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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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Related in: MedlinePlus

Filamin C inhibited the metastasis of cancer cells through downregulating MMP2(A) DU145 cells with filamin C silencing were fluorescence-labeled and analyzed in a zebrafish cancer metastasis model. The selected pictures were shown from 0 to 3 days post injection. The areas indicated with arrows were enlarged for visualizing disseminated cancer cells. (B)Filamin C was silenced in DU145 cells and the proenzyme (Pro) and activated form of MMP2 were analyzed. (C) Migration of DU145 cells with or without filamin C silencing were analyzed. The third group with filamin C silencing was further treated with the MMP2 inhibitor ARP100. The statistical analysis results were shown in Figure 6D and the Western blot results were shown in Figure 6E. FLNC, filamin C.
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Figure 6: Filamin C inhibited the metastasis of cancer cells through downregulating MMP2(A) DU145 cells with filamin C silencing were fluorescence-labeled and analyzed in a zebrafish cancer metastasis model. The selected pictures were shown from 0 to 3 days post injection. The areas indicated with arrows were enlarged for visualizing disseminated cancer cells. (B)Filamin C was silenced in DU145 cells and the proenzyme (Pro) and activated form of MMP2 were analyzed. (C) Migration of DU145 cells with or without filamin C silencing were analyzed. The third group with filamin C silencing was further treated with the MMP2 inhibitor ARP100. The statistical analysis results were shown in Figure 6D and the Western blot results were shown in Figure 6E. FLNC, filamin C.

Mentions: We then used a zebrafish cancer metastasis model to investigate the role of filamin C in metastasis of cancer cells in vivo. Fluorescence dye-labeled DU145 cells were inoculated into the yolk of 48 hpf zebrafish and cell dissemination was measured in the following 3 days. As shown in Figure 6A, DU145 cells with stable filamin C silencing disseminated in the fish york after 24 h post injection and continued to spread to the head and tail of fish compared with the fish inoculated with DU145 cells without filamin C silencing. Collectively, filamin C-sh1 group had 36 (73.5%) metastasis-positive and 13 (26.5%) metastasis-negative fish, while pLKO.1-shluciferase group had less metastasis-positive (n = 25, 45.5%) but more metastasis-negative (n = 30, 54.4%) fish (Table 2). Significant difference was identified between the two groups based on Chi-square test (p = 0.004).


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 metastasis of cancer cells through downregulating MMP2(A) DU145 cells with filamin C silencing were fluorescence-labeled and analyzed in a zebrafish cancer metastasis model. The selected pictures were shown from 0 to 3 days post injection. The areas indicated with arrows were enlarged for visualizing disseminated cancer cells. (B)Filamin C was silenced in DU145 cells and the proenzyme (Pro) and activated form of MMP2 were analyzed. (C) Migration of DU145 cells with or without filamin C silencing were analyzed. The third group with filamin C silencing was further treated with the MMP2 inhibitor ARP100. The statistical analysis results were shown in Figure 6D and the Western blot results were shown in Figure 6E. FLNC, filamin C.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4359225&req=5

Figure 6: Filamin C inhibited the metastasis of cancer cells through downregulating MMP2(A) DU145 cells with filamin C silencing were fluorescence-labeled and analyzed in a zebrafish cancer metastasis model. The selected pictures were shown from 0 to 3 days post injection. The areas indicated with arrows were enlarged for visualizing disseminated cancer cells. (B)Filamin C was silenced in DU145 cells and the proenzyme (Pro) and activated form of MMP2 were analyzed. (C) Migration of DU145 cells with or without filamin C silencing were analyzed. The third group with filamin C silencing was further treated with the MMP2 inhibitor ARP100. The statistical analysis results were shown in Figure 6D and the Western blot results were shown in Figure 6E. FLNC, filamin C.
Mentions: We then used a zebrafish cancer metastasis model to investigate the role of filamin C in metastasis of cancer cells in vivo. Fluorescence dye-labeled DU145 cells were inoculated into the yolk of 48 hpf zebrafish and cell dissemination was measured in the following 3 days. As shown in Figure 6A, DU145 cells with stable filamin C silencing disseminated in the fish york after 24 h post injection and continued to spread to the head and tail of fish compared with the fish inoculated with DU145 cells without filamin C silencing. Collectively, filamin C-sh1 group had 36 (73.5%) metastasis-positive and 13 (26.5%) metastasis-negative fish, while pLKO.1-shluciferase group had less metastasis-positive (n = 25, 45.5%) but more metastasis-negative (n = 30, 54.4%) fish (Table 2). Significant difference was identified between the two groups based on Chi-square test (p = 0.004).

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