progression.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, Shandong University School of Medicine, Jinan 250012, China.

ABSTRACT
Resistance to anoikis and Epithelial-mesenchymal transition (EMT) are two processes critically involved in cancer metastasis. In this study, we demonstrated that after anchorage deprival, hepatocellular carcinoma (HCC) cells not only resisted anoikis, but also exhibited EMT process. Microarray expression profiling revealed that expression of miR-424-5p was significantly decreased in anoikis-resistant HCC cells. Ectopic overexpression of miR-424-5p was sufficient to reverse resistance to anoikis, block EMT process and inhibit malignant behaviors of HCC cells. Target analysis showed that a potent β-catenin inhibitor, ICAT/CTNNBIP1 was a direct target of miR-424-5p. Further study demonstrated that miR-424-5p reversed resistance to anoikis and EMT of HCCs by directly targeting ICAT and further maintaining the E-cadherin/β-catanin complex on the cellular membrance. In vivo study further demonstrated that miR-424-5p significantly inhibited the tumorigenicity of HCC cells in nude mice. Clinical investigation demonstrated that miR-424-5p was significantly downregulated in HCC tissues compared with that of the non-cancerous liver tissues, and this decreased expression of miR-424-5p was significantly correlated with higher pathological grades and more advanced TNM stages. Therefore, aberrant expression of miR-424-5p is critically involved in resistance to anoikis and EMT during the metastatic process of HCC, and its downregulation significantly contributes to liver cancer progression.

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Enforced miR-424-5p expression reversed EMT-like features and malignant behaviors of HCC cells.(a) HepG2 cells were transfected with miR-424-5p or miR-NC and cultured in anchorage deprived condition for 24 h before expression analysis of the EMT related markers. (b) After cultured in anchorage deprived condition for 24 h, miR-424-5p transfected HCC cells were seeded into normal 6-well plate at the density of 1,000 cells/well and incubated for 7 days. The resulting colonies were stained with crystal violet and counted. (c) MiR-424-5p transfected BEL7402, SMMC7721 and HepG2 cells were cultured in anchorage deprived condition for 24 h before invasion activities were measured. Photos were representative fields of invasive cells on the membrane. All of the presented data are representative one from at least three independent experiments. *P < 0.05,**P < 0.01, *** P < 0.001.
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f4: Enforced miR-424-5p expression reversed EMT-like features and malignant behaviors of HCC cells.(a) HepG2 cells were transfected with miR-424-5p or miR-NC and cultured in anchorage deprived condition for 24 h before expression analysis of the EMT related markers. (b) After cultured in anchorage deprived condition for 24 h, miR-424-5p transfected HCC cells were seeded into normal 6-well plate at the density of 1,000 cells/well and incubated for 7 days. The resulting colonies were stained with crystal violet and counted. (c) MiR-424-5p transfected BEL7402, SMMC7721 and HepG2 cells were cultured in anchorage deprived condition for 24 h before invasion activities were measured. Photos were representative fields of invasive cells on the membrane. All of the presented data are representative one from at least three independent experiments. *P < 0.05,**P < 0.01, *** P < 0.001.

Mentions: Besides conquering resistance to anoikis, ectopic overexpression of miR-424-5p was also accompanied with decreased expression of mesenchymal markers (N-cadherin, Vimentin) and increased expression of epithelial markers(E-cadherin, β-catenin) (Fig. 4a), which indicated a reversion of EMT in the detached HCC cells as well. Furthermore, other malignant behaviors accompanied with resistance to anoikis, including colony formation and invasion was also significantly inhibited by overexpression of miR-424-5p (Fig. 4b–c). Taken together, these data indicated a critical role of miR-424-5p involved in HCC progression by suppressing anoikis and reversing EMT related malignant behaviors of detached HCC cells.


MiR-424-5p reversed epithelial-mesenchymal transition of anchorage-independent HCC cells by directly targeting ICAT and suppressed HCC progression.

Zhang Y, Li T, Guo P, Kang J, Wei Q, Jia X, Zhao W, Huai W, Qiu Y, Sun L, Han L - Sci Rep (2014)

Enforced miR-424-5p expression reversed EMT-like features and malignant behaviors of HCC cells.(a) HepG2 cells were transfected with miR-424-5p or miR-NC and cultured in anchorage deprived condition for 24 h before expression analysis of the EMT related markers. (b) After cultured in anchorage deprived condition for 24 h, miR-424-5p transfected HCC cells were seeded into normal 6-well plate at the density of 1,000 cells/well and incubated for 7 days. The resulting colonies were stained with crystal violet and counted. (c) MiR-424-5p transfected BEL7402, SMMC7721 and HepG2 cells were cultured in anchorage deprived condition for 24 h before invasion activities were measured. Photos were representative fields of invasive cells on the membrane. All of the presented data are representative one from at least three independent experiments. *P < 0.05,**P < 0.01, *** P < 0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4150107&req=5

f4: Enforced miR-424-5p expression reversed EMT-like features and malignant behaviors of HCC cells.(a) HepG2 cells were transfected with miR-424-5p or miR-NC and cultured in anchorage deprived condition for 24 h before expression analysis of the EMT related markers. (b) After cultured in anchorage deprived condition for 24 h, miR-424-5p transfected HCC cells were seeded into normal 6-well plate at the density of 1,000 cells/well and incubated for 7 days. The resulting colonies were stained with crystal violet and counted. (c) MiR-424-5p transfected BEL7402, SMMC7721 and HepG2 cells were cultured in anchorage deprived condition for 24 h before invasion activities were measured. Photos were representative fields of invasive cells on the membrane. All of the presented data are representative one from at least three independent experiments. *P < 0.05,**P < 0.01, *** P < 0.001.
Mentions: Besides conquering resistance to anoikis, ectopic overexpression of miR-424-5p was also accompanied with decreased expression of mesenchymal markers (N-cadherin, Vimentin) and increased expression of epithelial markers(E-cadherin, β-catenin) (Fig. 4a), which indicated a reversion of EMT in the detached HCC cells as well. Furthermore, other malignant behaviors accompanied with resistance to anoikis, including colony formation and invasion was also significantly inhibited by overexpression of miR-424-5p (Fig. 4b–c). Taken together, these data indicated a critical role of miR-424-5p involved in HCC progression by suppressing anoikis and reversing EMT related malignant behaviors of detached HCC cells.

Bottom Line: Microarray expression profiling revealed that expression of miR-424-5p was significantly decreased in anoikis-resistant HCC cells.Clinical investigation demonstrated that miR-424-5p was significantly downregulated in HCC tissues compared with that of the non-cancerous liver tissues, and this decreased expression of miR-424-5p was significantly correlated with higher pathological grades and more advanced TNM stages.Therefore, aberrant expression of miR-424-5p is critically involved in resistance to anoikis and EMT during the metastatic process of HCC, and its downregulation significantly contributes to liver cancer progression.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, Shandong University School of Medicine, Jinan 250012, China.

ABSTRACT
Resistance to anoikis and Epithelial-mesenchymal transition (EMT) are two processes critically involved in cancer metastasis. In this study, we demonstrated that after anchorage deprival, hepatocellular carcinoma (HCC) cells not only resisted anoikis, but also exhibited EMT process. Microarray expression profiling revealed that expression of miR-424-5p was significantly decreased in anoikis-resistant HCC cells. Ectopic overexpression of miR-424-5p was sufficient to reverse resistance to anoikis, block EMT process and inhibit malignant behaviors of HCC cells. Target analysis showed that a potent β-catenin inhibitor, ICAT/CTNNBIP1 was a direct target of miR-424-5p. Further study demonstrated that miR-424-5p reversed resistance to anoikis and EMT of HCCs by directly targeting ICAT and further maintaining the E-cadherin/β-catanin complex on the cellular membrance. In vivo study further demonstrated that miR-424-5p significantly inhibited the tumorigenicity of HCC cells in nude mice. Clinical investigation demonstrated that miR-424-5p was significantly downregulated in HCC tissues compared with that of the non-cancerous liver tissues, and this decreased expression of miR-424-5p was significantly correlated with higher pathological grades and more advanced TNM stages. Therefore, aberrant expression of miR-424-5p is critically involved in resistance to anoikis and EMT during the metastatic process of HCC, and its downregulation significantly contributes to liver cancer progression.

Show MeSH
Related in: MedlinePlus