CAJ | 학술논문

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（Westernblot）、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Westernblot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Westernblot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。
목적：검측불동분화정도결장암조직중수통도단백（AQP）‐5여내약인자적표체급상호관계。방법대45례결장암조직、36례암방조직채용실시형광정량PCR（RT‐PCR）、단백면역인적법（Westernblot）、면역조직화학염색검측AQP‐5급내약인자P‐당단백（P‐gp）、곡광감태전이매（GST‐π）、탁복이구매（TopoⅡ）、흉감산적한속매（TS）적표체정황。결과면역조직화학염색결과표명，AQP‐5주요분포우세포막화세포질；RT‐PCR화Westernblot검측결과현시，결장암조직중AQP‐5적표체현저고우암방대조조（P＜0．05），차표체량수분화정도강저이승고（P＜0．05）。면역조직화학염색발현，P‐gp주요분포우세포막화세포질，GST‐π주요분포우세포핵화세포질，TopoⅡ주요분포우세포핵，TS주요분포우세포질。RT‐PCR화Westernblot검측결과현시，결장암조직중내약인자표체량현저고우암방조직（P＜0．05），기중P‐gp、GST‐π、TopoⅡ수착분화정도강저표체증가，TS재불동분화정도결장암조직중표체변화불명현（P＞0．05）。AQP‐5여P‐gp、GST‐π、TopoⅡ적표체정정상관（P＜0．05），여TS표체무상관성（P＞0．05）。결론AQP‐5여내약인자P‐gp、GST‐π、TopoⅡ、TS단백재결장암조직중고도표체，가능촉진료결장암적전이화진전。
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 aqua‐porin 5 and resistance genes P‐gp ,GST‐π,TopoⅡ ,and TS in human 45 cases colon cancer tissues with different differentiation de‐gree and 36 cases of adjacent mucosa tissues as well as 58 cases of normal colonic epithelium were detected by quantitative RT‐PCR ,Western blot and immunohistochemistry .Results Immunohistochemistry results showed that AQP‐5 distributed mainly in the cell membrane and the cytoplasm .Fluorescence quantitative RT‐PCR and Western blot showed that AQP‐5 expression could not be detected in adjacent mucosa tissues and normal colonic epithelium tissues .The AQP‐5 expression 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 P‐gp distributed mainly in the cell membrane and the cytoplasm ,GST‐πmainly distributed in the nuclei and cytoplasm ,Topo Ⅱ main‐ly distributed in the nucleus ,and TS distributed mainly in the cytoplasm .Fluorescence quantitative RT‐PCR and Western blot re‐sults 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 Topo Ⅱ expression were negative‐ly correlated with the differentiation degree of colon cancer tissues ,the more poor differentiation level of tissue ,the higher expres‐sion level of P‐gp ,GST‐π ,Topo Ⅱ .However ,the expression level of TS did not change significantly in different differentiation de‐gree colon cancer tissues (P>0 .05) .Positive correlation was found between the expression of AQP‐5 and P‐gp ,GST‐π,Topo Ⅱ(P<0 .05) .Negative correlation was found between the expression of AQP‐5 and TS (P>0 .05) .Conclusion The AQP‐5 and re‐sistance gene expression were increased in colon cancer tissues .The AQP‐5 expression level was higher in colon cancer compared with adjacent control or normal tissues ,which may promote the transfer and progress of colon cancer .