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Upregulation of KLHDC4 Predicts a Poor Prognosis in Human Nasopharyngeal Carcinoma.

Lian YF, Yuan J, Cui Q, Feng QS, Xu M, Bei JX, Zeng YX, Feng L - PLoS ONE (2016)

Bottom Line: Consistently, KLHDC4 knockout cell-derived xenografts also showed elevated cleaved caspase-3 and PARP but reduced Ki-67 staining.In conclusion, our results suggest that KLHDC4 promotes NPC oncogenesis by suppressing cellular apoptosis.Thus, KLHDC4 may serve as a prognosis biomarker and a potential therapeutic target for NPC.

View Article: PubMed Central - PubMed

Affiliation: Department of Experimental Research, Sun Yat-sen University Cancer Center, State Key Laboratory Oncology in South China, Collaborative Innovation Center of Cancer Medicine, Guangzhou, China.

ABSTRACT
Kelch proteins are implicated in the pathogenesis of many human diseases, including cancer. Nasopharyngeal carcinoma (NPC) is a rare malignancy in most countries, but prevalent in southern China and certain areas of Southeast Asia. In this study, we identified Kelch Domain Containing 4 (KLHDC4), an orphan member of the kelch repeat superfamily, as a prognosis marker for NPC. We examined the expression of KLHDC4 in 168 NPC cases by immunohistochemical staining and found a substantially higher level of KLHDC4 in NPC biopsies compared to adjacent normal nasopharyngeal mucosa. KLHDC4 expression was significantly related to the T classification (P <0.05), N classification (P <0.05) and total staging (P <0.01) in NPC, and patients with higher KLHDC4 expression had poorer overall (P <0.01) and metastasis-free survival (P <0.05) rates. Knockout (KO) of KLHDC4 via CRISPR/Cas9-mediated gene editing in NPC cell line dramatically inhibited cell proliferation, colony formation in soft agar and tumor formation in nude mice. In addition, cell migration and invasion were also impaired by KLHDC4 depletion as revealed by wound healing and Transwell assay. Mechanically, loss of KLHDC4 markedly induced spontaneous apoptosis in NPC cells, as evidenced by increased levels of cleaved caspase-3 and cleaved PARP. Consistently, KLHDC4 knockout cell-derived xenografts also showed elevated cleaved caspase-3 and PARP but reduced Ki-67 staining. In conclusion, our results suggest that KLHDC4 promotes NPC oncogenesis by suppressing cellular apoptosis. Thus, KLHDC4 may serve as a prognosis biomarker and a potential therapeutic target for NPC.

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Related in: MedlinePlus

Loss of KLHDC4 reduces NPC cell migration and invasion.(A) Wound-healing assays were performed at 0 and 20 hours with control and KLHDC4 KO cells. Left panels: representative images; Scale bars: 50 μm. Right panels: quantification of the wound closure area calculated by measuring the decreaseinthe wound bed surface overtime. **P <0.01. (B) KLHDC4 KO significantly reduced the invasive ability of CNE2 cells. Left panels: representative images; Scale bars: 50 μm. Right panels: quantification of average number of cells per field. ***P <0.001.
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pone.0152820.g004: Loss of KLHDC4 reduces NPC cell migration and invasion.(A) Wound-healing assays were performed at 0 and 20 hours with control and KLHDC4 KO cells. Left panels: representative images; Scale bars: 50 μm. Right panels: quantification of the wound closure area calculated by measuring the decreaseinthe wound bed surface overtime. **P <0.01. (B) KLHDC4 KO significantly reduced the invasive ability of CNE2 cells. Left panels: representative images; Scale bars: 50 μm. Right panels: quantification of average number of cells per field. ***P <0.001.

Mentions: Aberrant cell migration and invasion is frequently associated with cancer, and enhanced migration and invasion capacity is generally believed to be associated with tumor metastasis. Thus, we performed wound healing and Transwell assays to determine whether KLHDC4 expression is related to cell migration and invasion, respectively. At 20 hours after scratch injury, the KLHDC4 KO cells were able to cover only 53.3% of the scratch, whereas the control cells covered 88.0% (P <0.01; Fig 4A). Similarly, when allowing 20 hours for invasion, the parental CNE2 cells were able to invade through the Transwell chamber inserts, whereas invasion by the KLHDC4 KO cells was significantly attenuated (Fig 4B). Collectively, our results suggest that KLHDC4 deficiency dramatically impairs cell migration and invasion in vitro.


Upregulation of KLHDC4 Predicts a Poor Prognosis in Human Nasopharyngeal Carcinoma.

Lian YF, Yuan J, Cui Q, Feng QS, Xu M, Bei JX, Zeng YX, Feng L - PLoS ONE (2016)

Loss of KLHDC4 reduces NPC cell migration and invasion.(A) Wound-healing assays were performed at 0 and 20 hours with control and KLHDC4 KO cells. Left panels: representative images; Scale bars: 50 μm. Right panels: quantification of the wound closure area calculated by measuring the decreaseinthe wound bed surface overtime. **P <0.01. (B) KLHDC4 KO significantly reduced the invasive ability of CNE2 cells. Left panels: representative images; Scale bars: 50 μm. Right panels: quantification of average number of cells per field. ***P <0.001.
© Copyright Policy
Related In: Results  -  Collection

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pone.0152820.g004: Loss of KLHDC4 reduces NPC cell migration and invasion.(A) Wound-healing assays were performed at 0 and 20 hours with control and KLHDC4 KO cells. Left panels: representative images; Scale bars: 50 μm. Right panels: quantification of the wound closure area calculated by measuring the decreaseinthe wound bed surface overtime. **P <0.01. (B) KLHDC4 KO significantly reduced the invasive ability of CNE2 cells. Left panels: representative images; Scale bars: 50 μm. Right panels: quantification of average number of cells per field. ***P <0.001.
Mentions: Aberrant cell migration and invasion is frequently associated with cancer, and enhanced migration and invasion capacity is generally believed to be associated with tumor metastasis. Thus, we performed wound healing and Transwell assays to determine whether KLHDC4 expression is related to cell migration and invasion, respectively. At 20 hours after scratch injury, the KLHDC4 KO cells were able to cover only 53.3% of the scratch, whereas the control cells covered 88.0% (P <0.01; Fig 4A). Similarly, when allowing 20 hours for invasion, the parental CNE2 cells were able to invade through the Transwell chamber inserts, whereas invasion by the KLHDC4 KO cells was significantly attenuated (Fig 4B). Collectively, our results suggest that KLHDC4 deficiency dramatically impairs cell migration and invasion in vitro.

Bottom Line: Consistently, KLHDC4 knockout cell-derived xenografts also showed elevated cleaved caspase-3 and PARP but reduced Ki-67 staining.In conclusion, our results suggest that KLHDC4 promotes NPC oncogenesis by suppressing cellular apoptosis.Thus, KLHDC4 may serve as a prognosis biomarker and a potential therapeutic target for NPC.

View Article: PubMed Central - PubMed

Affiliation: Department of Experimental Research, Sun Yat-sen University Cancer Center, State Key Laboratory Oncology in South China, Collaborative Innovation Center of Cancer Medicine, Guangzhou, China.

ABSTRACT
Kelch proteins are implicated in the pathogenesis of many human diseases, including cancer. Nasopharyngeal carcinoma (NPC) is a rare malignancy in most countries, but prevalent in southern China and certain areas of Southeast Asia. In this study, we identified Kelch Domain Containing 4 (KLHDC4), an orphan member of the kelch repeat superfamily, as a prognosis marker for NPC. We examined the expression of KLHDC4 in 168 NPC cases by immunohistochemical staining and found a substantially higher level of KLHDC4 in NPC biopsies compared to adjacent normal nasopharyngeal mucosa. KLHDC4 expression was significantly related to the T classification (P <0.05), N classification (P <0.05) and total staging (P <0.01) in NPC, and patients with higher KLHDC4 expression had poorer overall (P <0.01) and metastasis-free survival (P <0.05) rates. Knockout (KO) of KLHDC4 via CRISPR/Cas9-mediated gene editing in NPC cell line dramatically inhibited cell proliferation, colony formation in soft agar and tumor formation in nude mice. In addition, cell migration and invasion were also impaired by KLHDC4 depletion as revealed by wound healing and Transwell assay. Mechanically, loss of KLHDC4 markedly induced spontaneous apoptosis in NPC cells, as evidenced by increased levels of cleaved caspase-3 and cleaved PARP. Consistently, KLHDC4 knockout cell-derived xenografts also showed elevated cleaved caspase-3 and PARP but reduced Ki-67 staining. In conclusion, our results suggest that KLHDC4 promotes NPC oncogenesis by suppressing cellular apoptosis. Thus, KLHDC4 may serve as a prognosis biomarker and a potential therapeutic target for NPC.

Show MeSH
Related in: MedlinePlus