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华丛书, 马冬春, 陈海等. p16与非小细胞肺癌临床病理的关系及5-Aza-CdR对肺癌A549细胞中p16表达的影响[J]. 四川大学学报(医学版), 2018, 49(3): 404-407.
引用本文: 华丛书, 马冬春, 陈海等. p16与非小细胞肺癌临床病理的关系及5-Aza-CdR对肺癌A549细胞中p16表达的影响[J]. 四川大学学报(医学版), 2018, 49(3): 404-407.
HUA Cong-shu, MA Dong-chun, CHEN Hai. et al. The Relationship of Clinicopathology of p16 in Non-small Cell Lung Cancer and 5-2′-deoxy cytidine-nitrogen Impurity on the Influence of p16 Expression in Lung Cancer A549 Cells[J]. Journal of Sichuan University (Medical Sciences), 2018, 49(3): 404-407.
Citation: HUA Cong-shu, MA Dong-chun, CHEN Hai. et al. The Relationship of Clinicopathology of p16 in Non-small Cell Lung Cancer and 5-2′-deoxy cytidine-nitrogen Impurity on the Influence of p16 Expression in Lung Cancer A549 Cells[J]. Journal of Sichuan University (Medical Sciences), 2018, 49(3): 404-407.

p16与非小细胞肺癌临床病理的关系及5-Aza-CdR对肺癌A549细胞中p16表达的影响

The Relationship of Clinicopathology of p16 in Non-small Cell Lung Cancer and 5-2′-deoxy cytidine-nitrogen Impurity on the Influence of p16 Expression in Lung Cancer A549 Cells

  • 摘要: 目的 探讨抑癌基因p16在非小细胞肺癌(non-small cell lungcancer,NSCLC)组织中的表达与临床病理参数间的联系,进一步研究DNA甲基转移酶抑制剂5-氮杂-2′-脱氧胞苷(5-Aza-2′-deoxycytidine,5-Aza-CdR)在人肺癌A549细胞中对p16 表达的影响。方法 采用免疫组化SP法检测76例NSCLC组织及其癌旁正常组织中p16 蛋白的表达情况,并分析不同病理参数癌组织中p16蛋白表达的差异;采用MSP法检测不同浓度(0~16 μmol/L)5-Aza-CdR处理后的A549细胞中P16基因启动子区DNA的甲基化状态,并采用Western blot 法检测5-Aza-CdR对A549细胞中p16 表达的影响。结果 76例NSCLC组织中,32例(42.11%)p16 蛋白表达阳性,低于癌旁正常组织(59例,77.63%)中阳性表达(P<0.05);不同病理组织分级、肿瘤分化程度、临床TNM分期及有无淋巴结转移的NSCLC组织中p16 的阳性表达率差异有统计学意义(P<0.05);在A549细胞中p16蛋白表达和非甲基化产物均呈低表达状态,5-Aza-CdR作用后,p16蛋白和其非甲基化产物表达均上调,且随着5-Aza-CdR浓度的升高呈增多趋势。结论 p16 在鳞癌、低分化、有淋巴结转移及Ⅲ~Ⅳ期NSCLC组织中低表达,提示其失活可能与NSCLC的进一步进展有关,5-Aza-CdR可诱导A549细胞中p16蛋白和非甲基化产物表达增加。

     

    Abstract: Objective To investigate the relationship between expression of tumor suppressor gene p16 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. p16 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 (P<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.

     

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