摘要
目的 观察过表达叉头框转录因子C1(FOXC1)对人非小细胞肺癌H1299细胞生物学行为及基质金属蛋白酶2(MMP-2)的影响。方法 将H1299细胞分为观察组与对照组,分别转染FOXC1目的基因质粒和空载体质粒。转染48 h后收集细胞,采用实时荧光定量PCR法和Western blotting法检测细胞FOXC1 mRNA和蛋白表达,MTT法检测培养1~8 d的细胞增殖能力,平板克隆实验检测细胞克隆形成能力,Transwell细胞侵袭实验检测穿过基底膜的细胞数,Western blotting法检测MMP-2蛋白相对表达量。结果 观察组细胞FOXC1 mRNA及蛋白相对表达量均高于对照组(P均<0.05)。两组培养1~4 d细胞增殖能力比较差异均无统计学意义(P均>0.05),观察组培养5~8 d细胞增殖能力均高于对照组(P均<0.05)。观察组细胞克隆形成率、穿过基底膜的细胞数、MMP-2蛋白相对表达量均高于对照组(P均<0.05)。结论 过表达的FOXC1可能通过上调MMP-2表达而促进非小细胞肺癌H1299细胞增殖及侵袭。
Objective To observe the effect of FOXC1 over-expression on the biological behavior and matrix metalloproteinase 2(MMP-2)of human non-small-cell lung cancer(NSCLC)cell line H1299.Methods H1299 cells in the logarithmic growth phase were divided into the observation group and control group,which were transfected with FOXC1 target gene plasmid and empty vector plasmid,respectively.After 48 hours of transfection,FOXC1 mRNA and protein expression levels were detected by real-time fluorescent quantitative PCR and Western blotting,cell proliferation was detected by MTT,cell clone formation was detected by plate blotting,the number of cells passing through basement membrane was detected by Transwell cell invasion test,and MMP-2 protein expression was detected by Western blotting.Results The relative expression levels of FOXC1 mRNA and protein in the observation group were higher than those in the control group(both P<0.05).There was no significant difference in the cell proliferation between the two groups at 1-4 days after culture(P>0.05).The cell proliferation of the observation group was higher than that of the control group at 5-8 days after culture(P<0.05).The rate of cell clone formation,the number of cells passing through the basement membrane,and the relative expression of MMP-2 protein in the observation group were all higher than those in the control group(all P<0.05).Conclusion The over-expression of FOXC1 can enhance the proliferation and invasion abilities of H1299 cells through up-regulating the expression of MMP-2.
作者
韩乐
赵征
宋养荣
雷宝霞
陈文娟
HAN Le;ZHAO Zheng;SONG Yangrong;LEI Baoxia;CHEN Wenjuan(Shaanxi Provincial Cancer Hospital,Xi'an 710061,China)
出处
《山东医药》
CAS
2019年第35期12-15,共4页
Shandong Medical Journal
基金
陕西省自然科学基础研究计划(2019JM538)
陕西省卫健委卫生健康科研基金资助项目(2018D032)
