Silencing HOXD10 by promoter region hypermethylation activates ERK signaling in hepatocellular carcinoma

Guo, Yulin; Peng, Yawen; Gao, Dan; Zhang, Meiying; Yang, Weili; Linghu, Enqiang; Herman, James; Fuks, François; Dong, Guanglong; Guo, Mingzhou

doi:10.1186/s13148-017-0412-9

Cited by 38 publications

(44 citation statements)

References 31 publications

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“…All these cell lines were preserved in our institute (The First Medical Centre, Chinese PLA General Hospital, Beijing, China) and were cultured in RPMI 1640 supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin. Semi-quantitative RT-PCR to evaluate mRNA expression of CD40 in colon cancer cell lines with or without 5-aza-2'-deoxycytidine (5-Aza, Sigma) treatment (2 mM for 96h) were carried out as previously described [30]. Genomic DNA was prepared by the proteinase K method.…”

Section: Experimental Validation In Colon Cancer Cell Linesmentioning

confidence: 99%

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Identification of a Methylation-Driven Gene Panel for Survival Prediction in Colon Cancer

Peng

Zhao

Yin

et al. 2020

Preprint

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Background: Prediction and improvement of prognosis is important for effective clinical management of colon cancer patients. Accumulation of a variety of genetic as well as epigenetic changes in colon epithelial cells has been identified as one of the fundamental processes that drive the initiation and progression of colon cancer. This study aimed to explore functional genes regulated by DNA methylation and the potential of these DNA methylation changes to become biomarkers predictive of colon cancer prognosis.Methods: Methylation-driven genes (MDGs) were explored by applying an integrative analysis tool (MethylMix) to The Cancer Genome Atlas (TCGA) colon cancer project. TCGA colon cancer patients with available survival information (n=281) were randomly divided into training dataset (50%) for model construction and testing dataset (50%) for model validation. The prognostic MDG panel was identified in the training dataset by combining the Cox regression model with the least absolute shrinkage and selection operator regularization, a widely used approach to penalize the effect of multicollineatity. GSEA was employed to determine functional pathways associated with the prognostic 6-MDG panel. CD40 expression and methylation in colon cancer samples were also examined in datasets (expression profile [GSE8671] and methylation profile [GSE42752]) from Gene Expression Omnibus. Experimental confirmation of DNA methylation in colon cancer cell lines was performed using methylation specific PCR and bisulfite sequencing.Results: We identified and internal validated a prognostic methylation-driven gene panel consisting of six gene members (TMEM88, HOXB2, FGD1, TOGARAM1, ARHGDIB and CD40). High risk phenotype classified by the 6-MDGs panel was associated with cancer-related biological processes, including invasion and metastasis, angiogenesis and tumor immune microenvironment, among others. The prognostic value of the 6-MDGs panel was independent of TNM stage, and its combination with TNM stage and age could help improve survival prediction of colon cancer patients. Additionally, we validated that the expression of CD40 was regulated by promoter region methylation in colon cancer samples and cell lines. Conclusions: The proposed 6-MDGs panel represents a promising signature for estimating overall survival in patients with colon cancer.

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Section: Experimental Validation In Colon Cancer Cell Linesmentioning

confidence: 99%

“…Total protein of CD40 in these colon cancer cell lines was measured by western blot, which was performed as previously described [30]. b-actin was used as a loading control.…”

Section: Experimental Validation In Colon Cancer Cell Linesmentioning

confidence: 99%

Identification of a Methylation-Driven Gene Panel for Survival Prediction in Colon Cancer

Peng

Zhao

Yin

et al. 2020

Preprint

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“…Hypermethylation of promoters of a number of TSGs, such as EGF-containing fibulin extracellular matrix protein 1, glutathione S-transferase P1, suppressor of cytokine signaling 3, 3OST2, basic helix-loop-helix family member E22/cysteine deoxygenase 1/CUGBP Elva-like family member 4, SHP1 and transmembrane protein with EGF-like and two follistatin-like domains 2, have been studied in certain EC tissues and cell lines (such as Ishikawa and KLE) and the methylation frequency is high (26)(27)(28)(29)(30)(31). The present study investigated five TSGs (HOXD10, SHH, ZNF545, PCDH17 and MEIS1) whose promoters are reported to be hypermethylated in other malignancies but have not been evaluated for methylation in EC (32)(33)(34)(35)(36)(37)(38)(39)(40). For example, HOXD10 is hypermethylated and lowly expressed in colon adenocarcinoma cells, which downregulates the RHOC/AKT/MAPK pathway to enhance apoptosis and restrict the proliferation, migration and invasion of colon adenocarcinoma (32,33).…”

Section: Introductionmentioning

confidence: 99%

“…The present study investigated five TSGs (HOXD10, SHH, ZNF545, PCDH17 and MEIS1) whose promoters are reported to be hypermethylated in other malignancies but have not been evaluated for methylation in EC (32)(33)(34)(35)(36)(37)(38)(39)(40). For example, HOXD10 is hypermethylated and lowly expressed in colon adenocarcinoma cells, which downregulates the RHOC/AKT/MAPK pathway to enhance apoptosis and restrict the proliferation, migration and invasion of colon adenocarcinoma (32,33). Methylation of the Shh promoter and reduced expression of SHH is frequent in basal cell carcinoma (34,35).…”

Section: Introductionmentioning

confidence: 99%

Frequent promoter methylation of HOXD10 in endometrial carcinoma and its pathological significance

et al. 2020

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Homeobox D 10 (HOXD10) is important in cell differentiation and morphogenesis and serves as a tumor suppressor gene (TSG) in a number of malignancies. The present study investigated its promoter methylation status and association with the clinicopathological features of endometrial cancer (EC), and measured HOXD10 protein expression levels. EC samples (n=62), including 50 endometroid adenocarcinoma (EA) and 12 mucinous endometrial carcinoma samples (EC) and 70 non-cancerous samples were collected. All samples were evaluated for the methylation status of several TSGs, including HOXD10, using methylation-specific PCR. HOXD10 expression level was evaluated using immunohistochemistry. 5-Aza-2-deoxycytidine treatment was performed in the EC cell line Ishikawa to observe the change in HOXD10 expression levels. HOXD10 promoter methylation was more frequent in cancer samples (P<0.001). Downregulation of HOXD10 in EC samples was confirmed at the protein level using immunohistochemistry (P<0.001) and immunohistochemical staining was negatively associated with methylation status (P<0.05). Less HOXD10 protein was expressed in MEC compared with EA samples (P<0.001). The HOXD10 promoter was hypermethylated in both EA and MEC, causing decreased HOXD10 protein expression levels in EC cells. HOXD10 expression levels were partially reversed by 5-Aza-2-deoxycytidine treatment. The results of the present study demonstrated that epigenetic silencing of HOXD10 putatively contributed to the tumorigenesis of EA. Although there was no significant difference in HOXD10 methylation between EA and MEC, HOXD10 protein expression levels differed between these two diseases, indicating that it may be a useful protein biomarker for distinguishing between these two lesions.

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“…1 Various risk factors, such as chronic infections with hepatitis B or C virus and aflatoxins, may contribute to HCC pathogenesis. 1 Various risk factors, such as chronic infections with hepatitis B or C virus and aflatoxins, may contribute to HCC pathogenesis.…”

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confidence: 99%

Inhibition of cyclin‐dependent kinase 7 suppresses human hepatocellular carcinoma by inducing apoptosis

Zhong

Yang

Jia

et al. 2018

J of Cellular Biochemistry

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Increasing evidence has shown that THZ1, a covalent cyclin-dependent kinase 7 (CDK7) inhibitor, exhibits therapeutic effects in various tumors. However, the possible effect of THZ1 on hepatocellular carcinoma (HCC) remains unknown. Our study was to investigate the roles of THZ1 in HCC cells and in subcutaneous HCC model and illustrate the molecular mechanisms. The phosphorylation levels of Ser2, Ser5, and Ser7 within RNA polymerase II (RNAPII) C-terminal domain (CTD) and the expression levels of Ki67, Mcl-1, survivin, XIAP, and p53 in HCC cells under different conditions were examined by Western blot analysis. Cell growth and apoptosis were assessed via 3-(4,5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide (MTT) assay and flow cytometry analysis, respectively. Tumor volume was assessed in HCC mice with THZ1 or vehicle treatment and immunohistochemical (IHC) analysis was conducted on excised tumors. THZ1 significantly inhibited the phosphorylation of Ser2, Ser5, and Ser7 within RNAPII-CTD in the dose-dependent and irreversible manner. MTT assay and flow cytometry analysis showed that THZ1 inhibited HCC cell proliferation and induced apoptosis, respectively. Western blot analysis indicated THZ1 significantly upregulated p53 expression and downregulated the expressions of Mcl-1, survivin, XIAP, and Ki67. THZ1 suppressed tumor growth in Hep3B xenografted mice in a time-dependent manner. IHC analysis indicated that tumors in THZ1 group had less Ki67+ cells and more cleaved caspase-3+ cells than those in vehicle group. THZ1 exhibited anti-HCC effects through irreversibly inhibiting CDK7 activity, decreasing RNAPII-CTD phosphorylation, inducing p53 expression and inhibiting antiapoptotic gene expressions, which subsequently induced apoptosis and inhibited proliferation of HCC cells.

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Silencing HOXD10 by promoter region hypermethylation activates ERK signaling in hepatocellular carcinoma

Cited by 38 publications

References 31 publications

Identification of a Methylation-Driven Gene Panel for Survival Prediction in Colon Cancer

Identification of a Methylation-Driven Gene Panel for Survival Prediction in Colon Cancer

Frequent promoter methylation of HOXD10 in endometrial carcinoma and its pathological significance

Inhibition of cyclin‐dependent kinase 7 suppresses human hepatocellular carcinoma by inducing apoptosis

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