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Capn4 is a marker of poor clinical outcomes and promotes nasopharyngeal carcinoma metastasis via nuclear factor-κB-induced matrix metalloproteinase 2 expression.

Zheng PC, Chen X, Zhu HW, Zheng W, Mao LH, Lin C, Liu JN, Zheng M - Cancer Sci. (2014)

Bottom Line: The patients with NPC displaying higher Capn4 had a significantly shorter overall survival (P = 0.002) and progression-free survival (P = 0.003).These events resulted from Capn4 downregulation were associated with reduced expression of matrix metalloproteinase 2 (MMP2), Snail, and Vimentin.Together, these findings argue a novel function of Capn4 in invasion and metastasis of NPC, and thereby suggest that Capn4 may represent an independent prognostic factor and a potential therapeutic target in NPC.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy, School of Basic Medical Sciences, Fujian Medical University, Fuzhou, China.

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Knockdown of Capn4 reduces nasopharyngeal carcinoma (NPC)cell migration and invasion in vitro and in vivo. (a, b) 5-8F cells and CNE2 cells were stably transfected with constructs encoding siRNA specifically targeting Capn4 or scrambled sequence as siRNA control, after which semi-quantitative RT-PCR and Western blot analyses were performed to monitor mRNA and protein levels of Capn4 in Capn4 siRNA and control siRNA cells, as well as parental 5-8F cells and CNE2 cells. β-actin was probed as internal control. (c) Transwell assays were performed to measure in vitro migration and invasion of Capn4 siRNA and control siRNA cells. (d) To evaluate in vivo metastasis of Capn4 siRNA and control siRNA cells (n = 10 per group), 1 × 106 cells were resuspended in 0.1 mL of PBS and injected via the lateral tail vein. After 10 weeks, mice were euthanized, and metastatic nodules in lung and liver were quantified using dissecting microscopy after H&E staining. Representative H&E staining images of lungs and livers were shown.
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fig03: Knockdown of Capn4 reduces nasopharyngeal carcinoma (NPC)cell migration and invasion in vitro and in vivo. (a, b) 5-8F cells and CNE2 cells were stably transfected with constructs encoding siRNA specifically targeting Capn4 or scrambled sequence as siRNA control, after which semi-quantitative RT-PCR and Western blot analyses were performed to monitor mRNA and protein levels of Capn4 in Capn4 siRNA and control siRNA cells, as well as parental 5-8F cells and CNE2 cells. β-actin was probed as internal control. (c) Transwell assays were performed to measure in vitro migration and invasion of Capn4 siRNA and control siRNA cells. (d) To evaluate in vivo metastasis of Capn4 siRNA and control siRNA cells (n = 10 per group), 1 × 106 cells were resuspended in 0.1 mL of PBS and injected via the lateral tail vein. After 10 weeks, mice were euthanized, and metastatic nodules in lung and liver were quantified using dissecting microscopy after H&E staining. Representative H&E staining images of lungs and livers were shown.

Mentions: To determine the functional role of Capn4 in metastasis of NPC, we first generated two clones of 5-8F cells stably transfected with Capn4 siRNA (Capn4/siRNA-1 and Capn4/siRNA-2). RT-PCR and Western blot analyses demonstrated that both protein and mRNA levels of Capn4 were substantially downregulated in Capn4/siRNA-1 and Capn4/siRNA-2 cells, compared to siRNA control cells (Fig.3a,b). Then, the transwell assays were performed to assess the effects of Capn4 knockdown on capabilities of invasion and migration in NPC cells. Notably, Capn4/siRNA cells displayed significantly lower rate of invasion and migration than the parental cells and siRNA control cells (Fig.3c). In addition, MTT assays revealed that knockdown of Capn4 also inhibited cell proliferation (Fig. S1), but failed to induce apoptosis (data not shown).


Capn4 is a marker of poor clinical outcomes and promotes nasopharyngeal carcinoma metastasis via nuclear factor-κB-induced matrix metalloproteinase 2 expression.

Zheng PC, Chen X, Zhu HW, Zheng W, Mao LH, Lin C, Liu JN, Zheng M - Cancer Sci. (2014)

Knockdown of Capn4 reduces nasopharyngeal carcinoma (NPC)cell migration and invasion in vitro and in vivo. (a, b) 5-8F cells and CNE2 cells were stably transfected with constructs encoding siRNA specifically targeting Capn4 or scrambled sequence as siRNA control, after which semi-quantitative RT-PCR and Western blot analyses were performed to monitor mRNA and protein levels of Capn4 in Capn4 siRNA and control siRNA cells, as well as parental 5-8F cells and CNE2 cells. β-actin was probed as internal control. (c) Transwell assays were performed to measure in vitro migration and invasion of Capn4 siRNA and control siRNA cells. (d) To evaluate in vivo metastasis of Capn4 siRNA and control siRNA cells (n = 10 per group), 1 × 106 cells were resuspended in 0.1 mL of PBS and injected via the lateral tail vein. After 10 weeks, mice were euthanized, and metastatic nodules in lung and liver were quantified using dissecting microscopy after H&E staining. Representative H&E staining images of lungs and livers were shown.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4317905&req=5

fig03: Knockdown of Capn4 reduces nasopharyngeal carcinoma (NPC)cell migration and invasion in vitro and in vivo. (a, b) 5-8F cells and CNE2 cells were stably transfected with constructs encoding siRNA specifically targeting Capn4 or scrambled sequence as siRNA control, after which semi-quantitative RT-PCR and Western blot analyses were performed to monitor mRNA and protein levels of Capn4 in Capn4 siRNA and control siRNA cells, as well as parental 5-8F cells and CNE2 cells. β-actin was probed as internal control. (c) Transwell assays were performed to measure in vitro migration and invasion of Capn4 siRNA and control siRNA cells. (d) To evaluate in vivo metastasis of Capn4 siRNA and control siRNA cells (n = 10 per group), 1 × 106 cells were resuspended in 0.1 mL of PBS and injected via the lateral tail vein. After 10 weeks, mice were euthanized, and metastatic nodules in lung and liver were quantified using dissecting microscopy after H&E staining. Representative H&E staining images of lungs and livers were shown.
Mentions: To determine the functional role of Capn4 in metastasis of NPC, we first generated two clones of 5-8F cells stably transfected with Capn4 siRNA (Capn4/siRNA-1 and Capn4/siRNA-2). RT-PCR and Western blot analyses demonstrated that both protein and mRNA levels of Capn4 were substantially downregulated in Capn4/siRNA-1 and Capn4/siRNA-2 cells, compared to siRNA control cells (Fig.3a,b). Then, the transwell assays were performed to assess the effects of Capn4 knockdown on capabilities of invasion and migration in NPC cells. Notably, Capn4/siRNA cells displayed significantly lower rate of invasion and migration than the parental cells and siRNA control cells (Fig.3c). In addition, MTT assays revealed that knockdown of Capn4 also inhibited cell proliferation (Fig. S1), but failed to induce apoptosis (data not shown).

Bottom Line: The patients with NPC displaying higher Capn4 had a significantly shorter overall survival (P = 0.002) and progression-free survival (P = 0.003).These events resulted from Capn4 downregulation were associated with reduced expression of matrix metalloproteinase 2 (MMP2), Snail, and Vimentin.Together, these findings argue a novel function of Capn4 in invasion and metastasis of NPC, and thereby suggest that Capn4 may represent an independent prognostic factor and a potential therapeutic target in NPC.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy, School of Basic Medical Sciences, Fujian Medical University, Fuzhou, China.

Show MeSH
Related in: MedlinePlus