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Roles of DPY30 in the Proliferation and Motility of Gastric Cancer Cells.

Lee YJ, Han ME, Baek SJ, Kim SY, Oh SO - PLoS ONE (2015)

Bottom Line: Various types of histone methylation have been associated with cancer progression.Depending on the methylation site in histone proteins, its effects on transcription are different.Its knockdown by siRNA decreased the proliferation, migration, and invasion of gastric cancer cells, whereas its overexpression showed the opposite effects.

View Article: PubMed Central - PubMed

Affiliation: Departments of Anatomy, School of Medicine, Pusan National University, Busan, Republic of Korea.

ABSTRACT
Various types of histone methylation have been associated with cancer progression. Depending on the methylation site in histone proteins, its effects on transcription are different. DPY30 is a common member of SET1/MLL histone H3K4 methyltransferase complexes. However, its expression and roles in gastric cancer have been poorly characterized. To determine whether DPY30 has pathophysiological roles in gastric cancer, its expression and roles were examined. Immunohistochemistry and real time PCR showed up-regulation of DPY30 expression in some gastric cancer cell lines and patients' tissues. Its knockdown by siRNA decreased the proliferation, migration, and invasion of gastric cancer cells, whereas its overexpression showed the opposite effects. These results indicate that DPY30 has critical roles in the proliferation, migration, and invasion of gastric cancer cells, and suggest DPY30 might be a therapeutic target in gastric cancer.

No MeSH data available.


Related in: MedlinePlus

DPY30 regulated the proliferation of gastric cancer cells.(A) Real-time PCR was used to determine the efficiency of knockdown or overexpression of DPY30 in HFE145, SNU1, SNU16, SNU216 and SNU638 cells. Knockdown efficiency was determined after transfecting cells with 100 nM DPY30 siRNA targeting the ORF or scrambled siRNA (SCR). Overexpression efficiencies were determined in DPY30-overexpressing and mock cells. (B) Effect of DPY30 knockdown or overexpression on cell proliferation. A cell viability assay was used to measure cell proliferation in the presence of 1% FBS. For the DPY30 knockdown experiments, cell viability assays were performed five days after transfecting 100 nM DPY30 siRNA or SCR. After three days of culture, cell viability assays were performed on DPY30-over and mock cells. (C) Expression analysis of DPY30 after known-down or overexpression by siRNAs or DPY30 ORF respectively. The ORF-targeting or the 3’-UTR-targeting siRNA was used for the knock-down. (D) Exogenous DPY30-ORF rescued the inhibition of proliferation by the 3’-UTR-targeting DPY30 siRNA. Mock or DPY30-overexpressing cells were transduced with two kinds of DPY30 siRNA (3’-UTR-targeting or ORF-targeting), then cell viability assay was examined. Values are the means ± SDs of three independent experiments performed in triplicate. *, p < 0.01 (Student’s t test, versus SCR or Mock).
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pone.0131863.g002: DPY30 regulated the proliferation of gastric cancer cells.(A) Real-time PCR was used to determine the efficiency of knockdown or overexpression of DPY30 in HFE145, SNU1, SNU16, SNU216 and SNU638 cells. Knockdown efficiency was determined after transfecting cells with 100 nM DPY30 siRNA targeting the ORF or scrambled siRNA (SCR). Overexpression efficiencies were determined in DPY30-overexpressing and mock cells. (B) Effect of DPY30 knockdown or overexpression on cell proliferation. A cell viability assay was used to measure cell proliferation in the presence of 1% FBS. For the DPY30 knockdown experiments, cell viability assays were performed five days after transfecting 100 nM DPY30 siRNA or SCR. After three days of culture, cell viability assays were performed on DPY30-over and mock cells. (C) Expression analysis of DPY30 after known-down or overexpression by siRNAs or DPY30 ORF respectively. The ORF-targeting or the 3’-UTR-targeting siRNA was used for the knock-down. (D) Exogenous DPY30-ORF rescued the inhibition of proliferation by the 3’-UTR-targeting DPY30 siRNA. Mock or DPY30-overexpressing cells were transduced with two kinds of DPY30 siRNA (3’-UTR-targeting or ORF-targeting), then cell viability assay was examined. Values are the means ± SDs of three independent experiments performed in triplicate. *, p < 0.01 (Student’s t test, versus SCR or Mock).

Mentions: In order to determine possible roles of DPY30 in gastric cancer cells, we first knocked-down DPY30 using the ORF-targeting DPY30 siRNA and monitored its knockdown efficiency by real-time PCR (Fig 2A). The ORF-targeting DPY30 siRNA (100 nM) decreased the mRNA level of DPY30 in HFE145, SNU1, SNU16, SNU216, and SNU638 cells as compared with scrambled siRNA (SCR) by 78%, 89%, 79%, 76%, and 88%, respectively. Five days after transfecting cells with SCR or the ORF-targeting siRNA, we performed proliferation assays. Knockdown of DPY30 inhibited the proliferations of HFE145, SNU1 and SNU16 as compared with SCR by 31%, 69% and 71% respectively, while the knockdown did not affect the proliferations of SNU216 and SNU638 (Fig 2B), in which the expression level of DPY30 was low (Fig 1E). Next, we produced DPY30-overexpressing cell lines (DPY30-over) using HFE145, SNU216 and SNU638 cells, and then compared DPY30 mRNA levels in mock cells (control) and DPY30-over cells. Real-time PCR results showed that the expression level of DPY30 was 4.2, 3.5 and 3.2 fold higher in DPY30-overexpressing HFE145, SNU216 and SNU638 cells than mock cells, respectively (Fig 2A). DPY30 overexpression enhanced the proliferations of HFE145, SNU216 and SNU638 cells versus mock cells by 1.9, 2.2 and 1.6 fold, respectively (Fig 2B).


Roles of DPY30 in the Proliferation and Motility of Gastric Cancer Cells.

Lee YJ, Han ME, Baek SJ, Kim SY, Oh SO - PLoS ONE (2015)

DPY30 regulated the proliferation of gastric cancer cells.(A) Real-time PCR was used to determine the efficiency of knockdown or overexpression of DPY30 in HFE145, SNU1, SNU16, SNU216 and SNU638 cells. Knockdown efficiency was determined after transfecting cells with 100 nM DPY30 siRNA targeting the ORF or scrambled siRNA (SCR). Overexpression efficiencies were determined in DPY30-overexpressing and mock cells. (B) Effect of DPY30 knockdown or overexpression on cell proliferation. A cell viability assay was used to measure cell proliferation in the presence of 1% FBS. For the DPY30 knockdown experiments, cell viability assays were performed five days after transfecting 100 nM DPY30 siRNA or SCR. After three days of culture, cell viability assays were performed on DPY30-over and mock cells. (C) Expression analysis of DPY30 after known-down or overexpression by siRNAs or DPY30 ORF respectively. The ORF-targeting or the 3’-UTR-targeting siRNA was used for the knock-down. (D) Exogenous DPY30-ORF rescued the inhibition of proliferation by the 3’-UTR-targeting DPY30 siRNA. Mock or DPY30-overexpressing cells were transduced with two kinds of DPY30 siRNA (3’-UTR-targeting or ORF-targeting), then cell viability assay was examined. Values are the means ± SDs of three independent experiments performed in triplicate. *, p < 0.01 (Student’s t test, versus SCR or Mock).
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pone.0131863.g002: DPY30 regulated the proliferation of gastric cancer cells.(A) Real-time PCR was used to determine the efficiency of knockdown or overexpression of DPY30 in HFE145, SNU1, SNU16, SNU216 and SNU638 cells. Knockdown efficiency was determined after transfecting cells with 100 nM DPY30 siRNA targeting the ORF or scrambled siRNA (SCR). Overexpression efficiencies were determined in DPY30-overexpressing and mock cells. (B) Effect of DPY30 knockdown or overexpression on cell proliferation. A cell viability assay was used to measure cell proliferation in the presence of 1% FBS. For the DPY30 knockdown experiments, cell viability assays were performed five days after transfecting 100 nM DPY30 siRNA or SCR. After three days of culture, cell viability assays were performed on DPY30-over and mock cells. (C) Expression analysis of DPY30 after known-down or overexpression by siRNAs or DPY30 ORF respectively. The ORF-targeting or the 3’-UTR-targeting siRNA was used for the knock-down. (D) Exogenous DPY30-ORF rescued the inhibition of proliferation by the 3’-UTR-targeting DPY30 siRNA. Mock or DPY30-overexpressing cells were transduced with two kinds of DPY30 siRNA (3’-UTR-targeting or ORF-targeting), then cell viability assay was examined. Values are the means ± SDs of three independent experiments performed in triplicate. *, p < 0.01 (Student’s t test, versus SCR or Mock).
Mentions: In order to determine possible roles of DPY30 in gastric cancer cells, we first knocked-down DPY30 using the ORF-targeting DPY30 siRNA and monitored its knockdown efficiency by real-time PCR (Fig 2A). The ORF-targeting DPY30 siRNA (100 nM) decreased the mRNA level of DPY30 in HFE145, SNU1, SNU16, SNU216, and SNU638 cells as compared with scrambled siRNA (SCR) by 78%, 89%, 79%, 76%, and 88%, respectively. Five days after transfecting cells with SCR or the ORF-targeting siRNA, we performed proliferation assays. Knockdown of DPY30 inhibited the proliferations of HFE145, SNU1 and SNU16 as compared with SCR by 31%, 69% and 71% respectively, while the knockdown did not affect the proliferations of SNU216 and SNU638 (Fig 2B), in which the expression level of DPY30 was low (Fig 1E). Next, we produced DPY30-overexpressing cell lines (DPY30-over) using HFE145, SNU216 and SNU638 cells, and then compared DPY30 mRNA levels in mock cells (control) and DPY30-over cells. Real-time PCR results showed that the expression level of DPY30 was 4.2, 3.5 and 3.2 fold higher in DPY30-overexpressing HFE145, SNU216 and SNU638 cells than mock cells, respectively (Fig 2A). DPY30 overexpression enhanced the proliferations of HFE145, SNU216 and SNU638 cells versus mock cells by 1.9, 2.2 and 1.6 fold, respectively (Fig 2B).

Bottom Line: Various types of histone methylation have been associated with cancer progression.Depending on the methylation site in histone proteins, its effects on transcription are different.Its knockdown by siRNA decreased the proliferation, migration, and invasion of gastric cancer cells, whereas its overexpression showed the opposite effects.

View Article: PubMed Central - PubMed

Affiliation: Departments of Anatomy, School of Medicine, Pusan National University, Busan, Republic of Korea.

ABSTRACT
Various types of histone methylation have been associated with cancer progression. Depending on the methylation site in histone proteins, its effects on transcription are different. DPY30 is a common member of SET1/MLL histone H3K4 methyltransferase complexes. However, its expression and roles in gastric cancer have been poorly characterized. To determine whether DPY30 has pathophysiological roles in gastric cancer, its expression and roles were examined. Immunohistochemistry and real time PCR showed up-regulation of DPY30 expression in some gastric cancer cell lines and patients' tissues. Its knockdown by siRNA decreased the proliferation, migration, and invasion of gastric cancer cells, whereas its overexpression showed the opposite effects. These results indicate that DPY30 has critical roles in the proliferation, migration, and invasion of gastric cancer cells, and suggest DPY30 might be a therapeutic target in gastric cancer.

No MeSH data available.


Related in: MedlinePlus