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Filamin C, a dysregulated protein in cancer revealed by label-free quantitative proteomic analyses of human gastric cancer cells.

Qiao J, Cui SJ, Xu LL, Chen SJ, Yao J, Jiang YH, Peng G, Fang CY, Yang PY, Liu F - Oncotarget (2015)

Bottom Line: Nine proteins were significantly dysregulated in all three GC cell lines, including filamin C, a muscle-specific filamin and a large actin-cross-linking protein.Silencing of filamin C increased the expression of matrix metallopeptidase 2 and improved the metastasis of prostate cancer in a zebrafish model.High filamin C associated with better prognosis of prostate cancer, leukemia and breast cancer patients.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Systems Biology of School of Basic Medical Sciences and Institutes of Biomedical Sciences, Fudan University, Shanghai 200032, China.

ABSTRACT
Gastric cancer (GC) is the fourth and fifth most common cancer in men and women, respectively. We identified 2,750 proteins at false discovery rates of 1.3% (protein) and 0.03% (spectrum) by comparing the proteomic profiles of three GC and a normal gastric cell lines. Nine proteins were significantly dysregulated in all three GC cell lines, including filamin C, a muscle-specific filamin and a large actin-cross-linking protein. Downregulation of filamin C in GC cell lines and tissues were verified using quantitative PCR and immunohistochemistry. Data-mining using public microarray datasets shown that filamin C was significantly reduced in many human primary and metastasis cancers. Transient expression or silencing of filamin C affected the proliferation and colony formation of cancer cells. Silencing of endogenous filamin C enhanced cancer cell migration and invasion, whereas ectopic expression of filamin C had opposing effects. Silencing of filamin C increased the expression of matrix metallopeptidase 2 and improved the metastasis of prostate cancer in a zebrafish model. High filamin C associated with better prognosis of prostate cancer, leukemia and breast cancer patients. These findings establish a functional role of filamin C in human cancers and these data will be valuable for further study of its mechanisms.

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Filamin C inhibited the migration and invasion of cancer cells(A)Filamin C silencing was performed in GES-1 cells, which express relatively high level of endogenous filamin C. Cell migration ability was analyzed in Transwell plates and representative images of the migration cells were shown. Western blots in the middle panel showed the results of filamin C silencing. The cells that migrated to the bottom surface of the membrane were stained by crystal violet, which was solubilized with acetic acid and absorbance was measured under 570 nm (right panel). siNC, negative control of silencing. (B)Filamin C gene was ectopically expressed in GC cell lines and cell migration ability was analyzed in a Transwell plate. Overexpression of filamin C was shown in the middle panel. Cell migration was measured using the absorbance method. Statistical analysis was performed using Student's t test and p < 0.05 was considered as statistically significant. 3.1, pcDNA3.1 vector; 3.1-filamin C, expression vector pcDNA3.1-filamin C. (C) Western blot analysis indicated that filamin C expression in the prostate cancer cell line DU145 was similar to that in GC cell line GES-1. Filamin C silencing in the prostate cancer cell lines DU145 and PC-3 improved cell migration (D) and invasion (E). Consistent results were obtained from triplicate experiments. FLNC, filamin C.
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Figure 5: Filamin C inhibited the migration and invasion of cancer cells(A)Filamin C silencing was performed in GES-1 cells, which express relatively high level of endogenous filamin C. Cell migration ability was analyzed in Transwell plates and representative images of the migration cells were shown. Western blots in the middle panel showed the results of filamin C silencing. The cells that migrated to the bottom surface of the membrane were stained by crystal violet, which was solubilized with acetic acid and absorbance was measured under 570 nm (right panel). siNC, negative control of silencing. (B)Filamin C gene was ectopically expressed in GC cell lines and cell migration ability was analyzed in a Transwell plate. Overexpression of filamin C was shown in the middle panel. Cell migration was measured using the absorbance method. Statistical analysis was performed using Student's t test and p < 0.05 was considered as statistically significant. 3.1, pcDNA3.1 vector; 3.1-filamin C, expression vector pcDNA3.1-filamin C. (C) Western blot analysis indicated that filamin C expression in the prostate cancer cell line DU145 was similar to that in GC cell line GES-1. Filamin C silencing in the prostate cancer cell lines DU145 and PC-3 improved cell migration (D) and invasion (E). Consistent results were obtained from triplicate experiments. FLNC, filamin C.

Mentions: Silencing endogenous filamin C by siRNA in GES-1 cells significantly improved the the migration ability of GES-1 cells (p = 0.03) (Figure 5A). On the contrary, re-expression of filamin C significantly inhibited the migration of GC cell lines SGC-7901 (p = 0.01), AGS (p = 0.011), and MGC-803 (p = 0.021) (Figure 5B). As mentioned above, the prostate cancer cell line DU145 has a relatively high level of filamin C expression, which is comparable to that in the normal gastric cells GES-1 (Figure 5C). Silencing of endogenous filamin C in DU145 cells significantly improved the migration of DU145 cells (p = 0.0007) (Figure 5D). Similarly, partial silencing of filamin C considerably increased the migration of prostate cancer cell line PC-3 (p = 0.021) (Figure 5D). In addition, Matrigel invasion assays demonstrated that filamin C knockdown enhanced the invasion of DU145 (p = 0.003) and PC-3 cells (p = 0.022) (Figure 5E).


Filamin C, a dysregulated protein in cancer revealed by label-free quantitative proteomic analyses of human gastric cancer cells.

Qiao J, Cui SJ, Xu LL, Chen SJ, Yao J, Jiang YH, Peng G, Fang CY, Yang PY, Liu F - Oncotarget (2015)

Filamin C inhibited the migration and invasion of cancer cells(A)Filamin C silencing was performed in GES-1 cells, which express relatively high level of endogenous filamin C. Cell migration ability was analyzed in Transwell plates and representative images of the migration cells were shown. Western blots in the middle panel showed the results of filamin C silencing. The cells that migrated to the bottom surface of the membrane were stained by crystal violet, which was solubilized with acetic acid and absorbance was measured under 570 nm (right panel). siNC, negative control of silencing. (B)Filamin C gene was ectopically expressed in GC cell lines and cell migration ability was analyzed in a Transwell plate. Overexpression of filamin C was shown in the middle panel. Cell migration was measured using the absorbance method. Statistical analysis was performed using Student's t test and p < 0.05 was considered as statistically significant. 3.1, pcDNA3.1 vector; 3.1-filamin C, expression vector pcDNA3.1-filamin C. (C) Western blot analysis indicated that filamin C expression in the prostate cancer cell line DU145 was similar to that in GC cell line GES-1. Filamin C silencing in the prostate cancer cell lines DU145 and PC-3 improved cell migration (D) and invasion (E). Consistent results were obtained from triplicate experiments. FLNC, filamin C.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Figure 5: Filamin C inhibited the migration and invasion of cancer cells(A)Filamin C silencing was performed in GES-1 cells, which express relatively high level of endogenous filamin C. Cell migration ability was analyzed in Transwell plates and representative images of the migration cells were shown. Western blots in the middle panel showed the results of filamin C silencing. The cells that migrated to the bottom surface of the membrane were stained by crystal violet, which was solubilized with acetic acid and absorbance was measured under 570 nm (right panel). siNC, negative control of silencing. (B)Filamin C gene was ectopically expressed in GC cell lines and cell migration ability was analyzed in a Transwell plate. Overexpression of filamin C was shown in the middle panel. Cell migration was measured using the absorbance method. Statistical analysis was performed using Student's t test and p < 0.05 was considered as statistically significant. 3.1, pcDNA3.1 vector; 3.1-filamin C, expression vector pcDNA3.1-filamin C. (C) Western blot analysis indicated that filamin C expression in the prostate cancer cell line DU145 was similar to that in GC cell line GES-1. Filamin C silencing in the prostate cancer cell lines DU145 and PC-3 improved cell migration (D) and invasion (E). Consistent results were obtained from triplicate experiments. FLNC, filamin C.
Mentions: Silencing endogenous filamin C by siRNA in GES-1 cells significantly improved the the migration ability of GES-1 cells (p = 0.03) (Figure 5A). On the contrary, re-expression of filamin C significantly inhibited the migration of GC cell lines SGC-7901 (p = 0.01), AGS (p = 0.011), and MGC-803 (p = 0.021) (Figure 5B). As mentioned above, the prostate cancer cell line DU145 has a relatively high level of filamin C expression, which is comparable to that in the normal gastric cells GES-1 (Figure 5C). Silencing of endogenous filamin C in DU145 cells significantly improved the migration of DU145 cells (p = 0.0007) (Figure 5D). Similarly, partial silencing of filamin C considerably increased the migration of prostate cancer cell line PC-3 (p = 0.021) (Figure 5D). In addition, Matrigel invasion assays demonstrated that filamin C knockdown enhanced the invasion of DU145 (p = 0.003) and PC-3 cells (p = 0.022) (Figure 5E).

Bottom Line: Nine proteins were significantly dysregulated in all three GC cell lines, including filamin C, a muscle-specific filamin and a large actin-cross-linking protein.Silencing of filamin C increased the expression of matrix metallopeptidase 2 and improved the metastasis of prostate cancer in a zebrafish model.High filamin C associated with better prognosis of prostate cancer, leukemia and breast cancer patients.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Systems Biology of School of Basic Medical Sciences and Institutes of Biomedical Sciences, Fudan University, Shanghai 200032, China.

ABSTRACT
Gastric cancer (GC) is the fourth and fifth most common cancer in men and women, respectively. We identified 2,750 proteins at false discovery rates of 1.3% (protein) and 0.03% (spectrum) by comparing the proteomic profiles of three GC and a normal gastric cell lines. Nine proteins were significantly dysregulated in all three GC cell lines, including filamin C, a muscle-specific filamin and a large actin-cross-linking protein. Downregulation of filamin C in GC cell lines and tissues were verified using quantitative PCR and immunohistochemistry. Data-mining using public microarray datasets shown that filamin C was significantly reduced in many human primary and metastasis cancers. Transient expression or silencing of filamin C affected the proliferation and colony formation of cancer cells. Silencing of endogenous filamin C enhanced cancer cell migration and invasion, whereas ectopic expression of filamin C had opposing effects. Silencing of filamin C increased the expression of matrix metallopeptidase 2 and improved the metastasis of prostate cancer in a zebrafish model. High filamin C associated with better prognosis of prostate cancer, leukemia and breast cancer patients. These findings establish a functional role of filamin C in human cancers and these data will be valuable for further study of its mechanisms.

Show MeSH
Related in: MedlinePlus