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TFPI-2 is a putative tumor suppressor gene frequently inactivated by promoter hypermethylation in nasopharyngeal carcinoma.

Wang S, Xiao X, Zhou X, Huang T, Du C, Yu N, Mo Y, Lin L, Zhang J, Ma N, Murata M, Huang G, Zhang Z - BMC Cancer (2010)

Bottom Line: TFPI-2 was aberrantly methylated in 66.7% (4/6) NPC cell lines and 88.6% (62/70) of NPC primary tumors, but not in normal nasopharyngeal epithelia.TFPI-2 expression could be restored in NPC cells after demethylation treatment.Ectopic expression of TFPI-2 in NPC cells induced apoptosis and inhibited cell proliferation, colony formation and cell migration.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Otolaryngology Head and Neck Surgery, First Affiliated Hospital of Guangxi Medical University, Nanning, PR China.

ABSTRACT

Background: Epigenetic silencing of tumor suppressor genes play important roles in NPC tumorgenesis. Tissue factor pathway inhibitor-2 (TFPI-2), is a protease inhibitor. Recently, TFPI-2 was suggested to be a tumor suppressor gene involved in tumorigenesis and metastasis in some cancers. In this study, we investigated whether TFPI-2 was inactivated epigenetically in nasopharyngeal carcinoma (NPC).

Methods: Transcriptional expression levels of TFPI-2 was evaluated by RT-PCR. Methylation status were investigated by methylation specific PCR and bisulfate genomic sequencing. The role of TFPI-2 as a tumor suppressor gene in NPC was addressed by re-introducing TFPI-2 expression into the NPC cell line CNE2.

Results: TFPI-2 mRNA transcription was inactivated in NPC cell lines. TFPI-2 was aberrantly methylated in 66.7% (4/6) NPC cell lines and 88.6% (62/70) of NPC primary tumors, but not in normal nasopharyngeal epithelia. TFPI-2 expression could be restored in NPC cells after demethylation treatment. Ectopic expression of TFPI-2 in NPC cells induced apoptosis and inhibited cell proliferation, colony formation and cell migration.

Conclusions: Epigenetic inactivation of TFPI-2 by promoter hypermethylation is a frequent and tumor specific event in NPC. TFPI-2 might be considering as a putative tumor suppressor gene in NPC.

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Related in: MedlinePlus

TFPI-2 inhibits NPC cell proliferation. A: RT-PCR validation of stable transfectance of CNE2-TFPI-2 or CNE2-Empty vector. B: Proliferation curves of CNE2 cells, stable transfectants of CNE2-TFPI-2 and CNE2-empty vector. Cells were counted every 24 h for 6 days.
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Figure 6: TFPI-2 inhibits NPC cell proliferation. A: RT-PCR validation of stable transfectance of CNE2-TFPI-2 or CNE2-Empty vector. B: Proliferation curves of CNE2 cells, stable transfectants of CNE2-TFPI-2 and CNE2-empty vector. Cells were counted every 24 h for 6 days.

Mentions: To assess whether TFPI-2 might possess properties as a tumor suppressor gene in NPC, we examined the effect of TFPI-2 on clonogenicity, cell proliferation, and cell mobility. The colony-formation efficiencies were evaluated by monolayer culture. The number of colonies formed by CNE2-TFPI-2 cells was less than that by CNE2-empty vector cells (p < 0.05) (Figure 7). This finding was further supported by cell proliferation assay. CNE2-TFPI-2 cells grew significantly slower than CNE2 parental cells and CNE2-empty vector cells (Figure 6). Wound healing assay was carried out to measure cell mobility by comparing the scratching healing efficiency of CNE2-TFPI-2 cells and CNE2 empty vector cells. The CNE2-TFPI-2 cells moved slower into the scratched areas than did CNE2-empty vector control cells, which suggests that ectopic expression of TFPI-2 inhibits the motility of NPC cells (Figure 8).


TFPI-2 is a putative tumor suppressor gene frequently inactivated by promoter hypermethylation in nasopharyngeal carcinoma.

Wang S, Xiao X, Zhou X, Huang T, Du C, Yu N, Mo Y, Lin L, Zhang J, Ma N, Murata M, Huang G, Zhang Z - BMC Cancer (2010)

TFPI-2 inhibits NPC cell proliferation. A: RT-PCR validation of stable transfectance of CNE2-TFPI-2 or CNE2-Empty vector. B: Proliferation curves of CNE2 cells, stable transfectants of CNE2-TFPI-2 and CNE2-empty vector. Cells were counted every 24 h for 6 days.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2992524&req=5

Figure 6: TFPI-2 inhibits NPC cell proliferation. A: RT-PCR validation of stable transfectance of CNE2-TFPI-2 or CNE2-Empty vector. B: Proliferation curves of CNE2 cells, stable transfectants of CNE2-TFPI-2 and CNE2-empty vector. Cells were counted every 24 h for 6 days.
Mentions: To assess whether TFPI-2 might possess properties as a tumor suppressor gene in NPC, we examined the effect of TFPI-2 on clonogenicity, cell proliferation, and cell mobility. The colony-formation efficiencies were evaluated by monolayer culture. The number of colonies formed by CNE2-TFPI-2 cells was less than that by CNE2-empty vector cells (p < 0.05) (Figure 7). This finding was further supported by cell proliferation assay. CNE2-TFPI-2 cells grew significantly slower than CNE2 parental cells and CNE2-empty vector cells (Figure 6). Wound healing assay was carried out to measure cell mobility by comparing the scratching healing efficiency of CNE2-TFPI-2 cells and CNE2 empty vector cells. The CNE2-TFPI-2 cells moved slower into the scratched areas than did CNE2-empty vector control cells, which suggests that ectopic expression of TFPI-2 inhibits the motility of NPC cells (Figure 8).

Bottom Line: TFPI-2 was aberrantly methylated in 66.7% (4/6) NPC cell lines and 88.6% (62/70) of NPC primary tumors, but not in normal nasopharyngeal epithelia.TFPI-2 expression could be restored in NPC cells after demethylation treatment.Ectopic expression of TFPI-2 in NPC cells induced apoptosis and inhibited cell proliferation, colony formation and cell migration.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Otolaryngology Head and Neck Surgery, First Affiliated Hospital of Guangxi Medical University, Nanning, PR China.

ABSTRACT

Background: Epigenetic silencing of tumor suppressor genes play important roles in NPC tumorgenesis. Tissue factor pathway inhibitor-2 (TFPI-2), is a protease inhibitor. Recently, TFPI-2 was suggested to be a tumor suppressor gene involved in tumorigenesis and metastasis in some cancers. In this study, we investigated whether TFPI-2 was inactivated epigenetically in nasopharyngeal carcinoma (NPC).

Methods: Transcriptional expression levels of TFPI-2 was evaluated by RT-PCR. Methylation status were investigated by methylation specific PCR and bisulfate genomic sequencing. The role of TFPI-2 as a tumor suppressor gene in NPC was addressed by re-introducing TFPI-2 expression into the NPC cell line CNE2.

Results: TFPI-2 mRNA transcription was inactivated in NPC cell lines. TFPI-2 was aberrantly methylated in 66.7% (4/6) NPC cell lines and 88.6% (62/70) of NPC primary tumors, but not in normal nasopharyngeal epithelia. TFPI-2 expression could be restored in NPC cells after demethylation treatment. Ectopic expression of TFPI-2 in NPC cells induced apoptosis and inhibited cell proliferation, colony formation and cell migration.

Conclusions: Epigenetic inactivation of TFPI-2 by promoter hypermethylation is a frequent and tumor specific event in NPC. TFPI-2 might be considering as a putative tumor suppressor gene in NPC.

Show MeSH
Related in: MedlinePlus