摘要
目的 探讨食管癌组织中乙酰基转移酶2A(KAT2A)表达与Survivin蛋白乙酰化水平,分析二者在食管癌发生过程中的相关性。方法 选取70例食管鳞癌组织及其癌旁正常食管黏膜组织。实时荧光定量聚合酶链反应(qRT-PCR)检测Survivin和KAT2A mRNA的表达,免疫组化及Western blotting检测Survivin和KAT2A的蛋白表达,免疫荧光验证Survivin和KAT2A在癌组织中的定位及表达;免疫共沉淀检测Survivin乙酰化。分析Survivin蛋白乙酰化与KAT2A的相关性及其二者与食管癌临床病理特征的关系。构建KAT2A RNA干扰序列,脂质体介导法转染至食管癌EC109细胞。应用qRT-PCR和Western blotting检测转染siRNA-KAT2A的食管癌EC109细胞中KAT2A mRNA和蛋白表达水平,免疫共沉淀检测EC109细胞中Survivin蛋白乙酰化水平。结果 食管癌组织中Survivin和KAT2A mRNA相对表达量高于癌旁组织(P<0.05);食管癌组织中Survivin和KAT2A蛋白呈高表达,而癌旁组织则呈低表达。免疫荧光证实Survivin和KAT2A蛋白在癌组织中同时呈现高表达状态,二者主要表达于细胞质中,部分表达于细胞核中。食管癌组织中Survivin蛋白发生了乙酰化,且食管癌组织中的乙酰化率显著高于癌旁组织(P<0.05);在食管癌组织中Survivin乙酰化和KAT2A表达呈正相关(r=0.517,P<0.05);Survivin乙酰化和KAT2A表达与食管癌的TNM分期、分化程度和淋巴结转移有关(P<0.05)。体外实验显示,RNA干扰下调KAT2A对食管癌细胞Survivin mRNA和蛋白表达量均无影响(P>0.05),但Survivin蛋白的乙酰化水平显著降低(P<0.05)。结论 在食管病变过程中KAT2A参与了Survivin乙酰化修饰,其高表达可能是Survivin乙酰化修饰重要的前期分子事件,在食管癌的诊治中具有潜在应用价值。
Objective To investigate the expression of acetyltransferase 2A(KAT2A) and survivin protein acetylation levels in esophageal cancer tissue, and to analyze the correlation and clinical significance during the onset of esophageal cancer. Methods Seventy cases of esophageal squamous carcinoma tissues and adjacent cancer tissues were collected. The survivin and KAT2A mRNA expression was determined by reverse transcription and real-time PCR(qRT-PCR), the expression levels of survivin and KAT2A protein were detected by Immunohistochemistry and Western blotting. Fluorescence immunohistochemistry(IF) verified the location and expression of survivin and KAT2A in cancerous tissues.The survivin protein acetylation was detected by co-immunoprecipitation. The correlation between survivin protein acetylation and KAT2A, and the relationship between them and the clinicopathological characteristics of esophageal cancer were analyzed. KAT2A RNA interference sequences was constructed, then transfected into EC109 cells using liposome-mediated methods. qRT-PCR and Westrern blotting were used to detect the expression level of KAT2A mRNA and protein in esophageal cancer EC109 cells transfected with siRNA-KAT2A. The acetylation level of survivin protein was detected by co-immunoprecipitation. Results The relative expression of survivin and KAT2A mRNA was higher in esophageal cancer tissues than that in adjacent cancer tissues(P<0.05). Survivin and KAT2A protein were highly expressed in esophageal cancer tissues, whereas low expressed in adjacent cancer tissues.The survivin protein was acetylated in esophageal cancer tissues. Immunofluorescence confirmed that survivin and KAT2A protein both show a high expression state in cancer tissues, which are mainly expressed in the cytoplasm, partially expressed in the nucleus, and the acetylation rate was significantly higher than that in adjacent cancer tissues(P<0.05). Survivin acetylation and KAT2A were related in esophageal cancer tissues, and it was positive correlation(r=0.517,P<0.05). Survivin protein acetylation and KAT2A are closely related to tumor TNM staging, differentiation grade and lymph node metastasis(P<0.05). The result of vitro experiments showed that there was no change in survivin mRNA and protein expression after down-regulation of KAT2A by RNA interference(P>0.05), but the acetylation level of survivin protein was significantly reduced(P<0.05). Conclusion Acetyltransferase KAT2A is involved in the survivin acetylation modification during the esophageal lesion. High expression of KAT2A may be an important molecular event of survivin acetylation modification and has potential value in the diagnosis and treatment of esophageal cancer.
作者
梁宗英
赵宝山
郑竞雄
侯继申
孙光蕊
LIANG Zongying;ZHAO Baoshan;ZHENG Jingxiong;HOU Jishen;SUN Guangrui(Department of Thoracic Surgery,Affiliated Hospital of Chengde Medical College,Chengde 067000,China)
出处
《临床肿瘤学杂志》
CAS
2023年第1期16-22,共7页
Chinese Clinical Oncology
基金
承德市科学技术研究与发展计划项目(201606A036)
河北省医学科学研究重点课题计划项目(20210985)。
