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Depletion of C3orf1/TIMMDC1 inhibits migration and proliferation in 95D lung carcinoma cells.

Wu H, Wang W, Xu H - Int J Mol Sci (2014)

Bottom Line: We demonstrated that C3orf1 depletion significantly suppressed 95D cell growth and migration.We confirmed C3orf1 localization in the inner mitochondrial membrane and showed that mitochondrial viability, membrane potential, and ATPase activity were remarkably reduced upon depletion of C3orf1.Concurrently, expression of the migration-promoting gene NUPR1 was markedly reduced, as confirmed by real-time PCR.

View Article: PubMed Central - PubMed

Affiliation: School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, China. huiling@ujs.edu.cn.

ABSTRACT
In our previous study, we identified an association of high expression of c3orf1, also known as TIMMDC1 (translocase of inner mitochondrial membrane domain-containing protein 1), with metastatic characteristics in lung carcinoma cells. To investigate the preliminary function and mechanism of this mitochondrial protein, we depleted C3orf1 expression by introducing siRNA into 95D lung carcinoma cells. We demonstrated that C3orf1 depletion significantly suppressed 95D cell growth and migration. We confirmed C3orf1 localization in the inner mitochondrial membrane and showed that mitochondrial viability, membrane potential, and ATPase activity were remarkably reduced upon depletion of C3orf1. Microarray data indicated that genes involved in regulation of cell death, migration, and cell-cycle arrest were significantly altered after C3orf1 depletion for 48 h. The expression of genes involved in focal adhesion, ECM-receptor interaction, and p53-signaling pathways were notably altered. Furthermore, cell-cycle arrest genes such as CCNG2 and PTEN as well as genes involved in cell migration inhibition, such as TIMP3 and COL3A1, were upregulated after C3orf1 depletion in 95D cells. Concurrently, expression of the migration-promoting gene NUPR1 was markedly reduced, as confirmed by real-time PCR. We conclude that C3orf1 is critical for mitochondrial function, migration, and proliferation in 95D lung carcinoma cells. Depletion of C3orf1 inhibited cell migration and cell proliferation in association with upregulation of genes involved in cell-cycle arrest and cell migration inhibition. These results suggest that C3orf1 (TIMMDC1) may be a viable treatment target for lung carcinoma, and that further study of the role of this protein in lung carcinoma pathogenesis is justified.

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Related in: MedlinePlus

Alteration in the gene expression profile of C3orf1-depleted cells. Graphic representation of real-time PCR results depicting the expression of selected genes; * p < 0.05.
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ijms-15-20555-f004: Alteration in the gene expression profile of C3orf1-depleted cells. Graphic representation of real-time PCR results depicting the expression of selected genes; * p < 0.05.

Mentions: To investigate the molecular signature of C3orf1 depletion in 95D cells, Affymetrix Genechip Prime View Human Gene Expression Array was used according to the method described in a previous study [16]. Cells treated with c3orf1 or control siRNA were screened for differentially expressed genes. We identified 439 upregulated probe sets corresponding to 360 unique genes and open reading frames and 98 downregulated probe sets corresponding to 61 genes and open reading frames (fold change > 2) in the c3orf1 siRNA treated cells compared to the control siRNA treated cells. Using gene ontology (GO) analysis (Table 1), these genes were identified as being involved in several biological processes related to tumor suppression, such as regulation of apoptosis, programmed cell death, cell-cycle arrest, and regulation of cell migration. With Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment, several of the altered pathways that were most likely to be involved in tumor invasion were identified, including focal adhesion and ECM-receptor interaction pathways (Table 2). Interestingly, the p53 signaling pathway, which is associated with cell proliferation, is involved. Based on these results, we chose transcripts of interest, such as TIMP3, NUPR1, COL3A1, CCNG2, and PTEN, for confirmation of array results by using real-time PCR. As shown in Figure 4 and Table 3, the cell-cycle arrest-related transcripts CCNG2 [17] and PTEN [18] were significantly upregulated in c3orf1 siRNA-treated cells compared to control cells (p < 0.05). Transcripts involved in cell migration inhibition, TIMP3 [19] and COL3A1 [20], were upregulated. The NUPR1 transcript [21], a migration promoting gene, was markedly downregulated upon C3orf1 depletion in 95D cells.


Depletion of C3orf1/TIMMDC1 inhibits migration and proliferation in 95D lung carcinoma cells.

Wu H, Wang W, Xu H - Int J Mol Sci (2014)

Alteration in the gene expression profile of C3orf1-depleted cells. Graphic representation of real-time PCR results depicting the expression of selected genes; * p < 0.05.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4264183&req=5

ijms-15-20555-f004: Alteration in the gene expression profile of C3orf1-depleted cells. Graphic representation of real-time PCR results depicting the expression of selected genes; * p < 0.05.
Mentions: To investigate the molecular signature of C3orf1 depletion in 95D cells, Affymetrix Genechip Prime View Human Gene Expression Array was used according to the method described in a previous study [16]. Cells treated with c3orf1 or control siRNA were screened for differentially expressed genes. We identified 439 upregulated probe sets corresponding to 360 unique genes and open reading frames and 98 downregulated probe sets corresponding to 61 genes and open reading frames (fold change > 2) in the c3orf1 siRNA treated cells compared to the control siRNA treated cells. Using gene ontology (GO) analysis (Table 1), these genes were identified as being involved in several biological processes related to tumor suppression, such as regulation of apoptosis, programmed cell death, cell-cycle arrest, and regulation of cell migration. With Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment, several of the altered pathways that were most likely to be involved in tumor invasion were identified, including focal adhesion and ECM-receptor interaction pathways (Table 2). Interestingly, the p53 signaling pathway, which is associated with cell proliferation, is involved. Based on these results, we chose transcripts of interest, such as TIMP3, NUPR1, COL3A1, CCNG2, and PTEN, for confirmation of array results by using real-time PCR. As shown in Figure 4 and Table 3, the cell-cycle arrest-related transcripts CCNG2 [17] and PTEN [18] were significantly upregulated in c3orf1 siRNA-treated cells compared to control cells (p < 0.05). Transcripts involved in cell migration inhibition, TIMP3 [19] and COL3A1 [20], were upregulated. The NUPR1 transcript [21], a migration promoting gene, was markedly downregulated upon C3orf1 depletion in 95D cells.

Bottom Line: We demonstrated that C3orf1 depletion significantly suppressed 95D cell growth and migration.We confirmed C3orf1 localization in the inner mitochondrial membrane and showed that mitochondrial viability, membrane potential, and ATPase activity were remarkably reduced upon depletion of C3orf1.Concurrently, expression of the migration-promoting gene NUPR1 was markedly reduced, as confirmed by real-time PCR.

View Article: PubMed Central - PubMed

Affiliation: School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, China. huiling@ujs.edu.cn.

ABSTRACT
In our previous study, we identified an association of high expression of c3orf1, also known as TIMMDC1 (translocase of inner mitochondrial membrane domain-containing protein 1), with metastatic characteristics in lung carcinoma cells. To investigate the preliminary function and mechanism of this mitochondrial protein, we depleted C3orf1 expression by introducing siRNA into 95D lung carcinoma cells. We demonstrated that C3orf1 depletion significantly suppressed 95D cell growth and migration. We confirmed C3orf1 localization in the inner mitochondrial membrane and showed that mitochondrial viability, membrane potential, and ATPase activity were remarkably reduced upon depletion of C3orf1. Microarray data indicated that genes involved in regulation of cell death, migration, and cell-cycle arrest were significantly altered after C3orf1 depletion for 48 h. The expression of genes involved in focal adhesion, ECM-receptor interaction, and p53-signaling pathways were notably altered. Furthermore, cell-cycle arrest genes such as CCNG2 and PTEN as well as genes involved in cell migration inhibition, such as TIMP3 and COL3A1, were upregulated after C3orf1 depletion in 95D cells. Concurrently, expression of the migration-promoting gene NUPR1 was markedly reduced, as confirmed by real-time PCR. We conclude that C3orf1 is critical for mitochondrial function, migration, and proliferation in 95D lung carcinoma cells. Depletion of C3orf1 inhibited cell migration and cell proliferation in association with upregulation of genes involved in cell-cycle arrest and cell migration inhibition. These results suggest that C3orf1 (TIMMDC1) may be a viable treatment target for lung carcinoma, and that further study of the role of this protein in lung carcinoma pathogenesis is justified.

Show MeSH
Related in: MedlinePlus