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目的以MCL1基因为例,研究不同亚型在结肠癌组织中差异表达的临床意义。方法采用RT-PCR和WB分别检测正常结肠组织、结肠腺瘤、结肠腺癌组织以及对应的癌旁组织各20例中MCL1基因3种mRNA剪接异构体和3种蛋白亚型的表达水平,进而通过χ2检验分析了这些亚型表达差异在213例肿瘤临床病例中与肿瘤病理特征之间的相关性。结果 MCL1基因亚型1在结肠肿瘤组织及对应旁组织中的表达均高于正常组(mRNA,P=0.009;蛋白,P=0.022),且腺癌组中的表达亦高于腺瘤组(mRNA,P=0.004;蛋白,P=0.034)。亚型2在结肠肿瘤组织及对应旁组织中的表达均高于正常组(mRNA,P=0.002;蛋白,P=0.011),且腺癌组中的表达亦高于腺瘤组(mRNA,P<0.001;蛋白,P=0.007)。亚型3在结肠肿瘤组织及对应旁组织中的表达均高于正常组(mRNA,P=0.027;蛋白,P=0.045),但肿瘤组与对应旁组织之间以及腺癌组与腺瘤组之间的比较均无统计学意义。此外MCL1亚型1和亚型2对结肠癌肿瘤恶性程度呈正相关,而亚型3可以作为肿瘤早期诊断的标志物,但其表达水平与恶性程度无相关性。结论在临床评估结肠癌恶性程度过程中,位于核表达的MCL1亚型2可以作为有效的诊断指标,本研究为MCL1引物设计以及抗体选择提供一定参考价值。
Objective To investigate the clinical significance of differential expression of different subtypes in colon cancer using MCL1 gene as an example. Methods RT-PCR and WB were used to detect the expression levels of three kinds of mRNA splice isoforms and three protein subtypes of MCL1 gene in 20 cases of normal colon, colon adenocarcinoma, colon adenocarcinoma and corresponding paracancerous tissues respectively. Furthermore, the correlation between these subtypes expression and tumor pathological features was analyzed in χ2 test in 213 clinical cases of tumors. Results The expression of MCL1 gene subtype 1 in colorectal tumor tissues and corresponding paracancerous tissues was significantly higher than that in normal tissues (P = 0.009; P = 0.022). The expression of MCL1 in adenocarcinoma was also higher than that in adenoma mRNA, P = 0.004; protein, P = 0.034). The expression of subtype 2 in colon tumor tissue and corresponding paracancerous tissue was higher than that in normal group (P = 0.002; P = 0.011), and the expression in adenocarcinoma was also higher than that in adenoma (P <0.001; protein, P = 0.007). The expression of subtype 3 in colorectal tumor tissues and its adjacent tissues was higher than that in normal tissues (mRNA, P = 0.027; protein, P = 0.045) There was no statistical significance between the two groups. In addition MCL1 subtype 1 and subtype 2 have a positive correlation with the degree of malignancy of colon cancer, while subtype 3 can be used as a marker of early diagnosis of cancer, but its expression level has no correlation with the degree of malignancy. Conclusion In the clinical evaluation of malignant degree of colorectal cancer, MCL1 subtype 2 located in the nucleus can be used as an effective diagnostic indicator. This study provides a reference value for the design of MCL1 primer and antibody selection.