摘要
目的:探讨Nm23-H1对乳腺癌细胞上皮-间充质转化(EMT)、侵袭和转移的影响。方法:MDA-MB-231接种于6孔板,采用包装对照和Nm23-H1慢病毒感染,构建对照和Nm23-H1过表达细胞系,分别为对照组和Nm23-H1组。采用细胞计数试剂盒(CCK-8)分析两组细胞增殖能力;采用蛋白质印迹法(Western blot)分析两组细胞上皮型钙黏蛋白(E-cadherin)、波形蛋白(Vimentin)和神经型钙黏蛋白(N-cadherin)蛋白表达水平。采用Transwell分析两组细胞侵袭能力;采用异体移植瘤实验分析两组细胞在体内的转移能力。计量数据比较采用t检验。结果:对照组细胞Nm23-H1表达水平(0.87±0.18)明显低于Nm23-H1组细胞Nm23-H1蛋白表达水平(2.93±0.25),差异有统计学意义(t=5.018,P<0.05)。对照组细胞吸光度(A)值(2.41±0.31)明显高于Nm23-H1组细胞A值(1.89±0.28),差异有统计学意义(t=4.001,P<0.05)。对照组细胞侵袭数量[(102.38±11.37)个]明显高于Nm23-H1组[(78.12±6.90)个],差异有统计学意义(t=3.094,P<0.05)。对照组细胞基质金属蛋白酶(MMP)-3蛋白表达水平(1.29±0.19)明显高于Nm23-H1组(0.69±0.11),差异有统计学意义(t=4.102,P<0.05)。对照组细胞在皮下接种30 d后腋下淋巴结转移数量[(15.22±3.91)个]明显高于Nm23-H1组[(8.73±2.18)个],差异有统计学意义(t=3.094,P<0.05)。对照组细胞N-cadherin和Vimentin蛋白表达水平(1.59±0.22、1.02±0.14)明显高于Nm23-H1组(1.04±0.15、0.61±0.12),差异有统计学意义(t=3.891、3.409,P<0.05)。对照组细胞E-cadherin蛋白表达水平(0.79±0.11)明显高于Nm23-H1组细胞E-cadherin蛋白表达水平(1.51±0.25),差异有统计学意义(t=3.110,P<0.05)。结论:Nm23-H1作为一个转移抑制蛋白,可显著抑制乳腺癌细胞EMT、侵袭和转移。
Objective To investigate the effect of nm23-H1 on epithelial mesenchymal transition(EMT),invasion and metastasis of breast cancer cells.Methods MDA-MB-231 cells were inoculated into 6-well plates,and the control and Nm23-H1 overexpression cell lines were constructed by packaging control and Nm23-H1 lentivirus infection.The proliferation ability of the two groups was analyzed by cell counting kit-8(CCK-8)assay.The protein expression levels of N-cadherin,vimentin and N-cadherin were detected by Western blotting.The invasion ability was analyzed by Transwell and the metastasis ability of cells in vivo was analyzed by the xenograft tumor experiment.SPSS 17.0 statistical software was used to analyze the data and the data were expressed as mean±standard deviation.T test was used to compare the measurement data.Results The expression level of Nm23-H1 in control group(0.87±0.18)was significantly lower than that in Nm23-H1 group(2.93±0.25,t=5.018,P<0.05).The absorbance(A)value of control group(2.41±0.31)was significantly higher than that of Nm23-H1 group(1.89±0.28,t=4.001,P<0.05).The number of invasive cells in the control group(102.38±11.37)was significantly greater than that in the Nm23-H1 group(78.12±6.90,t=3.094,P<0.05).The expression level of matrix metalloproteinase(MMP)-3 protein in control group(1.29±0.19)was significantly higher than that in Nm23-H1 group(0.69±0.11,t=4.102,P<0.05).The number of axillary lymph node metastasis in the control group at 30th day after subcutaneous inoculation(15.22±3.91)was significantly greater than that in the Nm23-H1 group(8.73±2.18,t=3.094,P<0.05).The protein expression levels of N-cadherin and vimentin in control group(1.59±0.22,1.02±0.14)were significantly higher than those in Nm23-H1 group(1.04±0.15,0.61±0.12,t=3.891,3.409,P<0.05).The expression level of E-cadherin in control group(0.79±0.11)was significantly higher than that in Nm23-H1 group(1.51±0.25,t=3.110,P<0.05).Conclusion Nm23-H1as a metastasis suppressor protein,can significantly inhibit EMT,invasion an
作者
赵辉
王雷
冯跃庆
Zhao Hui;Wang Lei;Feng Yueqing(The Second Department of Head,Neck and Breast,Xinxiang Central Hospital,Xinxiang 453000,China;Department of Thyroid and Breast Surgery,the First Affiliated Hospital of Xinxiang Medical College,Xinxiang 453000,China)
出处
《中华实验外科杂志》
CAS
北大核心
2021年第7期1272-1274,共3页
Chinese Journal of Experimental Surgery
关键词
乳腺癌
上皮-间充质转化
侵袭
转移
Breast cancer
Epithelial mesenchymal transition
Invasion
Metastasis
