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MicroRNA-200a suppresses metastatic potential of side population cells in human hepatocellular carcinoma by decreasing ZEB2.

Yang X, Wang J, Qu S, Zhang H, Ruan B, Gao Y, Ma B, Wang X, Wu N, Li X, Dou K, Li H - Oncotarget (2015)

Bottom Line: We found that miR-200a was downregulated in HCC/SP and this was associated metastasis.Overexpression of miR-200a in SP cells decreased metastasis-related markers and expression of ZEB2.The associations between miR-200a, SP cells and ZEB2 were validated in HCC.

View Article: PubMed Central - PubMed

Affiliation: Department of Hepatobiliary Surgery, The Xijing Hospital of The Fourth Military Medical Uiversity, Xi'an, China.

ABSTRACT
Although microRNA-200a (miR-200a) is frequently downregulated in cancer, its role in side population (SP) has not been investigated. In this study, 101 pairs of primary hepatocellular carcinoma (HCC) tissues and matched normal control tissues were analyzed for miR-200a expression and its clinicopathological value was determined. We found that miR-200a was downregulated in HCC/SP and this was associated metastasis. MiR-200a suppressed metastasis of SP cells. Overexpression of miR-200a in SP cells decreased metastasis-related markers and expression of ZEB2. The associations between miR-200a, SP cells and ZEB2 were validated in HCC. These findings reveal that miR-200a suppresses metastasis of SP cells by downregulating ZEB2.

No MeSH data available.


Related in: MedlinePlus

MiR-200a induces the metastasis of SP cells through the transactivation of ZEB2 expression(A) The miR-200a target site in the 3′UTR of ZEB2. (B) The up-regulation or down-regulation of miR-200a in MHCC-97H demonstrated an effect on ZEB2 according to western-blot. (C) Luciferase activity of ZEB2 after transfection with miR-200a mimics or an inhibitor. (D) Functional evaluation of miR-200a induction on its validated target, ZEB, in different groups by qRT-PCR. (E) Real-time PCR and (F) western-blot were used to detect the expression of EMT markers. (G) Following the infection of the MHCC-97H-SP-miR-200a cells and Huh7-SP-KD-miR-200a cells with ZEB2 siRNA and the designated SP-ZEB2 and SP-anti-ZEB2. The cell invasion and migration capacities were assessed with a transwell assay. *p < 0.05, **p < 0.01, t test.
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Figure 7: MiR-200a induces the metastasis of SP cells through the transactivation of ZEB2 expression(A) The miR-200a target site in the 3′UTR of ZEB2. (B) The up-regulation or down-regulation of miR-200a in MHCC-97H demonstrated an effect on ZEB2 according to western-blot. (C) Luciferase activity of ZEB2 after transfection with miR-200a mimics or an inhibitor. (D) Functional evaluation of miR-200a induction on its validated target, ZEB, in different groups by qRT-PCR. (E) Real-time PCR and (F) western-blot were used to detect the expression of EMT markers. (G) Following the infection of the MHCC-97H-SP-miR-200a cells and Huh7-SP-KD-miR-200a cells with ZEB2 siRNA and the designated SP-ZEB2 and SP-anti-ZEB2. The cell invasion and migration capacities were assessed with a transwell assay. *p < 0.05, **p < 0.01, t test.

Mentions: The target site of miR-200a in the 3′UTR of ZEB2 was predicted (Fig. 7A). We also over-expressed or downregulated the miR-200a expression in MHCC-97H, and by western-blot we determined that miR-200a had the opposite function of ZEB2 (Fig. 7B). A luciferase reporter gene assay showed that, compared with the NC group, miR-200a mimics significantly inhibited the 3′UTR wild-type (WT) ZEB2 activity (P < 0.01) and a miR-200a inhibitor significantly increased its activity (P < 0.01). MiR-200a mimics and a miR-200a inhibitor had no effect on the activity of 3′UTR mutant (MT) ZEB2 (Fig. 7C) (Y-axis signifies the luciferase activity of ZEB2).


MicroRNA-200a suppresses metastatic potential of side population cells in human hepatocellular carcinoma by decreasing ZEB2.

Yang X, Wang J, Qu S, Zhang H, Ruan B, Gao Y, Ma B, Wang X, Wu N, Li X, Dou K, Li H - Oncotarget (2015)

MiR-200a induces the metastasis of SP cells through the transactivation of ZEB2 expression(A) The miR-200a target site in the 3′UTR of ZEB2. (B) The up-regulation or down-regulation of miR-200a in MHCC-97H demonstrated an effect on ZEB2 according to western-blot. (C) Luciferase activity of ZEB2 after transfection with miR-200a mimics or an inhibitor. (D) Functional evaluation of miR-200a induction on its validated target, ZEB, in different groups by qRT-PCR. (E) Real-time PCR and (F) western-blot were used to detect the expression of EMT markers. (G) Following the infection of the MHCC-97H-SP-miR-200a cells and Huh7-SP-KD-miR-200a cells with ZEB2 siRNA and the designated SP-ZEB2 and SP-anti-ZEB2. The cell invasion and migration capacities were assessed with a transwell assay. *p < 0.05, **p < 0.01, t test.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4480725&req=5

Figure 7: MiR-200a induces the metastasis of SP cells through the transactivation of ZEB2 expression(A) The miR-200a target site in the 3′UTR of ZEB2. (B) The up-regulation or down-regulation of miR-200a in MHCC-97H demonstrated an effect on ZEB2 according to western-blot. (C) Luciferase activity of ZEB2 after transfection with miR-200a mimics or an inhibitor. (D) Functional evaluation of miR-200a induction on its validated target, ZEB, in different groups by qRT-PCR. (E) Real-time PCR and (F) western-blot were used to detect the expression of EMT markers. (G) Following the infection of the MHCC-97H-SP-miR-200a cells and Huh7-SP-KD-miR-200a cells with ZEB2 siRNA and the designated SP-ZEB2 and SP-anti-ZEB2. The cell invasion and migration capacities were assessed with a transwell assay. *p < 0.05, **p < 0.01, t test.
Mentions: The target site of miR-200a in the 3′UTR of ZEB2 was predicted (Fig. 7A). We also over-expressed or downregulated the miR-200a expression in MHCC-97H, and by western-blot we determined that miR-200a had the opposite function of ZEB2 (Fig. 7B). A luciferase reporter gene assay showed that, compared with the NC group, miR-200a mimics significantly inhibited the 3′UTR wild-type (WT) ZEB2 activity (P < 0.01) and a miR-200a inhibitor significantly increased its activity (P < 0.01). MiR-200a mimics and a miR-200a inhibitor had no effect on the activity of 3′UTR mutant (MT) ZEB2 (Fig. 7C) (Y-axis signifies the luciferase activity of ZEB2).

Bottom Line: We found that miR-200a was downregulated in HCC/SP and this was associated metastasis.Overexpression of miR-200a in SP cells decreased metastasis-related markers and expression of ZEB2.The associations between miR-200a, SP cells and ZEB2 were validated in HCC.

View Article: PubMed Central - PubMed

Affiliation: Department of Hepatobiliary Surgery, The Xijing Hospital of The Fourth Military Medical Uiversity, Xi'an, China.

ABSTRACT
Although microRNA-200a (miR-200a) is frequently downregulated in cancer, its role in side population (SP) has not been investigated. In this study, 101 pairs of primary hepatocellular carcinoma (HCC) tissues and matched normal control tissues were analyzed for miR-200a expression and its clinicopathological value was determined. We found that miR-200a was downregulated in HCC/SP and this was associated metastasis. MiR-200a suppressed metastasis of SP cells. Overexpression of miR-200a in SP cells decreased metastasis-related markers and expression of ZEB2. The associations between miR-200a, SP cells and ZEB2 were validated in HCC. These findings reveal that miR-200a suppresses metastasis of SP cells by downregulating ZEB2.

No MeSH data available.


Related in: MedlinePlus