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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

Proteomic analyses of a normal gastric cell line (GES-1) and three GC cell lines (SGC-7901, MGC-803, and HGC-27)(A) The general procedures of proteomic analyses. (B) The total number of proteins identified in each cell line and the overlaps among the four cell lines. (C) The Venn diagrams showing the overlaps of the differentially expressed proteins among the three GC cell lines. The differentially expressed proteins in each GC cell line were determined with a fold change log2 ratio ≥ 1 (i.e. fold change ≥ 2) and a p value of the Student's t test < 0.05 or < 0.01. To calculate the fold changes, the average expressions in the GC cell sets were divided by the average expressions in the GES-1 set and the ratios were Log2 transformed. (D) The volcano plots depict the differentially expressed proteins in the three GC cell lines. The p values of t-test (−log10 transformed) were displayed as a function of the log2 transformed fold changes. The red dash lines indicated the threshold of p < 0.01 and fold change ≥ 2. The accession numbers of filamin C (Q14315) and UCHL1 (P09936) were highlighted.
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Figure 1: Proteomic analyses of a normal gastric cell line (GES-1) and three GC cell lines (SGC-7901, MGC-803, and HGC-27)(A) The general procedures of proteomic analyses. (B) The total number of proteins identified in each cell line and the overlaps among the four cell lines. (C) The Venn diagrams showing the overlaps of the differentially expressed proteins among the three GC cell lines. The differentially expressed proteins in each GC cell line were determined with a fold change log2 ratio ≥ 1 (i.e. fold change ≥ 2) and a p value of the Student's t test < 0.05 or < 0.01. To calculate the fold changes, the average expressions in the GC cell sets were divided by the average expressions in the GES-1 set and the ratios were Log2 transformed. (D) The volcano plots depict the differentially expressed proteins in the three GC cell lines. The p values of t-test (−log10 transformed) were displayed as a function of the log2 transformed fold changes. The red dash lines indicated the threshold of p < 0.01 and fold change ≥ 2. The accession numbers of filamin C (Q14315) and UCHL1 (P09936) were highlighted.

Mentions: GES-1 is an immortalized stomach mucosal cell line established by SV40 virus infection of 9 month human fetal gastric epithelial cells [23], whereas SGC-7901, MGC-803, and HGC-27 represent the moderate-, low- and non-differentiated gastric cancer cell lines, respectively. The proteomic profiles of the four cell lines were analyzed using label free LC-MS with LTQ Obitrap in triplicates (Figure 1A, Supplementary Methods and Supplementary Table 2). A total of 2,787 proteins including 36 decoy hits were identified from 27,067 distinct peptides and 347,681 tandem spectra. The false discovery rates at protein and spectrum level reported by Scaffold were 1.3% and 0.03%, respectively. The information of identified peptides and proteins were shown in Supplementary Tables 3 and 4, respectively. Among the 2,750 proteins, 1,395, 2,165, 2,271, and 1,478 proteins were identified in GES-1, SGC-7901, MGC-803, and HGC-27, respectively, and 1,065 proteins were shared by the four cell lines (Figure 1B).


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)

Proteomic analyses of a normal gastric cell line (GES-1) and three GC cell lines (SGC-7901, MGC-803, and HGC-27)(A) The general procedures of proteomic analyses. (B) The total number of proteins identified in each cell line and the overlaps among the four cell lines. (C) The Venn diagrams showing the overlaps of the differentially expressed proteins among the three GC cell lines. The differentially expressed proteins in each GC cell line were determined with a fold change log2 ratio ≥ 1 (i.e. fold change ≥ 2) and a p value of the Student's t test < 0.05 or < 0.01. To calculate the fold changes, the average expressions in the GC cell sets were divided by the average expressions in the GES-1 set and the ratios were Log2 transformed. (D) The volcano plots depict the differentially expressed proteins in the three GC cell lines. The p values of t-test (−log10 transformed) were displayed as a function of the log2 transformed fold changes. The red dash lines indicated the threshold of p < 0.01 and fold change ≥ 2. The accession numbers of filamin C (Q14315) and UCHL1 (P09936) were highlighted.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 1: Proteomic analyses of a normal gastric cell line (GES-1) and three GC cell lines (SGC-7901, MGC-803, and HGC-27)(A) The general procedures of proteomic analyses. (B) The total number of proteins identified in each cell line and the overlaps among the four cell lines. (C) The Venn diagrams showing the overlaps of the differentially expressed proteins among the three GC cell lines. The differentially expressed proteins in each GC cell line were determined with a fold change log2 ratio ≥ 1 (i.e. fold change ≥ 2) and a p value of the Student's t test < 0.05 or < 0.01. To calculate the fold changes, the average expressions in the GC cell sets were divided by the average expressions in the GES-1 set and the ratios were Log2 transformed. (D) The volcano plots depict the differentially expressed proteins in the three GC cell lines. The p values of t-test (−log10 transformed) were displayed as a function of the log2 transformed fold changes. The red dash lines indicated the threshold of p < 0.01 and fold change ≥ 2. The accession numbers of filamin C (Q14315) and UCHL1 (P09936) were highlighted.
Mentions: GES-1 is an immortalized stomach mucosal cell line established by SV40 virus infection of 9 month human fetal gastric epithelial cells [23], whereas SGC-7901, MGC-803, and HGC-27 represent the moderate-, low- and non-differentiated gastric cancer cell lines, respectively. The proteomic profiles of the four cell lines were analyzed using label free LC-MS with LTQ Obitrap in triplicates (Figure 1A, Supplementary Methods and Supplementary Table 2). A total of 2,787 proteins including 36 decoy hits were identified from 27,067 distinct peptides and 347,681 tandem spectra. The false discovery rates at protein and spectrum level reported by Scaffold were 1.3% and 0.03%, respectively. The information of identified peptides and proteins were shown in Supplementary Tables 3 and 4, respectively. Among the 2,750 proteins, 1,395, 2,165, 2,271, and 1,478 proteins were identified in GES-1, SGC-7901, MGC-803, and HGC-27, respectively, and 1,065 proteins were shared by the four cell lines (Figure 1B).

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