食管癌中KIAA1522基因的转录本及蛋白表达分析

Analysis of expression of KIAA1522 in esophageal squamous cell carcinoma

目的:检测食管鳞癌组织及细胞系中KIAA1522基因不同转录本的表达及丰度,确定其蛋白的表观分子量,为进一步研究KIAA1522在食管癌中的表达改变及功能提供实验依据。方法:使用反转录PCR(RT-PCR)及实时荧光定量PCR(qPCR)检测食管癌组织及细胞系中KIAA1522基因4个不同转录本的表达及丰度;以特异性si RNA敲降食管癌细胞中KIAA1522的表达,并在模式细胞中过表达KIAA1522,通过Western blot检测确定KIAA1522的表观分子量,并通过质谱分析验证KIAA1522条带的序列组成。结果:KIAA1522的转录本T1和T4在食管鳞癌组织及多个细胞系中均有表达,其中T1的表达丰度相对最高。Western blot检测中观察到的分子量为170 k Da的条带为KIAA1522基因特异性的蛋白表达产物,同时质谱分析结果也证实该条带为KIAA1522多肽。结论:T1为食管鳞癌组织及细胞系中KIAA1522基因的优势转录本,KIAA1522蛋白的表观分子量为170 kDa。

OBJECTIVE:To detect the expression and abundance of different transcripts of KIAA1522gene in esophageal squamous cell carcinoma(ESCC)tissues and cell lines,and to determine the apparent molecular weight of the protein.METHODS:The expression and abundance of four different transcripts of KIAA1522gene in ESCC tissues and cell lines were detected by reverse transcription PCR(RT-PCR)and quantitative real-time PCR(qPCR).Molecular weight of the KIAA1522protein was determined by Western blot together with experiments that involved KIAA1522gene specificknockdown or exogenous over-expression.The sequence of KIAA1522protein was confirmed by mass spectrometry.RESULTS:Both T1and T4transcripts of KIAA1522were expressed in ESCC tissues and cell lines,but T1was the mostabundant.The170kDa band observed in the Western blot was the KIAA1522gene-specific protein expression product.CONCLUSION:T1was the predominant transcript of KIAA1522gene in ESCC tissues and cell lines.The apparent molecular weight of KIAA1522protein was170kDa.