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To investigate the methylation of 5'-CpG island and expression of RUNX3 in salivary gland adenoid cystic carcinoma cell lines. RT-PCR (reverse transcription-polymerase chain reaction), laser scanning confocal microscope (LSCM) and Western blot were used to detect the expression of RUNX3 gene and protein in salivary gland adenoid cystic carcinoma cell lines, ACC-2, ACC-3, and ACC-M, before/after a treatment of 5-Aza-dc respectively. A weak expression of RUNX3 was found in ACC-2 and ACC-3. And no expression of RUNX3 was found in ACC-3 cell line. After a treatment of 300 nmol/L 5-Aza-dc for 72 hours, the expression of RUNX3 in ACC-2 and ACC-3 cells was enhanced, and in ACC-M was restored. LSCM results showed that the RUNX3 protein was located mainly in the cytoplasm of ACC cell lines. After a treatment of 300 nmol/L 5-Aza-dc for 72 h, both nuclear and cytoplasmic location of RUNX3 positive signals were found in the ACC-2 and ACC-3 cells. However, a weak positive signal was still only found in the cytoplasm of ACC-M cells. Partial methylation in promoter 5'-CpG island of RUNX3 gene was found in all three cell lines. And the methylation degree of CpG island was 50%, 75% and 33% in ACC-2, ACC-M and ACC-3 respectively. After a treatment of 5-Aza-dc, the RUNX3 gene showed unmethylated status in all three cell lines. The methylation of RUNX3 plays an important role in the silencing of RUNX3 expression in ACC cell lines. The cytoplasmic mislocalization of RUNX3 may be correlated with the inhibition of its function in ACC cells. Source


Tan Z.,Zhejiang Province Cancer Hospital
Zhonghua zhong liu za zhi [Chinese journal of oncology] | Year: 2010

To investigate the evolution pattern of the Runx3 gene 5'-CpG island ~3478 bp region methylation in human salivary gland adenoid cystic carcinoma (SGACC). Quantitative MSP method was used to detect the methylation status of CpG island in various regions (No.1-10) of Runx3 promoter region, and Western blot was used for detection of the expression of Runx3 protein in 41 salivary gland SGACC samples and corresponding non-neoplastic salivary gland tissues. A Logistic model was used to analyze the risk ratio between the methylation status of CpG island in Runx3 gene and development of salivary SGACC, meanwhile, the possible association among the methylation of Runx3 gene, the clinicopathological parameters of SGACCs, and Runx3 protein expression was compared. The results of qMSP showed that the hypermethylation initially occurred at the most 5' region of the Runx3 CpG island and spread to the transcription start site. The methylation rate was highest in region No. 1 and No. 2 among the successive ten regions ranging from the 5' region to the transcription start site within the Runx3 CpG island, and lowest in the transcription start site both in SGACCs and normal salivary glands. Furthermore, there was no methylation in the transcription start site in nomal salivary glands tissues. Together with the results of Logistic model analysis, those results indicate that the transcription start site within the Runx3 promoter CpG island is critical for gene silencing. Western blot results revealed that the Runx3 protein level in SGACC was significantly lower than that in normal salivary glands (P < 0.01). In combination of the results of qMSP, it is presumed that the Runx3 gene methylation is one of the reason inducing the down-regulation of Runx3 in SGACCs. Methylation of the Runx3 CpG island spreads from the most 5'-region to the transcription start site in human salivary gland adenoid cystic carcinoma, and the transcription start site may be a critical region for the methylation of Runx3. The evolution pattern of Runx3 gene methylation is related to the tumorigenesis of SGACCs. Source


Ge M.-H.,Zhejiang Province Cancer Hospital
Chinese Journal of Oncology | Year: 2012

Objective: To assess the epidermal growth factor receptor (EGFR) status in salivary adenoid cystic carcinoma and explore its role in cancer invasion. Methods: Fifty-four patients with pathologically confirmed salivary adenoid cystic carcinoma (SACC) were divided into invasion group and non-invasion group. The EGFR expression was determined by immunohistochemstry (SP staining). The relations between the EGFR expression and the SACC clinical pathological characteristics were analyzed. Results: EGFR were mainly expressed in the cell membrane and cytoplasm in the tissue of SACC. The positive rate of EGFR expression in the tumor tissue was 75.9% (41/54), and EGFR was over-expressed in the cytoplasm. The positive rate of EGFR expression in invasion group was higher than that in the non-invasion group (10.0%, P <0.05). EGFR expression were related with the SACC T stages, histological types, distant metastasis, lymph node metastasis, and nerve invasion (P < 0.05). Conclusions: A higher expression of EGFR gene in the cytoplasm may have important effect on the progression of invasive carcinoma. Further investigations are required to develop new strategy in the treatment of salivary adenoid cystic carcinoma. Source


Ling Z.Q.,Zhejiang Cancer Research Institute | Zhao Q.,Zhejiang Province Cancer Hospital | Zhou S.L.,Zhejiang Cancer Research Institute | Mao W.M.,Zhejiang Cancer Research Institute
European Journal of Surgical Oncology | Year: 2012

Background: Tumor-specific alterations of DNA methylation in circulating DNA have been associated with tumor burden and malignant progression. A wealth of information indicating the potential use of DNA methylation in circulating DNA for cancer screening, prognosis and monitoring of the efficacy of anticancer therapies has emerged. In this study, we examined prospectively whether the presence of plasma DNA with tumor characteristics before oesophagectomy is a predictive factor related to disease-free survival (DFS). Methods: Promoter hypermethylation of MSH2 was analyzed using real-time methylation-specific PCR (real-time MSP) in paired tumor and plasma samples of 209 patients with esophageal squamous cell carcinoma (ESCC). Results: Aberrant MSH2 methylation was found in 101 of 209 ESCC patients. Of these 101 patients, 77 cases exhibited the same alteration in their plasma DNA. No alterations were found in the plasma DNA of the remaining 108 patients. As a control, we screened for aberrant methylation in the plasma DNA of 60 health individuals. No methylation was found in plasma DNA of these control groups. Follow-up analysis indicated significantly lower DFS for patients with high MSH2 methylation compared to those with MSH2 unmethylation after surgery. Conclusions: It was suggestted that MSH2 methylation in the plasma would be a good predictor of DFS for these ESCC patients before oesophagectomy. © 2011 Elsevier Ltd. All rights reserved. Source


Yang H.,Chemotherapy Center | Chen Z.,Laboratory of Clinical Pharmacy | Wu M.,Zhejiang Province Cancer Hospital | Lei T.,Chemotherapy Center | And 2 more authors.
Cancer Biology and Therapy | Year: 2016

Background: Poorly Differentiated Thyroid Carcinoma (PDTC), especially advanced PDTC, is an aggressive disease and displays a much poorer prognosis compared with well differentiated thyroid carcinoma. Surgery is the recommended treatment in the early stage of PDTC, however, no effective treatment modalities are currently available for advanced PDTCMethods: Two advanced PDTC patients with no radioiodine uptake adopted a cytotoxic chemotherapy with liposomal doxorubicin (35 mg/m2, day 1) plus cisplatin (75 mg/m2, day1–3) every 3 weeks. Computer tomography (CT) was performed after 6 cycles (case 1) or 5 cycles (case 2) of chemotherapyResults: Our patients achieved remarkable response with one a Complete Remission (CR) and the other a very good Partial Remission (PR)Conclusion: Our findings indicate that liposomal doxorubicin-based chemotherapy regimens might produce response in PDTC patients, and improve their overall survival and quality of life. Hence we believe this result is very important for oncologists in treating PDTC. © 2016 Taylor & Francis Group, LLC. Source

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