赖氨酰氧化酶体内外对乳腺癌增殖和侵袭作用的研究  

Effect of Lysyl Oxidase on Human Breast Cancer Proliferation and Invasion in vivo and in vitro

作　　者：杨华伟[1] 刘剑仑[1] 李景涛[1] 韦薇[1] 唐玮[1] 蒋奕[1] 

机构地区：[1]广西医科大学附属肿瘤医院乳腺中心,南宁530021

出　　处：《医学研究杂志》2013年第4期73-77,共5页Journal of Medical Research

基　　金：广西科学研究与技术开发计划基金资助项目(0816004-14)

摘　　要：目的探讨赖氨酰氧化酶(lysyl oxidase,LOX)对人乳腺癌细胞的生长增殖、侵袭及转移能力的影响及可能作用的机制。方法构建LOX基因的特异性慢病毒干扰载体(LOX-RNAi-LV),稳定转染至乳腺癌细胞MDA-MB-231,实验设3个组,即空白组(Con)、干扰组(RNAi)、阴性对照组(Mock),四甲基偶氮唑蓝(MTT)法、细胞集落形成实验及流式细胞仪观察细胞生长及增殖情况,细胞侵袭迁移实验(Transwell)检测细胞侵袭及迁徙能力改变;免疫组织化学法检测111例人乳腺癌组织、癌旁乳腺组织及20例乳腺良性病变组织中LOX、基质金属蛋白酶(MMP-2,MMP-9)的表达,并对LOX与临床病理资料及MMP-2、MMP-9的相关性进行分析。结果转染72h后,MTT法显示干扰组细胞生长明显受抑制;细胞集落形成实验显示干扰组的细胞集落形成率明显低于空白组和对照组;Transwell侵袭和迁移实验结果显示RNAi组穿过Transwell小室滤过膜细胞数分别为47±2和63±2,差异有统计学意义(P=0.000);流式细胞仪测定细胞周期,3组之间差异无统计学意义。LOX蛋白在人乳腺癌、癌旁乳腺组织及乳腺良性肿瘤组织中的表达率分别为48.64%(54/111)、26.13%(29/111)、20.00%(4/20),乳腺癌组织中LOX蛋白的表达率明显高于癌旁乳腺组织及乳腺良性肿瘤组织(P=0.019)。LOX蛋白在不同肿瘤大小、临床分期、淋巴结转移的表达率存在显著差异。相关分析显示,LOX蛋白表达与MMP-2(r=0.262,P=0.005)、MMP-9(r=0.424,P=0.000)蛋白之间呈显著正相关。结论 LOX可以促进乳腺癌的侵袭转移;LOX和MMP-2、MMP-9转移因子可能具有协同促进作用,促进了乳腺癌的侵袭转移。Objective To investigate the effect of lysyl oxidase （LOX） on human breast cancer cell growth, proliferation, invasion and metastasis and to explore its possible role in the mechanism of breast cancer. Methods Lysyl oxidase gene - specific lentiviral RNA interference vetor（ LOX - RNAi - LV） was designed and synthesized, which was stably transfected into breast cancer cell lines MDA - MB -231. The breast cancer cells were divided into three groups：the experimental group （RNAi）, the negative control group （Mock） and the blank control group （Con）. The changes of cell growth and proliferation of breast cancer cells MDA - MB - 231 were obsered by MTT assay, cell colony formation and Flow cytometric. The cells migration and invasion abilities were measured by cell migration and invasion test（Transwell）. 111 cases of human breast cancer tissue and cancer- adjacent breast tissues and 20 cases of benign lesion tissues of LOX, MMP -2 and MMP -9 were detected by immunohistochemistry, and the relationship of LOX between clinicopathological character- istics and MMP -2, MMP -9 was analyzed. Results After transfection 48h, the cell growth inhibition rates of RNAi group was signifi- cantly higher than that of the Mock group and the Con group （ P = 0. 000）. Colony formation experiments showed that RNAi group cell col- ony formation rate was significantly lower than the Con group and the Mock group. Transwell results showed that RNAi group through the Transwell chamber filtration membrane cells were 47 ＋ 2 and 63 ＋ 2, and the difference was significant （ P = 0. 000）. Cell cycle was ana- lyzed by flow cytometry, which showed there was no difference among the three groups. The expression of LOX protein in breast cancer, cancer - adjacent breast tissues and benign breast tumor were 48.64% （54/111） , 26.13% （29/111） , 20.00% （4/20）. The expres- sion of LOX protein in breast cancer was significantly higher than that in cancer - adjacent breast tissues and benign lesion tissues （ P =

关 键 词：乳腺肿瘤 赖氨酰氧化酶 增殖及侵袭 

分 类 号：R737.9[医药卫生—肿瘤]