非小细胞肺癌组织中抑癌基因RUNX3启动子甲基化与肿瘤侵袭相关基因E-Cad、MMP-7和TIMP-1蛋白表达的关系
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摘要
目的:探讨非小细胞肺癌组织中抑癌基因RUNX3启动子甲基化与肿瘤侵袭相关基因E-Cad、MMP-7和TIMP-1蛋白表达的关系。方法:收集并分析我院2017年1月至2018年1月期间收治入院的65例非小细胞肺癌患者组织,采用巢氏甲基化特异性PCR(n MSP)法检测RUNX3启动子甲基化情况;链霉菌素-生物素过氧化物酶(S-P)染色法检测肿瘤侵袭相关基因E-Cad、MMP-7和TIMP-1蛋白表达并计数,并回顾性分析RUNX3启动子甲基化状态与临床病理特征、肿瘤侵袭相关基因表达的关系。结果:65例NSCLC组织样本中有27例出现RUNX3启动子的甲基化(41. 54%)。RUNX3启动子甲基化与病理类型有关,腺癌(81. 48%)高于鳞癌(18. 52%,P=0. 003);且与临床分期有关(P=0. 003),主要分布于Ⅲ期(55. 56%)和Ⅳ期(33. 33%)。S-P染色并计数结果显示,E-Cad、MMP-7和TIMP-1蛋白表达阳性率分别为56. 92%、46. 15%和52. 31%;且在RUNX3基因启动子甲基化组中阳性表达率与非甲基化组中阳性率相比差异均具有统计学意义(P<0. 05)。结论:RUNX3基因启动子的甲基化与NSCLC的侵袭和转移密切相关,有望为NSCLC的临床诊疗提供新的思路。
Objective: To investigate the relationship between the methylation of RUNX3 promoter and the expression of E-Cad,MMP-7 and TIMP-1 in non-small cell lung cancer tissues. Methods: The tissues of 65 patients with non-small cell lung cancer admitted from January 2017 to January 2018 in our hospital were collected and analyzed. The methylation of RUNX3 promoter was detected by nested methylation-specific PCR( n MSP) method. Streptavidin-biotin peroxidase( S-P) staining was used to detect the expression of tumor invasive genes E-Cad,MMP-7 and TIMP-1 and counted. The methylation status of RUNX3 promoter was analyzed retrospectively for the relationship with the pathological characteristics and tumor invasion-related gene expression. Results: The methylation of the RUNX3 promoter occurred in 27 of 65 NSCLC tissue samples( 41. 54%). RUNX3 promoter methylation was associated with histological type,adenocarcinoma( 81. 48%) was higher than squamous cell carcinoma( 18. 52%,P = 0. 003); RUNX3 promoter methylation was also related to clinical stage( P = 0. 003),mainly distributed in stageⅢ( 55. 56%) and Ⅳ period( 33. 33%).S-P staining and counting results showed that the positive rate of E-Cad,MMP-7 and TIMP-1 protein expression were 56. 92%,46. 15%and 52. 31%,respectively; and the positive expression rate in the RUNX3 gene promoter and non-methylation group were statistically different( P<0. 05). Conclusion: The methylation of RUNX3 gene promoter is closely related to the invasion and metastasis of NSCLC,and it is expected to provide new ideas for clinical diagnosis and treatment of NSCLC.
Objective: To investigate the relationship between the methylation of RUNX3 promoter and the expression of E-Cad,MMP-7 and TIMP-1 in non-small cell lung cancer tissues. Methods: The tissues of 65 patients with non-small cell lung cancer admitted from January 2017 to January 2018 in our hospital were collected and analyzed. The methylation of RUNX3 promoter was detected by nested methylation-specific PCR( n MSP) method. Streptavidin-biotin peroxidase( S-P) staining was used to detect the expression of tumor invasive genes E-Cad,MMP-7 and TIMP-1 and counted. The methylation status of RUNX3 promoter was analyzed retrospectively for the relationship with the pathological characteristics and tumor invasion-related gene expression. Results: The methylation of the RUNX3 promoter occurred in 27 of 65 NSCLC tissue samples( 41. 54%). RUNX3 promoter methylation was associated with histological type,adenocarcinoma( 81. 48%) was higher than squamous cell carcinoma( 18. 52%,P = 0. 003); RUNX3 promoter methylation was also related to clinical stage( P = 0. 003),mainly distributed in stageⅢ( 55. 56%) and Ⅳ period( 33. 33%).S-P staining and counting results showed that the positive rate of E-Cad,MMP-7 and TIMP-1 protein expression were 56. 92%,46. 15%and 52. 31%,respectively; and the positive expression rate in the RUNX3 gene promoter and non-methylation group were statistically different( P<0. 05). Conclusion: The methylation of RUNX3 gene promoter is closely related to the invasion and metastasis of NSCLC,and it is expected to provide new ideas for clinical diagnosis and treatment of NSCLC.
引文
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[4] Xiao L,Lan X,Shi X,et al. Cytoplasmic rap1 mediates cisplatin resistance of non-small cell lung cancer[J]. Cell Death Dis,2017,8(5):e2803.
[5] Ma YN,Zhang HY,Fei LR,et al. Satb2 suppresses non-small cell lung cancer invasiveness by g9a[J]. Clin Exp Med,2018,18(1):37-44.
[6] Wang C,Ma W,Wei R,et al. Clinicopathological significance of chfr methylation in non-small cell lung cancer:A systematic review and meta-analysis[J]. Oncotarget,2017,8(65):109732-109739.
[7] Wang XZ,Cheng Y,Wang KL,et al. Peperomin e reactivates silenced tumor suppressor genes in lung cancer cells by inhibition of DNA methyltransferase[J]. Cancer Sci,2016,107(10):1506-1519.
[8] Huang T,Li J,Zhang C,et al. Distinguishing lung adenocarcinoma from lung squamous cell carcinoma by two hypomethylated and three hypermethylated genes:A meta-analysis[J]. PLo S One,2016,11(2):e0149088.
[9] Passaniti A,Brusgard JL,Qiao Y,et al. Roles of runx in hippo pathway signaling[J]. Adv Exp Med Biol,2017,962:435-448.
[10] Shin EJ,Kim HJ,Son MW,et al. Epigenetic inactivation of runx3in colorectal cancer[J]. Ann Surg Treat Res,2018,94(1):19-25.
[11] Liu S,Liu B. Overexpression of nitrogen permease regulator like-2(nprl2)enhances sensitivity to irinotecan(cpt-11)in colon cancer cells by activating the DNA damage checkpoint pathway[J]. Med Sci Monitor,2018,24:1424-1433.
[12]黄冬云,许文景,周锐,等. Cd163分子在非小细胞肺癌患者组织中表达及其对肺癌细胞侵袭和增殖的影响[J].中国免疫学杂志,2016,32(3):410-413.Huang DY,Xu WJ,Zhou R,et al. Expression of Cd163 in nonsmall cell lung cancer tissues and its effect on invasion and proliferation of lung cancer cells[J]. Chin J Immunol,32(3):410-413.
[13]陈巧巧,孙一奎,王峰.联合检测外周血游离lunxmrna、scc和β2-mg对非小细胞肺癌的早期诊断价值[J].中国免疫学杂志,2016,32(5):720-721.Chen QQ,Sun YK,Wang F. Combined detection of free lunxmrna,scc andβ2-mg in peripheral blood for early diagnosis of non-small cell lung cancer[J]. Chin J Immunol,2016,32(5):720-721.
[14] He LY,Zhang H,Wang ZK,et al. Diagnostic and prognostic significance of e-cadherin and ki-67 expression in non-small cell lung cancer patients[J]. Eur Rev Med Pharmacol,2016,20(18):3812-3817.
[15] Luo T,Wang L,Wu P,et al. Downregulated vimentin and upregulated e-cadherin in t1 stage non-small-cell lung cancer:Does it suggest a mesenchymal-epithelial transition?[J].Neoplasma,2017,64(5):693-699.
[16] Blanco-Prieto S,Barcia-Castro L,Paez de la Cadena M,et al.Relevance of matrix metalloproteases in non-small cell lung cancer diagnosis[J]. BMC Cancer,2017,17(1):823.
[17] Ghoshal-Gupta S,Kutiyanawalla A,Lee BR,et al. Timp-1 downregulation modulates mir-125a-5p expression and triggers the apoptotic pathway[J]. Oncotarget,2018,9(10):8941-8956.
[18] Jung YJ,Katilius E,Ostroff RM,et al. Development of a protein biomarker panel to detect non-small-cell lung cancer in korea[J]. Clin Lung Cancer,2017,18(2):e99-e107.
[19] Lee J,Sohn EJ,Yoon SW,et al. Anti-metastatic effect of dehydrocorydaline on h1299 non-small cell lung carcinoma cells via inhibition of matrix metalloproteinases and b cell lymphoma 2[J]. Phytotherapy Res,2017,31(3):441-448.
[20] Winkler K,Ramer R,Dithmer S,et al. Fatty acid amide hydrolase inhibitors confer anti-invasive and antimetastatic effects on lung cancer cells[J]. Oncotarget,2016,7(12):15047-15064.
[21] Chung JH,Ahn CB,Son KH,et al. Simulated microgravity effects on nonsmall cell lung cancer cell proliferation and migration[J].Aerosp Med Hum Performance,2017,88(2):82-89.