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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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Expression of miR-424-5p in detached hepatocarcinoma cells.(a) HCC cells were plated in poly-HEMA coated and non-coated plates and allowed for growth for 24 h as detached and attached cells. MiRNA expression profiling of these two groups was analyzed and the statistically differently expressed microRNAs were selected. (b)BEL7402, SMMC7721, and HepG2 cells were plated in 6-well plates without or with poly-HEMA as attached and detached group. After 24 h of culture, cells were collected and the expression of miR-424-5p in BEL7402, SMMC7721 and HepG2 cells was detected by real-time PCR. Histogram represented the relative miR-424-5p expression levels normalized to U6. Data were presented as mean ± s.d. from triplicate analysis, and were the representative data from at least three independent experiments. *P<0.05. **P<0.01.
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f1: Expression of miR-424-5p in detached hepatocarcinoma cells.(a) HCC cells were plated in poly-HEMA coated and non-coated plates and allowed for growth for 24 h as detached and attached cells. MiRNA expression profiling of these two groups was analyzed and the statistically differently expressed microRNAs were selected. (b)BEL7402, SMMC7721, and HepG2 cells were plated in 6-well plates without or with poly-HEMA as attached and detached group. After 24 h of culture, cells were collected and the expression of miR-424-5p in BEL7402, SMMC7721 and HepG2 cells was detected by real-time PCR. Histogram represented the relative miR-424-5p expression levels normalized to U6. Data were presented as mean ± s.d. from triplicate analysis, and were the representative data from at least three independent experiments. *P<0.05. **P<0.01.

Mentions: Our previous studies have shown that anchorage-deprived HCC cells could resist anoikis and acquire more malignant property after detachment910. Although microRNA is reported to play a critical role in the progression of cancer, whether it is involved in the anoikis-resistance of HCC cells is not known and is investigated in this study. In order to identify miRNAs that are potentially involved in the anoikis-resistant process of HCC cells, we employed a miRNA microarray expression profiling, and screened out a series of miRNAs which were differentially expressed between attached and detached HCC cells. Microarray analysis showed that miR-424-5p was among the top down-regulated hits (Fig. 1a), and its significant down-regulation in the anchorage-deprived HCC cells was further validated by quantitative real-time PCR (Fig. 1b).


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)

Expression of miR-424-5p in detached hepatocarcinoma cells.(a) HCC cells were plated in poly-HEMA coated and non-coated plates and allowed for growth for 24 h as detached and attached cells. MiRNA expression profiling of these two groups was analyzed and the statistically differently expressed microRNAs were selected. (b)BEL7402, SMMC7721, and HepG2 cells were plated in 6-well plates without or with poly-HEMA as attached and detached group. After 24 h of culture, cells were collected and the expression of miR-424-5p in BEL7402, SMMC7721 and HepG2 cells was detected by real-time PCR. Histogram represented the relative miR-424-5p expression levels normalized to U6. Data were presented as mean ± s.d. from triplicate analysis, and were the representative data from at least three independent experiments. *P<0.05. **P<0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f1: Expression of miR-424-5p in detached hepatocarcinoma cells.(a) HCC cells were plated in poly-HEMA coated and non-coated plates and allowed for growth for 24 h as detached and attached cells. MiRNA expression profiling of these two groups was analyzed and the statistically differently expressed microRNAs were selected. (b)BEL7402, SMMC7721, and HepG2 cells were plated in 6-well plates without or with poly-HEMA as attached and detached group. After 24 h of culture, cells were collected and the expression of miR-424-5p in BEL7402, SMMC7721 and HepG2 cells was detected by real-time PCR. Histogram represented the relative miR-424-5p expression levels normalized to U6. Data were presented as mean ± s.d. from triplicate analysis, and were the representative data from at least three independent experiments. *P<0.05. **P<0.01.
Mentions: Our previous studies have shown that anchorage-deprived HCC cells could resist anoikis and acquire more malignant property after detachment910. Although microRNA is reported to play a critical role in the progression of cancer, whether it is involved in the anoikis-resistance of HCC cells is not known and is investigated in this study. In order to identify miRNAs that are potentially involved in the anoikis-resistant process of HCC cells, we employed a miRNA microarray expression profiling, and screened out a series of miRNAs which were differentially expressed between attached and detached HCC cells. Microarray analysis showed that miR-424-5p was among the top down-regulated hits (Fig. 1a), and its significant down-regulation in the anchorage-deprived HCC cells was further validated by quantitative real-time PCR (Fig. 1b).

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