Downregulation of DENND2D by promoter hypermethylation is associated with early recurrence of hepatocellular carcinoma

Kanda, Mitsuro; Nomoto, Shuji; Oya, Hisaharu; Takami, Hideki; Hibino, Susumu; Hishida, Mitsuhiro; Suenaga, Masaya; Yamada, Suguru; Inokawa, Yoshikuni; Nishikawa, Yoko; Asai, Mikako; Fujii, Tsutomu; Sugimoto, Hiroyuki; Kodera, Yasuhiro

doi:10.3892/ijo.2013.2165

Cited by 40 publications

(37 citation statements)

References 33 publications

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“…Methylation-specific PCR (MSP) and bisulfite sequence analysis DENND2D possesses a CpG island around the promoter region [24], and we hypothesized that aberrant methylation is responsible for regulation of DENND2D transcription in GC. DNA samples from 12 GC cell lines and 112 GC tissues and corresponding noncancerous tissues were Expression of DENND2D in GC 289 treated with bisulfite, and conventional MSP was performed.…”

Section: Reverse-transcription Polymerase Chain Reaction (Rt-pcr) Andmentioning

confidence: 99%

“…Total RNA (10 lg per sample) was isolated from 12 GC cell lines and 112 primary GC tissues and corresponding noncancerous tissues and was used to generate complementary DNA. PCR primers and conditions for RT-PCR and qRT-PCR were as described previously [24]. qRT-PCR was performed using 12 GC cell lines and 112 pairs of clinical samples in triplicate and included no-template samples as negative controls.…”

Section: Reverse-transcription Polymerase Chain Reaction (Rt-pcr) Andmentioning

confidence: 99%

“…Membrane trafficking proteins are assumed to have close implications for drug delivery, which is important for GC treatment [21,22]; however, the role and regulatory mechanisms of DENND family proteins in GC have not been reported. We recently took particular note of DENND2D, which is located on chromosome 1p13.3 and encodes a 53-kDa protein [23], and reported that DENND2D has tumor suppressive activity in hepatocellular carcinoma [24]. Accordingly, we investigated the expression and regulatory mechanism of DEN-ND2D to identify a novel biomarker for GC.…”

Section: Introductionmentioning

confidence: 99%

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Prognostic impact of expression and methylation status of DENN/MADD domain-containing protein 2D in gastric cancer

et al. 2014

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Background Patients with advanced gastric cancer (GC) have an adverse prognosis even after curative resection. Development of novel diagnostic and therapeutic approaches for GC is urgently required. Methods The expression and methylation status of DENN/MADD domain-containing protein 2D (DEN-ND2D), a member of the membrane trafficking proteins, were evaluated in 12 GC cell lines and 112 pairs of surgical specimens. Subgroup analysis based on tumor differentiation, location, and morphology was also performed. Expression and distribution of DENND2D protein were determined by immunohistochemistry. Results The majority of GC cell lines (75 %) and tissues (79 %) showed reduced expression of DENND2D mRNA compared with noncancerous gastric tissues. GC tissues showed a significantly lower mean expression level of mRNA and a higher frequency of promoter hypermethylation of DENND2D than corresponding noncancerous tissues. No significant differences in DENND2D mRNA expression and methylation status were found between GC subtypes categorized by tumor differentiation, location, and morphology. The expression patterns of DENND2D protein were confirmed to be consistent with those of DENND2D mRNA. Downregulation of DENND2D mRNA in GC tissues was significantly associated with factors related to more advanced GC and subsequent adverse prognosis. Among 72 patients who underwent R0 resection, downregulation of DENND2D mRNA in GC tissues was an independent prognostic factor and associated with early recurrence. Conclusions Our results suggested that DENND2D is a putative tumor suppressor gene regulated by promoter hypermethylation in GC. Downregulation of DENND2D can serve as a novel tumor biomarker to predict progression and early recurrence of all types of GC.

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Section: Reverse-transcription Polymerase Chain Reaction (Rt-pcr) Andmentioning

confidence: 99%

Section: Reverse-transcription Polymerase Chain Reaction (Rt-pcr) Andmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Prognostic impact of expression and methylation status of DENN/MADD domain-containing protein 2D in gastric cancer

et al. 2014

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“…The intensity and pattern of SAMSN1 protein expression was determined by immunohistochemical staining using 48 representative sections of well-preserved GC tissue as described previously (25). Sections were incubated for 1 h at room temperature with a rabbit polyclonal antibody raised against SAMSN1 (catalog no., 13063-1-AP; ProteinTech Group, Inc., Chicago, IL, USA) diluted 1:400 in antibody diluent (Dako, Glostrup, Denmark).…”

Section: Cellmentioning

confidence: 99%

Prognostic relevance of SAMSN1 expression in gastric cancer

et al. 2016

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Abstract.The prognosis for patients with advanced gastric cancer (GC) remains poor. The identification of biomarkers relevant to the recurrence and metastasis of GC is advantageous for stratifying patients and proposing novel molecular targets. In the present study the oncological roles of SAM domain, SH3 domain and nuclear localization signals 1 (SAMSN1), a mediator of B-cell function, were elucidated in GC. The expression and methylation status of SAMSN1 were investigated in a panel of 11 GC cell lines. Immunohistochemical staining was performed to determine the pattern of SAMSN1 protein expression in gastric tissues. The prognostic impact of SAMSN1 expression was determined by analyzing 175 pairs of surgically resected gastric tissues. A marked decrease in the level of SAMSN1 mRNA was detected in 8/11 GC cell lines as compared with that in a non-transformed intestinal epithelium cell line (FHs 74) without promoter methylation. The mean expression level of SAMSN1 mRNA was reduced in GC tissues compared with normal adjacent tissues, an observation that was independent of tumor differentiation. The pattern of SAMSN1 protein expression was significantly correlated with that of SAMSN1 mRNA. Low SAMSN1 mRNA expression was significantly associated with tumor size (>60 mm; P=0.026) and shorter overall survival times (P=0.004). Multivariate analysis identified low SAMSN1 mRNA expression as an independent prognostic factor for poor overall survival (hazard ratio, 1.80; 95% confidence interval, 1.07-3.05; P= 0.025). The difference in survival between the low and high SAMSN1 expression groups was more marked in patients with stage II/III GC compared to those with stage IV GC. In patients with stage II/III GC who underwent curative surgery, low SAMSN1 expression was associated with reduced disease free survival times. The results of the present study indicate that downregulation of SAMSN1 transcription may affect the progression and recurrence of GC, and therefore may represent a novel biomarker of GC.

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“…Since epigenetic alterations in cancer cells affect virtually every cellular pathway such as those involved in cell-cycle progression, angiogenesis, apoptosis, cell survival and immunogenicity, it is thought that epigenetic drugs will possess versatile antitumor activity (18,20,21). In addition, abnormal hypermethylated genes in cancer can serve as biomarkers for early detection and tumor classification, and for monitoring response to treatments such as targeted therapy and epigenetic agents (22)(23)(24).…”

Section: Introductionmentioning

confidence: 99%

Identification of intragenic methylation in the TUSC1 gene as a novel prognostic marker of hepatocellular carcinoma

et al. 2013

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Abstract.Patients with hepatocellular carcinoma (HCC) have a poor prognosis, and novel molecular targets for treating recurrence and progression of the disease along with associated biomarkers are urgently required. In the present study, expression and the regulatory mechanism of TUSC1 (tumor suppressor candidate 1) were investigated to determine if it is a candidate tumor suppressor gene for HCC, which shows repressed transcription that involves aberrant DNA methylation. TUSC1 mRNA expression levels in HCC cell lines and 94 pairs of surgical specimens were determined using quantitative real-time reverse transcription polymerase chain reaction assay. Methylation status of HCC cell lines and clinical samples were analyzed to investigate the regulatory mechanism of TUSC1 transcription and the relationship between the methylation status of the TUSC1 gene and clinicopathological factors. The expression and distribution of the TUSC1 protein in liver tissues were determined using immunohistochemistry. A majority of HCC cell lines (89%) and surgical specimens (84%) demonstrated reduced expression levels of TUSC1 mRNA compared with paired non-cancerous liver tissues. The mean mRNA expression level in HCC was significantly lower than in corresponding non-cancerous liver. In contrast, no significant difference was found in TUSC1 mRNA expression level between adjacent normal and cirrhotic liver tissue from HCC patients. The TUSC1 protein expression pattern in HCC and liver tissues was consistent with TUSC1 mRNA expression. Twenty-nine (31%) of 94 patients showed intragenic hypermethylation of the TUSC1 gene in HCC, and hypermethylation was significantly associated with advanced pathological stage. Subsequently, patients with hypermethylation of the TUSC1 gene had a significantly poorer prognosis than patients without hypermethylation. Our results suggest that TUSC1 is a candidate tumor suppressor gene and intragenic hypermethylation is one of the suppressive mechanisms that regulate TUSC1 transcription in HCC. Intragenic methylation of the TUSC1 gene may serve as a novel prognostic marker of HCC.

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Downregulation of DENND2D by promoter hypermethylation is associated with early recurrence of hepatocellular carcinoma

Cited by 40 publications

References 33 publications

Prognostic impact of expression and methylation status of DENN/MADD domain-containing protein 2D in gastric cancer

Prognostic impact of expression and methylation status of DENN/MADD domain-containing protein 2D in gastric cancer

Prognostic relevance of SAMSN1 expression in gastric cancer

Identification of intragenic methylation in the TUSC1 gene as a novel prognostic marker of hepatocellular carcinoma

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