Proteomics of the Radioresistant Phenotype in Head-and-Neck Cancer: Gp96 as a Novel Prediction Marker and Sensitizing Target for Radiotherapy

Lin, Tzu‐En; Chang, Joseph Tung-Chieh; Wang, Hung‐Ming; Chan, Shih-Hsuan; Chiu, Ching-Chi; Lin, Chien‐Yu; Fan, Kang‐Hsing; Liao, Chun‐Ta; Chen, I-How; Liu, Tsan Z.; Li, Hsiao-Fang; Cheng, Ann‐Joy

doi:10.1016/j.ijrobp.2010.03.002

Cited by 47 publications

(50 citation statements)

References 34 publications

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“…To suppress miR-196 expression, 300 μM antagomir oligonucleotides were transfected into the cells. Transfection was performed using the Lipofectamine 2000™ reagent (Invitrogen) in OPTI-MEM medium (Invitrogen), and the cells were incubated at 37°C in a humidified atmosphere with 5% CO 2 for 10 h, similarly as previously described [22]. Afterward, the medium was replaced with fresh complete medium, and the cells were continuously cultured.…”

Section: Materials Subjects and Methodsmentioning

confidence: 99%

OncomiR-196 promotes an invasive phenotype in oral cancer through the NME4-JNK-TIMP1-MMP signaling pathway

Chang

Liao

et al. 2014

Mol Cancer

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Section: Materials Subjects and Methodsmentioning

confidence: 99%

OncomiR-196 promotes an invasive phenotype in oral cancer through the NME4-JNK-TIMP1-MMP signaling pathway

Chang

Liao

et al. 2014

Mol Cancer

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“…31 Briefly, a total of 100 cells was seeded into a six-well plate and irradiated with serial doses of radiation (0-8 Gy). The cells were cultured for 7 days to allow colony formation.…”

Section: Western Blot Analysismentioning

confidence: 99%

Grp78 as a therapeutic target for refractory head–neck cancer with CD24−CD44+ stemness phenotype

Chiu

Lee

et al. 2013

Cancer Gene Ther

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Cancer stem cells are refractory to conventional therapy, which result to cancer metastasis and chemo-radioresistance. Grp78 is known to have important roles in cytoprotection and tumorigenesis in several cancers. We therefore examined whether Grp78 can serve as a therapeutic target for refractory stemness phenotype of head and neck cancer (HNC). Six HNC cell lines were used. Fluorescence-activated cell sorting (FACS) analysis was used to sort CD24(-)CD44(+) and Grp78(+) cells. The small interfering RNA (siRNA) knockdown and cDNA transfection were applied to examine the effects of Grp78 on cellular function. Western blot and confocol microscopy were used to determine the effects of downstream protein expressions. Xenografted mouse tumors and immunohistochemistry were used to validate the results. We found that Grp78 regulated the conversion of CD24(-)CD44(+) cells, a characteristic of HNC stem cells. The CD24(-)CD44(+)Grp78(+) cells showed superior chemo-radioresistance and invasion ability compared with CD24(-)CD44(+), Grp78(+) or the parental cells. Silencing Grp78 increased chemo-radiosensitivity, inhibited cell invasion, reverse epithelial-mesenchymal transition, suppressed cancer stemness, withdrew CD24(-)CD44(+) cell conversion and induced differentiated phenotype. Study in xenografted mice further showed that CD24(-)CD44(+)Grp78(+) cells exhibited highest tumorigenesis, compared with CD24(-)CD44(+) CD24(+)CD44(+) or the parental cells. Grp78 knockdown dramatically restrained tumor growth along with the inhibition of stem cell regulatory proteins Oct-4 and Slug. Grp78 may serve as a molecular target that can be further developed for eradication of refractory HNC with stemness phenotype.

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“…To better identify cell proteomes, subcellular proteins were extracted as previously described (13). Fractions containing 50 mg of protein were separated by electrophoresis on an 8% to 16% gradient gel.…”

Section: Subcellular Protein Extraction and Comparative Proteomic Anamentioning

confidence: 99%

Molecular Chaperones as a Common Set of Proteins That Regulate the Invasion Phenotype of Head and Neck Cancer

Chiu

Lin

Lee

et al. 2011

Clinical Cancer Research

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Purpose: The goal of this study was to establish a common set of molecules that regulate cell invasion in head and neck cancer (HNC).Experimental Design: Five invasive sublines derived from HNC cell lines were established using the Matrigel selection method. Proteomic technology, MetaCore algorithm, and reverse transcriptase-PCR methods were used to search for molecules that contribute to the invasion phenotype. Cellular functional analyses and clinical association studies were applied to examine the significance of the molecules.Results: Fifty-two proteins were identified in more than two of the four independent proteomic experiments, including 10 (19%) molecular chaperones. Seven chaperones were confirmed to be differentially expressed in five sublines, Hsp90a, Hsp90b, Hsp90-B1/Gp96, Hsp70-A5/Grp78, and HYOU1, that upregulate, whereas Hsp60 and glucosidase-a neutral AB (GANAB) downregulate. Four molecules were further investigated. In all cell lines, knockdown of Hsp60 or GANAB and silencing of Gp96 or Grp78 considerably enhanced or reduced cell migration and invasion, respectively. Clinical association studies consistently revealed that low levels of Hsp60 or GANAB and high levels of Gp96 or Grp78 are significantly associated with advanced cancer (P < 0.001 to P ¼ 0.047, respectively, for the four molecules) and poor survival (P < 0.001 to P ¼ 0.025, respectively, for the four molecules).Conclusion: Our study defined molecular chaperones as a common set of proteins that regulate the invasion phenotype of HNC. Loss of the tumor suppression function of Hsp60 or GANAB and acquisition of the oncogenic function of Gp96 or Grp78 contribute to aggressive cancers. These molecules may serve as prognostic markers and targets for cancer drug development.

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Proteomics of the Radioresistant Phenotype in Head-and-Neck Cancer: Gp96 as a Novel Prediction Marker and Sensitizing Target for Radiotherapy

Cited by 47 publications

References 34 publications

OncomiR-196 promotes an invasive phenotype in oral cancer through the NME4-JNK-TIMP1-MMP signaling pathway

OncomiR-196 promotes an invasive phenotype in oral cancer through the NME4-JNK-TIMP1-MMP signaling pathway

Grp78 as a therapeutic target for refractory head–neck cancer with CD24−CD44+ stemness phenotype

Molecular Chaperones as a Common Set of Proteins That Regulate the Invasion Phenotype of Head and Neck Cancer

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