长链非编码RNA MIR200CHG结合尿苷二磷酸葡萄糖脱氢酶促进乳腺癌细胞的侵袭  

Long noncoding RNA MIR200CHG binds uridine diphosphate glucose dehydrogenase to promote breast cancer cell invasion

作　　者：成道福[1] 韦达[2] 唐莉[1] Cheng Daofu;Wei Da;Tang Li(Department of Clinical Laboratory,Jiangsu Cancer Hospital,Jiangsu Institute of Cancer Research,the Affiliated Cancer Hospital of Nanjing Medical University,Nanjing 210009,China;Department of General Surgery,Jiangsu Cancer Hospital,Jiangsu Institute of Cancer Research,the Affiliated Cancer Hospital of Nanjing Medical University,Nanjing 210009,China)

机构地区：[1]江苏省肿瘤医院江苏省肿瘤防治研究所南京医科大学附属肿瘤医院检验科,210009 [2]江苏省肿瘤医院江苏省肿瘤防治研究所南京医科大学附属肿瘤医院普外科,210009

出　　处：《中华实验外科杂志》2022年第9期1677-1679,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(81802898);江苏省自然科学基金(BK20181090)。

摘　　要：目的观察长链非编码RNA微小RNA-200c和微小RNA-141宿主基因(MIR200CHG)结合尿苷二磷酸葡萄糖脱氢酶(UGDH)对乳腺癌细胞侵袭的影响。方法采用RNA pull-down实验结合质谱分析MIR200CHG与UGDH之间的相互作用;用实时定量聚合酶链反应和蛋白质印迹分析MIR200CHG表达下调对UGDH mRNA和蛋白的影响;用Transwell实验分析MIR200CHG结合UGDH对乳腺癌细胞侵袭能力的影响。组间比较采用配对t检验。结果UGDH蛋白通过肽段VLIGGDETPEGQR和LAANAFLAQR与MIR200CHG结合,且仅在MIR200CHG正义链结合的蛋白液中检测到UGDH蛋白。下调MCF7细胞中的MIR200CHG使实验组中UGDH蛋白表达低于阴性对照组(0.38±0.09比1.00±0.03,t=11.320,P<0.01);挽救MCF7-sh细胞中的UGDH表达使实验组中发生侵袭的细胞比例高于阴性对照组(5.34±1.06比1.00±0.20,t=6.967,P<0.01),同时实验组中基质金属蛋白酶(MMP)-2、MMP-9和波形蛋白表达高于阴性对照组,分别为(1.96±0.23比1.01±0.04,t=7.064,P<0.01)、(1.75±0.17比1.00±0.05,t=7.330,P<0.01)和(1.63±0.17比1.02±0.02,t=6.158,P<0.01)。结论MIR200CHG可能通过结合UGDH蛋白促进乳腺癌细胞的侵袭。Objective To explore the effects of long noncoding RNA microRNA-200c and microRNA-141 host gene(MIR200CHG)combined with uridine diphospho-glucose dehydrogenase(UGDH)on breast cancer cell invasion.Methods The interactions between MIR200CHG and UGDH were analyzed by RNA pull-down experiment and mass spectrometry.The effects of down-regulation of MIR200CHG on UGDH mRNA and protein were analyzed by real-time quantitative polymerase chain reaction and Western blotting.The effect of MIR200CHG combined with UGDH on the invasion ability of breast cancer cells was analyzed by Transwell experiment.The data were processed with GraphPad Prism 8 software,and the paired t-test was used for comparison between groups.Results UGDH protein bound to MIR200CHG via peptides VLIGGDETPEGQR and LAANAFLAQR.UGDH protein was only detected in the protein solution bound by the sense strand of MIR200CHG.After down-regulation of MIR200CHG in MCF7 cells,the expression of UGDH protein in the experimental groups was lower than that in the negative control group(0.38±0.09 vs.1.00±0.03,t=11.320,P<0.01).Rescue of UGDH expression in MCF7-sh cells increased the proportion of invasive cells in the experimental groups as compared with the negative control group(5.34±1.06 vs.1.00±0.20,t=6.967,P<0.01).The expression of matrix metalloproteinase(MMP)-2,MMP-9 and vimentin in the experimental groups was higher than that in the negative control groups(1.96±0.23 vs.1.01±0.04,t=7.064,P<0.01;1.75±0.17 vs.1.00±0.05,t=7.330,P<0.01;1.63±0.17 vs.1.02±0.02,t=6.158,P<0.01).Conclusion MIR200CHG may promote the invasion of breast cancer cells by binding to UGDH.

关 键 词：乳腺癌 长链非编码RNA 尿苷二磷酸葡萄糖脱氢酶 侵袭 

分 类 号：R737.9[医药卫生—肿瘤]