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MiR-223-3p targeting SEPT6 promotes the biological behavior of prostate cancer.

Wei Y, Yang J, Yi L, Wang Y, Dong Z, Liu Z, Ou-yang S, Wu H, Zhong Z, Yin Z, Zhou K, Gao Y, Yan B, Wang Z - Sci Rep (2014)

Bottom Line: However, whether altered expression of miR-223 is associated with prostate cancer (PCa) and what it is potential functions in PCa remained unveiled.In this study, we firstly found miR-223-3p were up-regulated in prostate cancer tissues and then we study functional role of miR-223-3p in PCa using DU145, PC3 and LNCaP cell lines.Notably, we found increasing SEPT6 expression might reverse the biological activity induced by miR-223-3p, which might be a potential therapeutic target for PCa.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, the Second Xiangya Hospital, Central South University, Changsha 410011, China.

ABSTRACT
MicroRNAs (miRNAs) present frequently altered expression in urologic cancers including prostate, bladder, and kidney cancer. The altered expression of miR-223 has been reported in cancers and other diseases in recent researches. MiR-223 is up-regulated in systemic lupus erythematosus and rheumatoid arthritis. In neoplastic diseases, miR-223 is proved to be up-expressed in plasma or serum and cancer tissues compared with normal tissues in pancreatic cancer, gastric cancer, et al. However, whether altered expression of miR-223 is associated with prostate cancer (PCa) and what it is potential functions in PCa remained unveiled. In this study, we firstly found miR-223-3p were up-regulated in prostate cancer tissues and then we study functional role of miR-223-3p in PCa using DU145, PC3 and LNCaP cell lines. Our data suggested that miR-223-3p might target gene SEPT6 and promoted the biological behavior of prostate cancer. Notably, we found increasing SEPT6 expression might reverse the biological activity induced by miR-223-3p, which might be a potential therapeutic target for PCa.

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Related in: MedlinePlus

MiR-223-3p regulates biological behavior in PCa cell lines.Figure 2A1 and 2A2: Fluorescence microscope is used for detecting transfection efficiency for miR-MM and miR-AO transfection and the results suggested transfection efficiencies are all more than 90%. Figure 2B: MiR-223-3p expression is evaluated in groups after the different transfections, resulting in different expression levels of miR-223-3p are existed among four groups in each cell lines. MiR-223-3p contents in miR-MM groups are significantly higher than the other three groups (all p < 0.01), while expression in groups of miR-AO are lowest (all p < 0.01). Figure 2C1, 2C2 and 2C3: cell proliferation assays for DU145, PC3 and LNCaP, show that knocking down miR-223-3p expression (groups of miR-AO) leads to lowest cell proliferation, cells with up-regulated expression of miR-223-3p after miR-MM transfected present highest cell proliferation. Figure 2D, 2E1, 2E2 and 2E3: cell cycle assays for DU145, PC3 and LNCaP. Knocking down miR-223-3p expression makes cells arrest in G0/G1 sub phase, and induces their apoptosis, with the proliferation index (PI) significantly lower than other groups. Figure 2F and 2G: Cell apoptosis assay. The results indicat cells with miR-223-3p knocked down present highest level of apoptosis, while cells with up-regulated expression of miR-223-3p after miR-MM transfected present lowest apoptosis (all p < 0.01). Figure 2H and 2I: In cell invasion assay, cells showed lowest invasive ability in groups of miR-AO, while in contrast, cells with up-regulated expression of miR-223-3p after miR-MM transfected present highest invasive ability (all p < 0.05).** on behalf of p < 0.01. * on behalf of p < 0.05.
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f2: MiR-223-3p regulates biological behavior in PCa cell lines.Figure 2A1 and 2A2: Fluorescence microscope is used for detecting transfection efficiency for miR-MM and miR-AO transfection and the results suggested transfection efficiencies are all more than 90%. Figure 2B: MiR-223-3p expression is evaluated in groups after the different transfections, resulting in different expression levels of miR-223-3p are existed among four groups in each cell lines. MiR-223-3p contents in miR-MM groups are significantly higher than the other three groups (all p < 0.01), while expression in groups of miR-AO are lowest (all p < 0.01). Figure 2C1, 2C2 and 2C3: cell proliferation assays for DU145, PC3 and LNCaP, show that knocking down miR-223-3p expression (groups of miR-AO) leads to lowest cell proliferation, cells with up-regulated expression of miR-223-3p after miR-MM transfected present highest cell proliferation. Figure 2D, 2E1, 2E2 and 2E3: cell cycle assays for DU145, PC3 and LNCaP. Knocking down miR-223-3p expression makes cells arrest in G0/G1 sub phase, and induces their apoptosis, with the proliferation index (PI) significantly lower than other groups. Figure 2F and 2G: Cell apoptosis assay. The results indicat cells with miR-223-3p knocked down present highest level of apoptosis, while cells with up-regulated expression of miR-223-3p after miR-MM transfected present lowest apoptosis (all p < 0.01). Figure 2H and 2I: In cell invasion assay, cells showed lowest invasive ability in groups of miR-AO, while in contrast, cells with up-regulated expression of miR-223-3p after miR-MM transfected present highest invasive ability (all p < 0.05).** on behalf of p < 0.01. * on behalf of p < 0.05.

Mentions: Three PCa cell lines DU145, PC3 and LNCaP cells were divided into four groups. According to previous study, miRNA's mimics and miRNA's antisense oligonucleotides were used for alternating miRNA's expression15. To promote miR-223-3p expression, cells were transfected by miR-mimics (Sense: 5′-ugucaguuugucaaauacccca-3′; antisense:5′-ugggguauuugacaaacugaca-3′) (miR-MM). To inhibit expression of miR-223-3p, cells were knocked down by the miR-223-3p antisense oligonucleotides (5′- ugggguauuugacaaacugaca-3′) (miR-AO). Cells transfected with miRNA negative control (miR-NC) and conventional culture cells (CC-cells) were used as two control groups. Detection of the above transfections was performed on light and fluorescence microscope and by quantitative real-time polymerase chain reaction (qRT-PCR). The results of fluorescence microscope indicated transfection efficiencies in all the two groups of miR-MM and miR-AO were both more than 90% (Figure 2A1 and 2A2). QRT-PCR was carried out for miR-223-3p expression evaluation after the different transfections (Figure 2B). The results suggested the different expression levels of miR-223-3p were existed among four groups in each cell lines. MiR-223-3p contents in miR-MM groups were significantly higher than the other three groups (all p < 0.01). While expression in groups of miR-AO were lowest (all p < 0.01), leaving groups of CC-cells and miR-NC in the middle level and with no significant differences found between them. These results of fluorescence microscope and qRT-PCR demonstrated the above transfections were successful and effective.


MiR-223-3p targeting SEPT6 promotes the biological behavior of prostate cancer.

Wei Y, Yang J, Yi L, Wang Y, Dong Z, Liu Z, Ou-yang S, Wu H, Zhong Z, Yin Z, Zhou K, Gao Y, Yan B, Wang Z - Sci Rep (2014)

MiR-223-3p regulates biological behavior in PCa cell lines.Figure 2A1 and 2A2: Fluorescence microscope is used for detecting transfection efficiency for miR-MM and miR-AO transfection and the results suggested transfection efficiencies are all more than 90%. Figure 2B: MiR-223-3p expression is evaluated in groups after the different transfections, resulting in different expression levels of miR-223-3p are existed among four groups in each cell lines. MiR-223-3p contents in miR-MM groups are significantly higher than the other three groups (all p < 0.01), while expression in groups of miR-AO are lowest (all p < 0.01). Figure 2C1, 2C2 and 2C3: cell proliferation assays for DU145, PC3 and LNCaP, show that knocking down miR-223-3p expression (groups of miR-AO) leads to lowest cell proliferation, cells with up-regulated expression of miR-223-3p after miR-MM transfected present highest cell proliferation. Figure 2D, 2E1, 2E2 and 2E3: cell cycle assays for DU145, PC3 and LNCaP. Knocking down miR-223-3p expression makes cells arrest in G0/G1 sub phase, and induces their apoptosis, with the proliferation index (PI) significantly lower than other groups. Figure 2F and 2G: Cell apoptosis assay. The results indicat cells with miR-223-3p knocked down present highest level of apoptosis, while cells with up-regulated expression of miR-223-3p after miR-MM transfected present lowest apoptosis (all p < 0.01). Figure 2H and 2I: In cell invasion assay, cells showed lowest invasive ability in groups of miR-AO, while in contrast, cells with up-regulated expression of miR-223-3p after miR-MM transfected present highest invasive ability (all p < 0.05).** on behalf of p < 0.01. * on behalf of p < 0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

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f2: MiR-223-3p regulates biological behavior in PCa cell lines.Figure 2A1 and 2A2: Fluorescence microscope is used for detecting transfection efficiency for miR-MM and miR-AO transfection and the results suggested transfection efficiencies are all more than 90%. Figure 2B: MiR-223-3p expression is evaluated in groups after the different transfections, resulting in different expression levels of miR-223-3p are existed among four groups in each cell lines. MiR-223-3p contents in miR-MM groups are significantly higher than the other three groups (all p < 0.01), while expression in groups of miR-AO are lowest (all p < 0.01). Figure 2C1, 2C2 and 2C3: cell proliferation assays for DU145, PC3 and LNCaP, show that knocking down miR-223-3p expression (groups of miR-AO) leads to lowest cell proliferation, cells with up-regulated expression of miR-223-3p after miR-MM transfected present highest cell proliferation. Figure 2D, 2E1, 2E2 and 2E3: cell cycle assays for DU145, PC3 and LNCaP. Knocking down miR-223-3p expression makes cells arrest in G0/G1 sub phase, and induces their apoptosis, with the proliferation index (PI) significantly lower than other groups. Figure 2F and 2G: Cell apoptosis assay. The results indicat cells with miR-223-3p knocked down present highest level of apoptosis, while cells with up-regulated expression of miR-223-3p after miR-MM transfected present lowest apoptosis (all p < 0.01). Figure 2H and 2I: In cell invasion assay, cells showed lowest invasive ability in groups of miR-AO, while in contrast, cells with up-regulated expression of miR-223-3p after miR-MM transfected present highest invasive ability (all p < 0.05).** on behalf of p < 0.01. * on behalf of p < 0.05.
Mentions: Three PCa cell lines DU145, PC3 and LNCaP cells were divided into four groups. According to previous study, miRNA's mimics and miRNA's antisense oligonucleotides were used for alternating miRNA's expression15. To promote miR-223-3p expression, cells were transfected by miR-mimics (Sense: 5′-ugucaguuugucaaauacccca-3′; antisense:5′-ugggguauuugacaaacugaca-3′) (miR-MM). To inhibit expression of miR-223-3p, cells were knocked down by the miR-223-3p antisense oligonucleotides (5′- ugggguauuugacaaacugaca-3′) (miR-AO). Cells transfected with miRNA negative control (miR-NC) and conventional culture cells (CC-cells) were used as two control groups. Detection of the above transfections was performed on light and fluorescence microscope and by quantitative real-time polymerase chain reaction (qRT-PCR). The results of fluorescence microscope indicated transfection efficiencies in all the two groups of miR-MM and miR-AO were both more than 90% (Figure 2A1 and 2A2). QRT-PCR was carried out for miR-223-3p expression evaluation after the different transfections (Figure 2B). The results suggested the different expression levels of miR-223-3p were existed among four groups in each cell lines. MiR-223-3p contents in miR-MM groups were significantly higher than the other three groups (all p < 0.01). While expression in groups of miR-AO were lowest (all p < 0.01), leaving groups of CC-cells and miR-NC in the middle level and with no significant differences found between them. These results of fluorescence microscope and qRT-PCR demonstrated the above transfections were successful and effective.

Bottom Line: However, whether altered expression of miR-223 is associated with prostate cancer (PCa) and what it is potential functions in PCa remained unveiled.In this study, we firstly found miR-223-3p were up-regulated in prostate cancer tissues and then we study functional role of miR-223-3p in PCa using DU145, PC3 and LNCaP cell lines.Notably, we found increasing SEPT6 expression might reverse the biological activity induced by miR-223-3p, which might be a potential therapeutic target for PCa.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, the Second Xiangya Hospital, Central South University, Changsha 410011, China.

ABSTRACT
MicroRNAs (miRNAs) present frequently altered expression in urologic cancers including prostate, bladder, and kidney cancer. The altered expression of miR-223 has been reported in cancers and other diseases in recent researches. MiR-223 is up-regulated in systemic lupus erythematosus and rheumatoid arthritis. In neoplastic diseases, miR-223 is proved to be up-expressed in plasma or serum and cancer tissues compared with normal tissues in pancreatic cancer, gastric cancer, et al. However, whether altered expression of miR-223 is associated with prostate cancer (PCa) and what it is potential functions in PCa remained unveiled. In this study, we firstly found miR-223-3p were up-regulated in prostate cancer tissues and then we study functional role of miR-223-3p in PCa using DU145, PC3 and LNCaP cell lines. Our data suggested that miR-223-3p might target gene SEPT6 and promoted the biological behavior of prostate cancer. Notably, we found increasing SEPT6 expression might reverse the biological activity induced by miR-223-3p, which might be a potential therapeutic target for PCa.

Show MeSH
Related in: MedlinePlus