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Overexpression of stathmin1 in the diffuse type of gastric cancer and its roles in proliferation and migration of gastric cancer cells.

Jeon TY, Han ME, Lee YW, Lee YS, Kim GH, Song GA, Hur GY, Kim JY, Kim HJ, Yoon S, Baek SY, Kim BS, Kim JB, Oh SO - Br. J. Cancer (2010)

Bottom Line: When the expression of stathmin1 was knocked down using siRNA, the proliferation, migration and invasion of poorly differentiated gastric cancer cells in vitro were significantly inhibited.Moreover, stathmin1 siRNA transfection significantly slowed the growth of xenografts in nude mice.These results suggest that stathmin1 can be a good prognostic factor for recurrence-free survival rate and is a therapeutic target in diffuse-type gastric cancer.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, Pusan National University, Beomeo-Ri, Mulgeum-Eup, Yangsan, South Korea.

ABSTRACT

Background: Stathmin1 is a microtubule-regulating protein that has an important role in the assembly and disassembly of the mitotic spindle. The roles of stathmin1 in carcinogenesis of various cancers, including prostate and breast cancer, have been explored. However, its expression and roles in gastric cancer have not yet been described.

Methods: Stathmin1 expression in paraffin-embedded tissue sections from 226 patients was analysed by immunohistochemistry. Roles of stathmin1 were studied using a specific small interfering RNA (siRNA).

Results: The expression of stathmin1 was positively correlated with lymph node metastasis, TNM stages and vascular invasion, and negatively with recurrence-free survival, in the diffuse type of gastric cancer. The median recurrence-free survival in patients with a negative and positive expression of stathmin1 was 17.0 and 7.0 months, respectively (P=0.009). When the expression of stathmin1 was knocked down using siRNA, the proliferation, migration and invasion of poorly differentiated gastric cancer cells in vitro were significantly inhibited. Moreover, stathmin1 siRNA transfection significantly slowed the growth of xenografts in nude mice.

Conclusion: These results suggest that stathmin1 can be a good prognostic factor for recurrence-free survival rate and is a therapeutic target in diffuse-type gastric cancer.

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Effect of stathmin1 silencing on the migration and invasion of gastric cancer cells. (A) Cell migration was evaluated in the Boyden migration assay two days after SNU638 cells were transfected with scrambled (SCR) small interfering RNA (siRNA) or stathmin1 siRNA. (B) Cell invasion was evaluated in the Matrigel invasion assay as described in the ‘Materials and Methods' section. Data are expressed as percentage change (means±s.d.) compared with controls and represent four independent experiments. (*P<0.01 vs SCR siRNA, one-way analysis of variance (ANOVA) followed by Tukey's multiple comparison). Representative microscopic images were presented in the upper panel of each assay graph. ANOVA, one-way analysis of variance; SCR, scrambled; siRNA, small interfering RNA.
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fig6: Effect of stathmin1 silencing on the migration and invasion of gastric cancer cells. (A) Cell migration was evaluated in the Boyden migration assay two days after SNU638 cells were transfected with scrambled (SCR) small interfering RNA (siRNA) or stathmin1 siRNA. (B) Cell invasion was evaluated in the Matrigel invasion assay as described in the ‘Materials and Methods' section. Data are expressed as percentage change (means±s.d.) compared with controls and represent four independent experiments. (*P<0.01 vs SCR siRNA, one-way analysis of variance (ANOVA) followed by Tukey's multiple comparison). Representative microscopic images were presented in the upper panel of each assay graph. ANOVA, one-way analysis of variance; SCR, scrambled; siRNA, small interfering RNA.

Mentions: To examine the possible roles of stathmin1 in gastric cancer cells, we knocked down the expression of stathmin1 using siRNA. Before siRNA experiments, we confirmed stathmin1 expression in gastric cancer cell lines by western blotting (data not shown). To confirm the efficacy of stathmin1 siRNA, we performed real-time PCR and western blotting (Figure 4). When SNU638 and SNU16 cells were transfected with stathmin1 siRNA, the expression of stathmin1 was almost completely abolished at the protein level (Figure 4A). Moreover, the messenger RNA level was also significantly reduced by siRNA (Figure 4B). To examine the role of stathmin1 in proliferation, we conducted WST assays after the transfection of stathmin1 siRNA. Stathmin1 siRNA significantly reduced the proliferation of SNU638 and SNU16 cells compared with SCR siRNA at 100 nM (Figure 5A and B). We next examined the role of stathmin1 in cancer cell migration. We observed a significant difference between cells transfected with SCR or stathmin1 siRNA in the migration assay (Figure 6A). Moreover, cancer cell invasion was also significantly reduced by stathmin1 siRNA in the Matrigel invasion assay (Figure 6B).


Overexpression of stathmin1 in the diffuse type of gastric cancer and its roles in proliferation and migration of gastric cancer cells.

Jeon TY, Han ME, Lee YW, Lee YS, Kim GH, Song GA, Hur GY, Kim JY, Kim HJ, Yoon S, Baek SY, Kim BS, Kim JB, Oh SO - Br. J. Cancer (2010)

Effect of stathmin1 silencing on the migration and invasion of gastric cancer cells. (A) Cell migration was evaluated in the Boyden migration assay two days after SNU638 cells were transfected with scrambled (SCR) small interfering RNA (siRNA) or stathmin1 siRNA. (B) Cell invasion was evaluated in the Matrigel invasion assay as described in the ‘Materials and Methods' section. Data are expressed as percentage change (means±s.d.) compared with controls and represent four independent experiments. (*P<0.01 vs SCR siRNA, one-way analysis of variance (ANOVA) followed by Tukey's multiple comparison). Representative microscopic images were presented in the upper panel of each assay graph. ANOVA, one-way analysis of variance; SCR, scrambled; siRNA, small interfering RNA.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2837578&req=5

fig6: Effect of stathmin1 silencing on the migration and invasion of gastric cancer cells. (A) Cell migration was evaluated in the Boyden migration assay two days after SNU638 cells were transfected with scrambled (SCR) small interfering RNA (siRNA) or stathmin1 siRNA. (B) Cell invasion was evaluated in the Matrigel invasion assay as described in the ‘Materials and Methods' section. Data are expressed as percentage change (means±s.d.) compared with controls and represent four independent experiments. (*P<0.01 vs SCR siRNA, one-way analysis of variance (ANOVA) followed by Tukey's multiple comparison). Representative microscopic images were presented in the upper panel of each assay graph. ANOVA, one-way analysis of variance; SCR, scrambled; siRNA, small interfering RNA.
Mentions: To examine the possible roles of stathmin1 in gastric cancer cells, we knocked down the expression of stathmin1 using siRNA. Before siRNA experiments, we confirmed stathmin1 expression in gastric cancer cell lines by western blotting (data not shown). To confirm the efficacy of stathmin1 siRNA, we performed real-time PCR and western blotting (Figure 4). When SNU638 and SNU16 cells were transfected with stathmin1 siRNA, the expression of stathmin1 was almost completely abolished at the protein level (Figure 4A). Moreover, the messenger RNA level was also significantly reduced by siRNA (Figure 4B). To examine the role of stathmin1 in proliferation, we conducted WST assays after the transfection of stathmin1 siRNA. Stathmin1 siRNA significantly reduced the proliferation of SNU638 and SNU16 cells compared with SCR siRNA at 100 nM (Figure 5A and B). We next examined the role of stathmin1 in cancer cell migration. We observed a significant difference between cells transfected with SCR or stathmin1 siRNA in the migration assay (Figure 6A). Moreover, cancer cell invasion was also significantly reduced by stathmin1 siRNA in the Matrigel invasion assay (Figure 6B).

Bottom Line: When the expression of stathmin1 was knocked down using siRNA, the proliferation, migration and invasion of poorly differentiated gastric cancer cells in vitro were significantly inhibited.Moreover, stathmin1 siRNA transfection significantly slowed the growth of xenografts in nude mice.These results suggest that stathmin1 can be a good prognostic factor for recurrence-free survival rate and is a therapeutic target in diffuse-type gastric cancer.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, Pusan National University, Beomeo-Ri, Mulgeum-Eup, Yangsan, South Korea.

ABSTRACT

Background: Stathmin1 is a microtubule-regulating protein that has an important role in the assembly and disassembly of the mitotic spindle. The roles of stathmin1 in carcinogenesis of various cancers, including prostate and breast cancer, have been explored. However, its expression and roles in gastric cancer have not yet been described.

Methods: Stathmin1 expression in paraffin-embedded tissue sections from 226 patients was analysed by immunohistochemistry. Roles of stathmin1 were studied using a specific small interfering RNA (siRNA).

Results: The expression of stathmin1 was positively correlated with lymph node metastasis, TNM stages and vascular invasion, and negatively with recurrence-free survival, in the diffuse type of gastric cancer. The median recurrence-free survival in patients with a negative and positive expression of stathmin1 was 17.0 and 7.0 months, respectively (P=0.009). When the expression of stathmin1 was knocked down using siRNA, the proliferation, migration and invasion of poorly differentiated gastric cancer cells in vitro were significantly inhibited. Moreover, stathmin1 siRNA transfection significantly slowed the growth of xenografts in nude mice.

Conclusion: These results suggest that stathmin1 can be a good prognostic factor for recurrence-free survival rate and is a therapeutic target in diffuse-type gastric cancer.

Show MeSH
Related in: MedlinePlus