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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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The mRNA and protein levels of filamin C were significantly reduced in GC cell lines and GC tissues compared with the gastric cell line and corresponding normal gastric tissues(A) The filamin C mRNA in GC cell lines was measured using qPCR. (B) The mRNA of both isoforms a and b of filamin C were detected in GC cell lines. (C) The filamin C protein levels in GC cell lines were analyzed by Western blot. (D) The mRNA of filamin C was measured by qPCR in 36 pairs of human GC and the normal gastric tissues. (E) Filamin C expression in GC tissues was analyzed using a commercial TMA (Shanghai Outdo Biotech, China). (F) Statistical analysis of the expression of filamin C in 28 GC tissues and 13 normal gastric tissues included in a commercial TMA sample collection. (G) Statistical analysis of the expression of filamin C in 90 GC tissues and 90 normal gastric tissues included in another commercial TMA sample collection. (H) A Western blot analysis of filamin C expression in four pairs of GC samples and normal gastric tissues for which the mRNA levels were determined using qPCR as shown in Figure 2D.
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Figure 2: The mRNA and protein levels of filamin C were significantly reduced in GC cell lines and GC tissues compared with the gastric cell line and corresponding normal gastric tissues(A) The filamin C mRNA in GC cell lines was measured using qPCR. (B) The mRNA of both isoforms a and b of filamin C were detected in GC cell lines. (C) The filamin C protein levels in GC cell lines were analyzed by Western blot. (D) The mRNA of filamin C was measured by qPCR in 36 pairs of human GC and the normal gastric tissues. (E) Filamin C expression in GC tissues was analyzed using a commercial TMA (Shanghai Outdo Biotech, China). (F) Statistical analysis of the expression of filamin C in 28 GC tissues and 13 normal gastric tissues included in a commercial TMA sample collection. (G) Statistical analysis of the expression of filamin C in 90 GC tissues and 90 normal gastric tissues included in another commercial TMA sample collection. (H) A Western blot analysis of filamin C expression in four pairs of GC samples and normal gastric tissues for which the mRNA levels were determined using qPCR as shown in Figure 2D.

Mentions: To verify the results of proteomic analyses, we compared the expression of filamin C in GES-1 and 6 GC cell lines including the three used in the LC-MS analysis using quantitative reverse transcription (qPCR) and Western blot. Based on qPCR assay, the mRNA of filamin C was dramatically down-regulated in all six GC cell lines (Figure 2A). In addition, the results of qPCR with isoform-specific primers (Supplementary Table 1, Supplementary Fig. 1) demonstrated that both isoforms of filamin C were significantly down-regulated in all GC cell lines (Figure B). In addition, the protein level of filamin C in the 6 GC cell lines was significantly lower than that in GES-1 cells based on Western blot analysis, which was consistent with the qPCR results (Figure 2C).


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)

The mRNA and protein levels of filamin C were significantly reduced in GC cell lines and GC tissues compared with the gastric cell line and corresponding normal gastric tissues(A) The filamin C mRNA in GC cell lines was measured using qPCR. (B) The mRNA of both isoforms a and b of filamin C were detected in GC cell lines. (C) The filamin C protein levels in GC cell lines were analyzed by Western blot. (D) The mRNA of filamin C was measured by qPCR in 36 pairs of human GC and the normal gastric tissues. (E) Filamin C expression in GC tissues was analyzed using a commercial TMA (Shanghai Outdo Biotech, China). (F) Statistical analysis of the expression of filamin C in 28 GC tissues and 13 normal gastric tissues included in a commercial TMA sample collection. (G) Statistical analysis of the expression of filamin C in 90 GC tissues and 90 normal gastric tissues included in another commercial TMA sample collection. (H) A Western blot analysis of filamin C expression in four pairs of GC samples and normal gastric tissues for which the mRNA levels were determined using qPCR as shown in Figure 2D.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 2: The mRNA and protein levels of filamin C were significantly reduced in GC cell lines and GC tissues compared with the gastric cell line and corresponding normal gastric tissues(A) The filamin C mRNA in GC cell lines was measured using qPCR. (B) The mRNA of both isoforms a and b of filamin C were detected in GC cell lines. (C) The filamin C protein levels in GC cell lines were analyzed by Western blot. (D) The mRNA of filamin C was measured by qPCR in 36 pairs of human GC and the normal gastric tissues. (E) Filamin C expression in GC tissues was analyzed using a commercial TMA (Shanghai Outdo Biotech, China). (F) Statistical analysis of the expression of filamin C in 28 GC tissues and 13 normal gastric tissues included in a commercial TMA sample collection. (G) Statistical analysis of the expression of filamin C in 90 GC tissues and 90 normal gastric tissues included in another commercial TMA sample collection. (H) A Western blot analysis of filamin C expression in four pairs of GC samples and normal gastric tissues for which the mRNA levels were determined using qPCR as shown in Figure 2D.
Mentions: To verify the results of proteomic analyses, we compared the expression of filamin C in GES-1 and 6 GC cell lines including the three used in the LC-MS analysis using quantitative reverse transcription (qPCR) and Western blot. Based on qPCR assay, the mRNA of filamin C was dramatically down-regulated in all six GC cell lines (Figure 2A). In addition, the results of qPCR with isoform-specific primers (Supplementary Table 1, Supplementary Fig. 1) demonstrated that both isoforms of filamin C were significantly down-regulated in all GC cell lines (Figure B). In addition, the protein level of filamin C in the 6 GC cell lines was significantly lower than that in GES-1 cells based on Western blot analysis, which was consistent with the qPCR results (Figure 2C).

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