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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.


Related in: MedlinePlus

MiR-429 affected EOC cells partially through KIAA0101. a Transwell migration assay and b Transwell invasion assays in SKOV3 cells transfected with either vector control or plasmids encoding KIAA0101. c MTT cell growth rate was performed in SKOV3 cells transfected with either vector control or plasmids encoding KIAA0101, when treated with Cisplatin (8 µM). *p < 0.05 compared with siR-Control transfected cells
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Fig7: MiR-429 affected EOC cells partially through KIAA0101. a Transwell migration assay and b Transwell invasion assays in SKOV3 cells transfected with either vector control or plasmids encoding KIAA0101. c MTT cell growth rate was performed in SKOV3 cells transfected with either vector control or plasmids encoding KIAA0101, when treated with Cisplatin (8 µM). *p < 0.05 compared with siR-Control transfected cells

Mentions: Our previous results showed that downregulation of KIAA0101 inhibited EOC cell metastasis and Cisplatin chemoresistance, KIAA0101 regulates Wnt/β-catenin signaling and KIAA0101 was a target of miR-429. It is possible that miR-429 exerted its functions on EOC cells via KIAA0101 mediated-Wnt/β-catenin signaling pathway. If this is the case, then the relief of KIAA0101 suppression would reverse the regulation of miR-429 on EOC cells. To test this hypothesis, we simultaneously co-transfected plasmids encoding KIAA0101 into miR-429 overexpressing SKOV3 cells. These plasmids did not contain the 3′-UTR region of KIAA0101, and were therefore resistant to miR-429 regulation. We found that while overexpression of miR-429 reduced the expression of KIAA0101, but co-transfection of KIAA0101-overexpressing plasmids completely restored the levels of KIAA0101 protein as determined by Western blot analysis (Fig. 6a). In these cells, nuclear β-catenin levels and TOP flash luciferase activity inhibited by miR-429 overexpression were partially restored by KIAA0101 overexpression (Fig. 6b, c). The decreased expression of Axin2 and c-myc induced by miR-429 overexpression was partially abrogated by KIAA0101 overexpression (Fig. 6d, e). We then performed cell migration and invasion assays. As shown in Fig. 7a, b, transfection of KIAA0101-expressing plasmid partially reversed the inhibition of EOC cell metastasis (Fig. 7a, b). Furthermore, reduction of cell growth rate induced by miR-429 with Cisplatin treatment was partially restored by KIAA0101 (Fig. 7c). Together, these data suggested that miR-429 inhibited metastasis and chemoresistance of EOC cells, at least in part, via KIAA0101 mediated-Wnt/β-catenin signaling pathway.Fig. 6


KIAA0101, a target gene of miR-429, enhances migration and chemoresistance of epithelial ovarian cancer cells
MiR-429 affected EOC cells partially through KIAA0101. a Transwell migration assay and b Transwell invasion assays in SKOV3 cells transfected with either vector control or plasmids encoding KIAA0101. c MTT cell growth rate was performed in SKOV3 cells transfected with either vector control or plasmids encoding KIAA0101, when treated with Cisplatin (8 µM). *p < 0.05 compared with siR-Control transfected cells
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Related In: Results  -  Collection

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Fig7: MiR-429 affected EOC cells partially through KIAA0101. a Transwell migration assay and b Transwell invasion assays in SKOV3 cells transfected with either vector control or plasmids encoding KIAA0101. c MTT cell growth rate was performed in SKOV3 cells transfected with either vector control or plasmids encoding KIAA0101, when treated with Cisplatin (8 µM). *p < 0.05 compared with siR-Control transfected cells
Mentions: Our previous results showed that downregulation of KIAA0101 inhibited EOC cell metastasis and Cisplatin chemoresistance, KIAA0101 regulates Wnt/β-catenin signaling and KIAA0101 was a target of miR-429. It is possible that miR-429 exerted its functions on EOC cells via KIAA0101 mediated-Wnt/β-catenin signaling pathway. If this is the case, then the relief of KIAA0101 suppression would reverse the regulation of miR-429 on EOC cells. To test this hypothesis, we simultaneously co-transfected plasmids encoding KIAA0101 into miR-429 overexpressing SKOV3 cells. These plasmids did not contain the 3′-UTR region of KIAA0101, and were therefore resistant to miR-429 regulation. We found that while overexpression of miR-429 reduced the expression of KIAA0101, but co-transfection of KIAA0101-overexpressing plasmids completely restored the levels of KIAA0101 protein as determined by Western blot analysis (Fig. 6a). In these cells, nuclear β-catenin levels and TOP flash luciferase activity inhibited by miR-429 overexpression were partially restored by KIAA0101 overexpression (Fig. 6b, c). The decreased expression of Axin2 and c-myc induced by miR-429 overexpression was partially abrogated by KIAA0101 overexpression (Fig. 6d, e). We then performed cell migration and invasion assays. As shown in Fig. 7a, b, transfection of KIAA0101-expressing plasmid partially reversed the inhibition of EOC cell metastasis (Fig. 7a, b). Furthermore, reduction of cell growth rate induced by miR-429 with Cisplatin treatment was partially restored by KIAA0101 (Fig. 7c). Together, these data suggested that miR-429 inhibited metastasis and chemoresistance of EOC cells, at least in part, via KIAA0101 mediated-Wnt/β-catenin signaling pathway.Fig. 6

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.


Related in: MedlinePlus