摘要
目的检测不同分化程度结肠癌组织中水通道蛋白(AQP)-5与耐药因子的表达及相互关系。方法对45例结肠癌组织、36例癌旁组织采用实时荧光定量PCR(RT-PCR)、蛋白免疫印迹法(Western blot)、免疫组织化学染色检测AQP-5及耐药因子P-糖蛋白(P-gp)、谷胱甘肽转移酶(GST-π)、拓扑异构酶(TopoⅡ)、胸苷酸的限速酶(TS)的表达情况。结果免疫组织化学染色结果表明,AQP-5主要分布于细胞膜和细胞质;RT-PCR和Western blot检测结果显示,结肠癌组织中AQP-5的表达显著高于癌旁对照组(P<0.05),且表达量随分化程度降低而升高(P<0.05)。免疫组织化学染色发现,P-gp主要分布于细胞膜和细胞质,GST-π主要分布于细胞核和细胞质,TopoⅡ主要分布于细胞核,TS主要分布于细胞质。RT-PCR和Western blot检测结果显示,结肠癌组织中耐药因子表达量显著高于癌旁组织(P<0.05),其中P-gp、GST-π、TopoⅡ随着分化程度降低表达增加,TS在不同分化程度结肠癌组织中表达变化不明显(P>0.05)。AQP-5与P-gp、GST-π、TopoⅡ的表达呈正相关(P<0.05),与TS表达无相关性(P>0.05)。结论 AQP-5与耐药因子P-gp、GST-π、TopoⅡ、TS蛋白在结肠癌组织中高度表达,可能促进了结肠癌的转移和进展。
Objective To investigate the expression patterns and significance of aquaporin 5（AQP-5）and multidrug-resistance associated genes in human colon cancer with different differentiation degree and their correlation.Methods The expression of aquaporin 5and resistance genes P-gp,GST-π,TopoⅡ,and TS in human 45 cases colon cancer tissues with different differentiation degree and 36 cases of adjacent mucosa tissues as well as 58 cases of normal colonic epithelium were detected by quantitative RTPCR,Western blot and immunohistochemistry.Results Immunohistochemistry results showed that AQP-5distributed mainly in the cell membrane and the cytoplasm.Fluorescence quantitative RT-PCR and Western blot showed that AQP-5expression could not be detected in adjacent mucosa tissues and normal colonic epithelium tissues.The AQP-5expression level was higher in colon cancer tissues compared with adjacent mucosa tissues and normal colonic epithelium tissues（P〈0.05）,and the expression intensity was correlated with the differentiation degree of colon cancer tissues（P〈0.05）.The results of immunohistochemistry indicated that Pgp distributed mainly in the cell membrane and the cytoplasm,GST-πmainly distributed in the nuclei and cytoplasm,TopoⅡ mainly distributed in the nucleus,and TS distributed mainly in the cytoplasm.Fluorescence quantitative RT-PCR and Western blot results showed that the expression levels of all resistance genes detected were higher in colon cancer tissues compared with adjacent mucosa tissues and normal colonic epithelium tissues（P〈0.05）.Furthermore,P-gp,GST-π,and TopoⅡ expression were negatively correlated with the differentiation degree of colon cancer tissues,the more poor differentiation level of tissue,the higher expression level of P-gp,GST-π,TopoⅡ.However,the expression level of TS did not change significantly in different differentiation degree colon cancer tissues（P〉0.05）.Positive correlation was found between the expression of AQP-5and P-gp,GST-π,Topo Ⅱ（P〈0.05）.Negative correlation was found between the expression of AQP-5and TS（P〉0.05）.Conclusion The AQP-5and resistance gene expression were increased in colon cancer tissues.The AQP-5expression level was higher in colon cancer compared with adjacent control or normal tissues,which may promote the transfer and progress of colon cancer.
出处
《重庆医学》
CAS
北大核心
2015年第5期644-646,共3页
Chongqing medicine
