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[非小细胞肺癌中p16的临床病理关系及5-氮杂-2'-脱氧胞苷氮杂质对肺癌A549细胞p16表达的影响]

[The Relationship of Clinicopathology of p16 in Non-small Cell Lung Cancer and 5-nitrogen impurity-2'-deoxycytidine-nitrogen Impurity on the Influence of p16 Expression in Lung Cancer A549 Cells].

作者信息

Hua Cong-Shu, Ma Dong-Chun, Chen Hai, Zhang Chao-Dong, Wang Wen-Sheng, Ding Xiang, Zhao Jian, Wu Feng

机构信息

Department of Thoracic Surgery,Anhui Province Chest Hospital,Hefei 230022,China.

出版信息

Sichuan Da Xue Xue Bao Yi Xue Ban. 2018 May;49(3):404-407.

Abstract

OBJECTIVE

To investigate the relationship between expression of tumor suppressor gene 16 in non-small cell lung cancer (NSCLC) tissues and clinicopathological parameters,to further study on DNA methyltransferase inhibitors 5-nitrogen impurity-2'-deoxycytidine (5-Aza-CdR) in human lung cancer cell line A549 in regulating the expression of p16.

METHODS

The expression of p16 protein in 76 cases of NSCLC tissues and normal tissue adjacent to carcinoma were detect by immunohistochemical SP method and the differences of p16 protein expression were analyzed. 16 gene promoter region of DNA methylation status were detect by MSP method in 5-Aza-CdR processing A549 cells,the expression of p16 in A549 lung cancer cell and effect of 5-Aza-CdR were detect by Western blot method.

RESULTS

32 cases (42.11%) of p16 protein expression was positive,significantly lower than that of the normal tissue adjacent to carcinoma (positive expression in 59 cases,77.63%) in 76 cases of NSCLC tissues; There were statistically significant differences (<0.05) in the positive expression rates of p16 in NSCLC tissues with different pathological tissue grading,tumor differentiation degree,clinical TNM stage and lymph node metastasis. In A549 cells,p16 protein expression and non-methylated products were both in low expression states. After treated with 5-Aza-CdR,the expression of p16 protein and its non-methylated products were up-regulated,with the increase of 5-Aza-CdR concentration.

CONCLUSION

The low expression of p16 in NSCLC tissues with squamous cell carcinomas,low differentiation,lymph node metastasis and phase Ⅲ-Ⅳ,which may prompt the deactivation and cause further progression of NSCLC,5-Aza-CdR could induce the expression of p16 protein and non-methylated products in A549 cells.

摘要

目的

探讨抑癌基因16在非小细胞肺癌(NSCLC)组织中的表达与临床病理参数的关系,进一步研究DNA甲基转移酶抑制剂5-氮杂-2'-脱氧胞苷(5-Aza-CdR)对人肺癌细胞系A549中p16表达的调控作用。

方法

采用免疫组织化学SP法检测76例NSCLC组织及癌旁正常组织中p16蛋白的表达,并分析p16蛋白表达的差异。采用甲基化特异性PCR(MSP)法检测5-Aza-CdR处理A549细胞后16基因启动子区域的DNA甲基化状态,采用蛋白质免疫印迹法(Western blot)检测A549肺癌细胞中p16的表达及5-Aza-CdR的作用效果。

结果

76例NSCLC组织中,32例(42.11%)p16蛋白表达为阳性,显著低于癌旁正常组织(59例阳性表达,77.63%);不同病理组织分级、肿瘤分化程度、临床TNM分期及淋巴结转移的NSCLC组织中p16阳性表达率差异有统计学意义(<0.05)。在A549细胞中,p16蛋白表达及非甲基化产物均处于低表达状态。经5-Aza-CdR处理后,p16蛋白及其非甲基化产物的表达上调,且随5-Aza-CdR浓度增加而增加。

结论

p16在肺鳞癌、低分化、有淋巴结转移及Ⅲ-Ⅳ期NSCLC组织中低表达,可能促使NSCLC失活并进一步进展,5-Aza-CdR可诱导A549细胞中p16蛋白及其非甲基化产物的表达。

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