Na+-K+-ATP 酶表达对结直肠癌细胞增殖及侵袭力的影响
张贵海;文坤明;张先平;王轶;胡敏;李少林
【期刊名称】《中国肿瘤临床》 【年(卷),期】2012(039)003
【摘要】目的:探讨Na+-K+-ATP酶α亚单位表达对人结直肠癌细胞增殖及侵袭力的影响,及哇巴因治疗结直肠癌的可能机制.方法:以SW480和SW620细胞为研究对象,予生理浓度的哇巴因(1 nM)干预,应用MTT法、Transwell侵袭小室、生化酶学、实时定量PCR及蛋白免疫印迹等方法检测细胞的增殖及侵袭力,Na+-K+-ATP酶活性及其亚单位(α1、α2及α3)表达水平.结果:与SW480细胞比较,SW620细胞增殖及侵袭力明显增加(P 均<0.05),Na+-K+-ATP酶活性下降,Na+-K+-ATP酶α1亚单位mRNA表达上调(P 均<0.001);而SW480细胞Na+-K+-ATP酶α3亚单位mRNA表达上调(P<0.001);且均与蛋白表达水平一致;哇巴因在24、48及72 h均能显著抑制两种细胞的增殖活力,以48 h效应最强,减弱其侵袭力,并下调两种细胞Na+-K+-ATP酶α1亚单位蛋白表达,增加SW620细胞Na+-K+-ATP酶活性,而对两种细胞Na+-K+-ATP酶α2和α3亚单位蛋白表达及SW480细胞Na+-K+-ATP酶活性无影响.结论:Na+-K+-ATP酶活性下降部分源于α3亚单位表达下调及α1亚单位表达上调,可能共同参与结直肠癌的转移;哇巴因抑制结直肠癌细胞的增殖及侵袭力的机制,可能部分与下调Na+-K+-ATP酶α1亚单位蛋白表达有关.%Objective: To explore the effect of Na+-K+-ATPase subunit expression on the proliferation and invasion of human colorectal cancer cell lines SW480 and SW620 and to determine the potential molecular mechanisms of ouabain
anticancer therapeutics in human colorectal cancer. Methods: SW620 ( Dukes C phase ) and SW480 ( Dukes B phase ) cells were derived from the same patient and treated with or without a physiological concentration of 1 nM of ouabain. The proliferation of SW620 and SW480 cells was evaluated using an MTT reduction assay ( I.e., 3-[ 4, 5 ] -dimethylthiahiazo [ -z-yl ] -3, 5-di- phenytetrazoliumromide ). The invasion capacity was detected using a Transwell chamber. The Nα+-K+-ATPase activity was measured using biochemistry and enzymology. The expression of the Nμ+-K+-ATPase μi-, αi-, and α3-subunits were determined using real-time quantitative PCR and Western blot analysis. Results: The proliferation and invasion of SW620 cells significantly increased compared with those of SW480 cells (P < 0.05 to P < 0.01 ). The enzymatic activity of Na+-K+-ATPase remarkably decreased in SW620 cells ( P < 0.001 ). Furthermore, in SW620 cells, the mRNA-level expression
of
Nμ+-K+-ATPase
a3-subunits
was
significantly
downregulated compared with that of the (Xi- and α2-subunits. However, the Nμ+-K+-ATPase al-subunit expression was significantly upregulated ( P < 0.01 ). Interestingly, the Na+-K+-ATPase α3-sub-unit expression in SW480 cells was upregulated (P < 0.001). Nevertheless, the Nα+, K+-ATPase a2-subunit expressions in the SW480 and SW620 cells were not significantly different ( P > 0.05 ). In addition, the protein expression of the Nα+ -K+-ATPase α1-,α2-, and a 3-subunits were in
line with the accompanying mRNA expression. A physiological concentration of 1 nM of ouabain, a Na+, K+-ATPase inhibitor, could significantly downregulate the protein-level expression of Nα+-K+-ATPase α1-subunits in SW620 and SW480 cells within 48 h, thus inhibiting the growth and decreasing the invasion capacity of SW620 and SW480 cells. Ouabain could increase Nα+-K+-ATPase activity in SW620 cells. The Nα+-K+-ATPase α2- and α3-subunit expression in the SW620 and SW480 cells treated with ouabain for 48 h were not significantly different. Conclusion: The aberrant activity of Nα+-K+-ATPase might involve tumor metastasis in human colorectal cancer, which is partly ascribed to the abnormal expression of the Nα+-K+-ATPase α1- and α3-subunits. However, ouabain could effectively decrease the proliferation and invasion of SW480 and SW620 cells by downregulating the Nα+-K+-ATPase a 1-subunits. 【总页数】5页(121-125)
【关键词】结直肠癌;腺苷三磷酸酶;哇巴因;增殖;侵袭 【作者】张贵海;文坤明;张先平;王轶;胡敏;李少林
【作者单位】重庆医科大学基础医学院放射医学教研室;重庆医科大学附属第一医院胃肠外科;遵义医学院组胚教研室;重庆医科大学基础医学院放射医学教研室;重庆医科大学基础医学院放射医学教研室;重庆医科大学基础医学院放射医学教研室
【正文语种】中文
【中图分类】 【文献来源】
https://www.zhangqiaokeyan.com/academic-journal-cn_chinese-journal-clinical-oncology_thesis/0201236806128.html 【相关文献】
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Na+-K+-ATP 酶表达对结直肠癌细胞增殖及侵袭力的影响
