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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 proliferation of GC cells(A) Ectopic expression of filamin C in SGC-7901 and HGC-27 cells cells inhibited the proliferation of cancer cells based on BrdU assay after two days of culturing. The filamin C overexpression was confirmed using Western blot (left). (B) Knockdown of filamin C by two shRNAs (sh1 and sh4) improved the proliferation of cancer cell line DU145. Filamin C knockdown was confirmed using Western blot (left). (C) Clone formation assay of the proliferation of DU145 cells in which endogenous filamin C was silenced by shRNA. The statistical analysis results of the clone numbers of DU145 cells were shown on the right. Statistical significances were calculated using Student's t tests and a p value < 0.05 was considered as statistically significant. FLNC, filamin C.
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Figure 4: Filamin C inhibited the proliferation of GC cells(A) Ectopic expression of filamin C in SGC-7901 and HGC-27 cells cells inhibited the proliferation of cancer cells based on BrdU assay after two days of culturing. The filamin C overexpression was confirmed using Western blot (left). (B) Knockdown of filamin C by two shRNAs (sh1 and sh4) improved the proliferation of cancer cell line DU145. Filamin C knockdown was confirmed using Western blot (left). (C) Clone formation assay of the proliferation of DU145 cells in which endogenous filamin C was silenced by shRNA. The statistical analysis results of the clone numbers of DU145 cells were shown on the right. Statistical significances were calculated using Student's t tests and a p value < 0.05 was considered as statistically significant. FLNC, filamin C.

Mentions: In addition, the expression of filamin C was significantly reduced in 36 GC tissues compared with their corresponding normal gastric tissues based on qPCR assay (p = 0.002, n = 36) (Figure 4D). The expression of filamin C was also evaluated in human GC tissues from two commercial tissue microarrays (TMAs) containing different sets of GC cases. Consistently, filamin C expression was obviously reduced in these GC tissues (p = 1.1E-6) compared with normal gastric tissues (Figure 2E and F). In the analysis of another TMA containing 90 pairs GC and normal counterpart tissues, filamin C was significantly downregulated in GCs (p = 1.7–18, paired Student's t test) (Figure 2G). The sample information and scoring results for the staining were deposited in Supplementary Table 5. We also analyzed the protein expression of filamin C in GC tissues using Western blot. Filamin C was low in all four GC tissues comparing with the normal counterparts (Figure 2H).


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 proliferation of GC cells(A) Ectopic expression of filamin C in SGC-7901 and HGC-27 cells cells inhibited the proliferation of cancer cells based on BrdU assay after two days of culturing. The filamin C overexpression was confirmed using Western blot (left). (B) Knockdown of filamin C by two shRNAs (sh1 and sh4) improved the proliferation of cancer cell line DU145. Filamin C knockdown was confirmed using Western blot (left). (C) Clone formation assay of the proliferation of DU145 cells in which endogenous filamin C was silenced by shRNA. The statistical analysis results of the clone numbers of DU145 cells were shown on the right. Statistical significances were calculated using Student's t tests and a p value < 0.05 was considered as statistically significant. FLNC, filamin C.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 4: Filamin C inhibited the proliferation of GC cells(A) Ectopic expression of filamin C in SGC-7901 and HGC-27 cells cells inhibited the proliferation of cancer cells based on BrdU assay after two days of culturing. The filamin C overexpression was confirmed using Western blot (left). (B) Knockdown of filamin C by two shRNAs (sh1 and sh4) improved the proliferation of cancer cell line DU145. Filamin C knockdown was confirmed using Western blot (left). (C) Clone formation assay of the proliferation of DU145 cells in which endogenous filamin C was silenced by shRNA. The statistical analysis results of the clone numbers of DU145 cells were shown on the right. Statistical significances were calculated using Student's t tests and a p value < 0.05 was considered as statistically significant. FLNC, filamin C.
Mentions: In addition, the expression of filamin C was significantly reduced in 36 GC tissues compared with their corresponding normal gastric tissues based on qPCR assay (p = 0.002, n = 36) (Figure 4D). The expression of filamin C was also evaluated in human GC tissues from two commercial tissue microarrays (TMAs) containing different sets of GC cases. Consistently, filamin C expression was obviously reduced in these GC tissues (p = 1.1E-6) compared with normal gastric tissues (Figure 2E and F). In the analysis of another TMA containing 90 pairs GC and normal counterpart tissues, filamin C was significantly downregulated in GCs (p = 1.7–18, paired Student's t test) (Figure 2G). The sample information and scoring results for the staining were deposited in Supplementary Table 5. We also analyzed the protein expression of filamin C in GC tissues using Western blot. Filamin C was low in all four GC tissues comparing with the normal counterparts (Figure 2H).

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