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KIAA0101, a target gene of miR-429, enhances migration and chemoresistance of epithelial ovarian cancer cells

View Article: PubMed Central - PubMed

ABSTRACT

Background: Ovarian cancer is a common type of gynecological malignancies, and is the fifth leading cause of cancer-related death in women in the United States. MiR-429 and KIAA0101 have been found to be involved in several human malignancies, respectively. However, the role of miR-429 and KIAA0101, and the correlation between them during development of epithelial ovarian cancer (EOC) remain to be investigated.

Methods: The expression of KIAA0101 in EOC tissues and cells was measured by Quantitative real-time PCR, western blot, and immunochemistry. Cell proliferation assay, colony formation assay, and transwell assay was performed to assess the role of miR-429 and KIAA0101 in regulation of proliferation, migration, and chemoresistance of EOC cells. Luciferase assay was used to test the Wnt/β-catenin signaling activity in response to depletion of KIAA0101 and overexpression of miR-429.

Results: We found that KIAA0101 was upregulated in metastatic EOC tissues, compared to primary EOC tissues, and KIAA0101 was required for the migration activity and chemoresistance of EOC cells by enhancing Wnt/β-catenin signaling. Furthermore, we revealed KIAA0101 is direct target of miR-429. Similar to knockdown of KIAA0101, overexpression of miR-429 reduced invasion and chemoresistance of EOC cells. Co-transfection of KIAA0101 partially abrogates the inhibitory effects on invasion and chemoresistance in EOC cells.

Conclusions: KIAA0101, a target gene of miR-429, was upregulated in the metastatic EOC tissues, and enhanced the migration activity and chemoresistance of EOC cells. Both miR-429 and KIAA0101 may represent the potential therapeutic targets of EOC.

No MeSH data available.


KIAA0101 is targeted by miR-429. a The putative binding sites of miR-429 on the KIAA0101 3′-UTR region. b Wild type (WT) of KIAA01013′-UTR region was mutated. c Luciferase activities in SKOV3 and COV644 cells 48 h after co-transfected with KIAA0101 3′-UTR wild type or mutant (Mut) luciferase plasmid and miR-429 or miR-Control. **p < 0.01 compared to miR-Control transfected cells. d Levels of KIAA0101 protein was determined by Western blot in SKOV3 and COV644 cells transfected with miR-429 or miR-Control. e The correlation of the expression levels of KIAA0101 and miR-429 in 40 EOC tissue samples (r = −0.648, p < 0.001)
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Fig5: KIAA0101 is targeted by miR-429. a The putative binding sites of miR-429 on the KIAA0101 3′-UTR region. b Wild type (WT) of KIAA01013′-UTR region was mutated. c Luciferase activities in SKOV3 and COV644 cells 48 h after co-transfected with KIAA0101 3′-UTR wild type or mutant (Mut) luciferase plasmid and miR-429 or miR-Control. **p < 0.01 compared to miR-Control transfected cells. d Levels of KIAA0101 protein was determined by Western blot in SKOV3 and COV644 cells transfected with miR-429 or miR-Control. e The correlation of the expression levels of KIAA0101 and miR-429 in 40 EOC tissue samples (r = −0.648, p < 0.001)

Mentions: We employed the bioinformatic Target Scan tools to predict the miRNA targeting KIAA0101, and found that miR-429 was one of the miRNA potentially binding KIAA0101 (Fig. 5a). MiR-429 plays an important role in EOC metastasis and chemoresistance. In order to validate that KIAA0101 was a direct target gene of miR-429, we constructed luciferase reporter plasmid with the wild type and mutant KIAA0101 3′-UTR region (Fig. 5b). We then co-transfected these plasmids into HEK293T cells with miR-429 and control miRNA, respectively. Results showed the transfection of miR-429 significantly reduced the luciferase activity of wild type KIAA0101 3′-UTR reporter, but not mutant KIAA0101 3′-UTR reporter (Fig. 5c). Consistently, miR-429 remarkably reduced KIAA0101 levels in SKOV3 and COV644 cells (Fig. 5d). Furthermore, we found an inverse correlation between the expression of KIAA0101 and miR-429 in 40 EOC tissue samples (Fig. 5e). Collectively, these results indicated that miR-429 inhibited the expression of KIAA0101 by targeting its 3′-UTR region.Fig. 5


KIAA0101, a target gene of miR-429, enhances migration and chemoresistance of epithelial ovarian cancer cells
KIAA0101 is targeted by miR-429. a The putative binding sites of miR-429 on the KIAA0101 3′-UTR region. b Wild type (WT) of KIAA01013′-UTR region was mutated. c Luciferase activities in SKOV3 and COV644 cells 48 h after co-transfected with KIAA0101 3′-UTR wild type or mutant (Mut) luciferase plasmid and miR-429 or miR-Control. **p < 0.01 compared to miR-Control transfected cells. d Levels of KIAA0101 protein was determined by Western blot in SKOV3 and COV644 cells transfected with miR-429 or miR-Control. e The correlation of the expression levels of KIAA0101 and miR-429 in 40 EOC tissue samples (r = −0.648, p < 0.001)
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Fig5: KIAA0101 is targeted by miR-429. a The putative binding sites of miR-429 on the KIAA0101 3′-UTR region. b Wild type (WT) of KIAA01013′-UTR region was mutated. c Luciferase activities in SKOV3 and COV644 cells 48 h after co-transfected with KIAA0101 3′-UTR wild type or mutant (Mut) luciferase plasmid and miR-429 or miR-Control. **p < 0.01 compared to miR-Control transfected cells. d Levels of KIAA0101 protein was determined by Western blot in SKOV3 and COV644 cells transfected with miR-429 or miR-Control. e The correlation of the expression levels of KIAA0101 and miR-429 in 40 EOC tissue samples (r = −0.648, p < 0.001)
Mentions: We employed the bioinformatic Target Scan tools to predict the miRNA targeting KIAA0101, and found that miR-429 was one of the miRNA potentially binding KIAA0101 (Fig. 5a). MiR-429 plays an important role in EOC metastasis and chemoresistance. In order to validate that KIAA0101 was a direct target gene of miR-429, we constructed luciferase reporter plasmid with the wild type and mutant KIAA0101 3′-UTR region (Fig. 5b). We then co-transfected these plasmids into HEK293T cells with miR-429 and control miRNA, respectively. Results showed the transfection of miR-429 significantly reduced the luciferase activity of wild type KIAA0101 3′-UTR reporter, but not mutant KIAA0101 3′-UTR reporter (Fig. 5c). Consistently, miR-429 remarkably reduced KIAA0101 levels in SKOV3 and COV644 cells (Fig. 5d). Furthermore, we found an inverse correlation between the expression of KIAA0101 and miR-429 in 40 EOC tissue samples (Fig. 5e). Collectively, these results indicated that miR-429 inhibited the expression of KIAA0101 by targeting its 3′-UTR region.Fig. 5

View Article: PubMed Central - PubMed

ABSTRACT

Background: Ovarian cancer is a common type of gynecological malignancies, and is the fifth leading cause of cancer-related death in women in the United States. MiR-429 and KIAA0101 have been found to be involved in several human malignancies, respectively. However, the role of miR-429 and KIAA0101, and the correlation between them during development of epithelial ovarian cancer (EOC) remain to be investigated.

Methods: The expression of KIAA0101 in EOC tissues and cells was measured by Quantitative real-time PCR, western blot, and immunochemistry. Cell proliferation assay, colony formation assay, and transwell assay was performed to assess the role of miR-429 and KIAA0101 in regulation of proliferation, migration, and chemoresistance of EOC cells. Luciferase assay was used to test the Wnt/&beta;-catenin signaling activity in response to depletion of KIAA0101 and overexpression of miR-429.

Results: We found that KIAA0101 was upregulated in metastatic EOC tissues, compared to primary EOC tissues, and KIAA0101 was required for the migration activity and chemoresistance of EOC cells by enhancing Wnt/&beta;-catenin signaling. Furthermore, we revealed KIAA0101 is direct target of miR-429. Similar to knockdown of KIAA0101, overexpression of miR-429 reduced invasion and chemoresistance of EOC cells. Co-transfection of KIAA0101 partially abrogates the inhibitory effects on invasion and chemoresistance in EOC cells.

Conclusions: KIAA0101, a target gene of miR-429, was upregulated in the metastatic EOC tissues, and enhanced the migration activity and chemoresistance of EOC cells. Both miR-429 and KIAA0101 may represent the potential therapeutic targets of EOC.

No MeSH data available.