DCLK1基因启动子甲基化与口腔鳞癌临床病理特征及预后的关系
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摘要
目的探讨双肾上腺素样激酶1(doublecortin-like kinase 1,DCLK1)基因启动子甲基化与口腔鳞癌临床病理特征及预后的关系。方法选取手术切除的129例口腔鳞癌组织和对应癌旁组织,采用甲基化特异性PCR检测DCLK1基因启动子甲基化情况,分析其与临床病理特征之间的关系。RT-PCR和免疫组织化学法分别检测DCLK1 mRNA和蛋白表达水平,并分析DCLK1基因启动子甲基化与DCLK1 mRNA和蛋白表达之间关系。随访3年,采用Kaplan-Meier法绘制生存曲线,分析DCLK1基因启动子甲基化对患者预后的影响。结果口腔鳞癌组织和癌旁组织中DCLK1基因启动子甲基化阳性表达率分别为35. 7%和64. 3%(P <0. 01)。口腔鳞癌组织中DCLK1mRNA水平和蛋白表达均较癌旁组织上升(均P <0. 01)。口腔鳞癌组织中DCLK1基因启动子甲基化阳性表达与DCLK1 mRNA和蛋白表达之间均呈负相关(r=-0. 767,P <0. 01; r=-0. 671,P <0. 01)。DCLK1基因启动子甲基化状态在口腔鳞癌患者TNM分期、组织分化程度和淋巴结转移方面比较,差异均具有统计学意义(均P <0. 05)。DCLK1基因启动子甲基化阳性表达患者3年无进展生存率和总生存率均高于阴性表达患者(63. 0%vs 43. 4%,69. 6%vs 47. 0%,均P <0. 05)。结论 DCLK1去甲基化表达异常,与口腔鳞癌临床病理特征相关,影响患者预后。DCLK1去甲基化状态可能通过参与调控DCLK1 mRNA表达和蛋白分泌在肿瘤发生和发展发挥作用。
Objective To investigate the relationship of double cortin-like kinase 1( DCLK1) gene promoter methylation with clinicopathological features and prognosis of patients with oral squamous cell carcinoma. Methods One-hundredand-twenty-nine patients with oral squamous cell carcinoma were enrolled in the study. Methylation-specific PCR was used to detect the methylation status of DCLK1 gene promoter in cancer tissues and corresponding pericancerous tissues,and its relationship with clinicopathological features was analyzed. The expression levels of DCLK1 mRNA and protein were detected by RT-PCR and immunohistochemistry,respectively,and the relationship between DCLK1 promoter methylation and DCLK1 expression was analyzed. The survival curve was drawn by Kaplan-Meier method after 3-year follow-up,and the correlation of DCLK1 gene promoter methylation with survival of patients was analyzed. Results The positive expression rates of DCLK1 gene promoter methylation in oral squamous cell carcinoma tissues and pericancerous tissues was 35. 7%and 64. 3%,respectively( P < 0. 01). The expression of DCLK1 mRNA and protein in oral squamous cell carcinoma was higher than that in pericancerous tissues( both P < 0. 01). The DCLK1 gene promoter methylation was negatively correlated with the expressions of DCLK1 mRNA and protein in oral squamous cell carcinoma( r =-0. 767,P < 0. 01; r =-0. 671,P < 0. 01). The methylation status of DCLK1 gene promoter was significantly associated with TNM stage,histological differentiation,and lymph node metastasis in patients with oral squamous cell carcinoma( all P < 0. 05). The 3-year progression-free survival and overall survival of patients with positive DCLK1 gene promoter methylation were higher than those of negative DCLK1 gene promoter methylation( 63. 0% vs 43. 4%,69. 6% vs 47. 0%,both P < 0. 05). Conclusion The methylation status of DCLK1 gene promoter is related to clinicopathological features of oral squamous cell carcinoma,and can affect the prognosis of patients. DCLK1 demethylation may play a role in the occurrence and development of tumor by being involved in the regulation of DCLK1 mRNA expression and protein secretion.
Objective To investigate the relationship of double cortin-like kinase 1( DCLK1) gene promoter methylation with clinicopathological features and prognosis of patients with oral squamous cell carcinoma. Methods One-hundredand-twenty-nine patients with oral squamous cell carcinoma were enrolled in the study. Methylation-specific PCR was used to detect the methylation status of DCLK1 gene promoter in cancer tissues and corresponding pericancerous tissues,and its relationship with clinicopathological features was analyzed. The expression levels of DCLK1 mRNA and protein were detected by RT-PCR and immunohistochemistry,respectively,and the relationship between DCLK1 promoter methylation and DCLK1 expression was analyzed. The survival curve was drawn by Kaplan-Meier method after 3-year follow-up,and the correlation of DCLK1 gene promoter methylation with survival of patients was analyzed. Results The positive expression rates of DCLK1 gene promoter methylation in oral squamous cell carcinoma tissues and pericancerous tissues was 35. 7%and 64. 3%,respectively( P < 0. 01). The expression of DCLK1 mRNA and protein in oral squamous cell carcinoma was higher than that in pericancerous tissues( both P < 0. 01). The DCLK1 gene promoter methylation was negatively correlated with the expressions of DCLK1 mRNA and protein in oral squamous cell carcinoma( r =-0. 767,P < 0. 01; r =-0. 671,P < 0. 01). The methylation status of DCLK1 gene promoter was significantly associated with TNM stage,histological differentiation,and lymph node metastasis in patients with oral squamous cell carcinoma( all P < 0. 05). The 3-year progression-free survival and overall survival of patients with positive DCLK1 gene promoter methylation were higher than those of negative DCLK1 gene promoter methylation( 63. 0% vs 43. 4%,69. 6% vs 47. 0%,both P < 0. 05). Conclusion The methylation status of DCLK1 gene promoter is related to clinicopathological features of oral squamous cell carcinoma,and can affect the prognosis of patients. DCLK1 demethylation may play a role in the occurrence and development of tumor by being involved in the regulation of DCLK1 mRNA expression and protein secretion.
引文
[1]林文远.骨髓增生异常综合征的基因甲基化与去甲基化药物应用的研究现状[J].实用肿瘤杂志,2013,28(4):360-363.
[2] Sarkar S,Popov VL,O'Connell MR,et al. A novel antibody against cancer stem cell biomarker,DCLK1-S,is potentially useful for assessing colon cancer risk after screening colonoscopy[J]. Lab Invest,2017,97(10):1245-1261.
[3] Kalantari E,Lari M,Roudi R,et al. Lgr5High/DCLK1High phenotype is more common in early stage and intestinal subtypes of gastric carcinomas[J]. Cancer Biomark,2017,20(4):563-573.
[4] O'Connell MR,Sarkar S,Singh P. Abstract 1940:Short(S)isoform of cancer-stem-cell marker,DCLK1,is critically required to maintain proliferative/tumorigenic potential of colon cancer cells:identification of associated molecular pathways[J]. Cancer Res,2014,74(19):1940.
[5]桑圣刚,胡敏,荣红.基质金属蛋白酶-9基因启动子甲基化在非小细胞肺癌治疗中的作用[J].中华肺部疾病杂志(电子版),2013,6(3):35-38.
[6]陈芬,陈林林.口腔疣状癌、口腔鳞癌与癌基因关系的研究进展[J].实用临床医学,2016,17(1):92-93.
[7] Yang N,Nijhuis ER,Volders HH,et al. Gene promoter methylation patterns throughout the process of cervical carcinogenesis[J]. Cell Oncol,2016,32(1/2):131-143.
[8]周美玲,贺兼斌,向明钧.双肾上腺素样激酶-1与恶性肿瘤关系的研究进展[J].中国继续医学教育,2017,9(32):84-86.
[9] Gao T,Wang M,Xu L,et al. DCLK1 is up-regulated and associated with metastasis and prognosis in colorectal cancer[J]. J Cancer Res Clin Oncol,2016,142(10):1-10.
[10] Bailey JM,Janivette A,Rasheed ZA,et al. DCLK1marks a morphologically distinct subpopulation of cells with stem cell properties in preinvasive pancreatic cancer[J]. Gastroenterology,2014,146(1):245-256.
[11]孟庆彬,于健春,康维明,等. Doublecortin样激酶-1在胃癌组织中的表达及其与预后关系[J].中国医学科学院学报,2013,35(6):639-644.
[12]王雪玮. DNA甲基化在肿瘤中的调控机制及其在肺癌中的研究进展[J].中国肿瘤生物治疗杂志,2017,24(5):558-566.
[13]赖伊杰,康亚妮.肿瘤细胞DNA甲基化检测技术与研究方法新进展[J].生命科学,2016,28(4):513-520.
[14] Powrózek T,Krawczyk P,Nicos'M,et al. Methylation of the DCLK1 promoter region in circulating free DNA and its prognostic value in lung cancer patients[J]. Clin Transl Oncol,2016,18(4):1-7.
[15] Sarkar S,O'Connell MR,Okugawa Y,et al. FOXD3 regulates CSC marker,DCLK1-S,and invasive potential:prognostic implications in colon cancer[J]. Mol Cancer Res,2017,15(12):1678-1691.
[2] Sarkar S,Popov VL,O'Connell MR,et al. A novel antibody against cancer stem cell biomarker,DCLK1-S,is potentially useful for assessing colon cancer risk after screening colonoscopy[J]. Lab Invest,2017,97(10):1245-1261.
[3] Kalantari E,Lari M,Roudi R,et al. Lgr5High/DCLK1High phenotype is more common in early stage and intestinal subtypes of gastric carcinomas[J]. Cancer Biomark,2017,20(4):563-573.
[4] O'Connell MR,Sarkar S,Singh P. Abstract 1940:Short(S)isoform of cancer-stem-cell marker,DCLK1,is critically required to maintain proliferative/tumorigenic potential of colon cancer cells:identification of associated molecular pathways[J]. Cancer Res,2014,74(19):1940.
[5]桑圣刚,胡敏,荣红.基质金属蛋白酶-9基因启动子甲基化在非小细胞肺癌治疗中的作用[J].中华肺部疾病杂志(电子版),2013,6(3):35-38.
[6]陈芬,陈林林.口腔疣状癌、口腔鳞癌与癌基因关系的研究进展[J].实用临床医学,2016,17(1):92-93.
[7] Yang N,Nijhuis ER,Volders HH,et al. Gene promoter methylation patterns throughout the process of cervical carcinogenesis[J]. Cell Oncol,2016,32(1/2):131-143.
[8]周美玲,贺兼斌,向明钧.双肾上腺素样激酶-1与恶性肿瘤关系的研究进展[J].中国继续医学教育,2017,9(32):84-86.
[9] Gao T,Wang M,Xu L,et al. DCLK1 is up-regulated and associated with metastasis and prognosis in colorectal cancer[J]. J Cancer Res Clin Oncol,2016,142(10):1-10.
[10] Bailey JM,Janivette A,Rasheed ZA,et al. DCLK1marks a morphologically distinct subpopulation of cells with stem cell properties in preinvasive pancreatic cancer[J]. Gastroenterology,2014,146(1):245-256.
[11]孟庆彬,于健春,康维明,等. Doublecortin样激酶-1在胃癌组织中的表达及其与预后关系[J].中国医学科学院学报,2013,35(6):639-644.
[12]王雪玮. DNA甲基化在肿瘤中的调控机制及其在肺癌中的研究进展[J].中国肿瘤生物治疗杂志,2017,24(5):558-566.
[13]赖伊杰,康亚妮.肿瘤细胞DNA甲基化检测技术与研究方法新进展[J].生命科学,2016,28(4):513-520.
[14] Powrózek T,Krawczyk P,Nicos'M,et al. Methylation of the DCLK1 promoter region in circulating free DNA and its prognostic value in lung cancer patients[J]. Clin Transl Oncol,2016,18(4):1-7.
[15] Sarkar S,O'Connell MR,Okugawa Y,et al. FOXD3 regulates CSC marker,DCLK1-S,and invasive potential:prognostic implications in colon cancer[J]. Mol Cancer Res,2017,15(12):1678-1691.