miR-302b过表达对人舌癌细胞株TSCCa增殖、迁移和侵袭的影响
|
摘要
目的研究miR-302b过表达对人舌癌细胞株TSCCa增殖、迁移和侵袭的影响及其潜在的分子机制。方法用miR-302b-NC和miR-302b mimic转染舌癌细胞TSCCa,命名为miR-302b-NC组和miR-302b mimic组,以未转染的细胞为对照,转染48 h后,采用流式细胞仪检测转染效率,实时荧光定量PCR(qRT-PCR)检测TSCCa细胞中miR-302b的表达情况,分别采用MTT实验、划痕实验和Transwell小室检测TSCCa细胞增殖、迁移和侵袭能力,采用蛋白免疫印迹(Western blot)检测TSCCa细胞中增殖细胞核抗原(PCNA)、基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)表达情况。结果流式细胞仪检测结果显示转染效率达85%以上;转染后,miR-302b mimic组人舌癌细胞TSCCa中miR-302b的表达水平明显高于对照组和miR-302b-NC组(P <0. 05); miR-302b mimic组人舌癌细胞TSCCa增殖率、迁移能力和侵袭能力显著低于对照组和miR-302b-NC组(P <0. 05); miR-302b mimic组人舌癌细胞TSCCa中PCNA、MMP-2、MMP-9蛋白的表达水平明显低于对照组和miR-302b-NC组(P <0. 05)。结论 miR-302b过表达能够抑制人舌癌细胞TSCCa增殖、迁移和侵袭,推测这一作用是通过调控PCNA、MMP-2、MMP-9的表达水平实现的,为舌癌的靶向治疗提供一定的理论依据。
Objective To investigate the effect of miR 302 b overexpression on proliferation,migration and invasion of human tongue cancer cell line TSCCa and its potential molecular mechanism. Methods The tongue cancer cell line TSCCa was transfected with miR 302 b NC and miR 302 b mimic,named as miR 302 b NC group and miR 302 b mimic group,and the non transfected cells were regarded as control group. At 48 hours after transfection,the transfection efficiency was detected by flow cytometry,and the miR 302 b expression in TSCCa cells was detected by quantitative real time fluorescence quantification PCR( qRT PCR). MTT assay,scratch test and transwell chamber test were used to detect the proliferation,migration and invasion ability of TSCCa cells respectively. The expression levels of proliferating cell nuclear antigen( PCNA),matrix metalloproteinase 2( MMP-2) and MMP-9 in TSCCa cells were detected by Western Blot. Results Flow cytometry results showed that transfection efficiency was above 85%. After transfection,the miR 302 b expression level of human tongue cancer cell TSCCa in miR 302 b mimic group was significantly higher than that in the control group and miR 302 b NC group( P <0. 05). The proliferation,migration and invasion abilities of human tongue cancer cell TSCCa in the miR 302 b mimic group were significantly lower than those in control group and miR 302 b NC group( P < 0. 05). The expression levels of PCNA,MMP 2 and MMP 9 of human tongue cancer cell TSCCa in miR 302 b mimic group were significantly lower than those in control group and miR 302 b NC group( P < 0. 05). Conclusion The overexpression of miR 302 b can inhibit the proliferation,migration and invasion of human tongue cancer cell TSCCa,it is speculated that the effects are realized by regulating the expression levels of PCNA,MMP-2 and MMP-9,which provides a theoretical basis for targeted therapy of tongue cancer.
Objective To investigate the effect of miR 302 b overexpression on proliferation,migration and invasion of human tongue cancer cell line TSCCa and its potential molecular mechanism. Methods The tongue cancer cell line TSCCa was transfected with miR 302 b NC and miR 302 b mimic,named as miR 302 b NC group and miR 302 b mimic group,and the non transfected cells were regarded as control group. At 48 hours after transfection,the transfection efficiency was detected by flow cytometry,and the miR 302 b expression in TSCCa cells was detected by quantitative real time fluorescence quantification PCR( qRT PCR). MTT assay,scratch test and transwell chamber test were used to detect the proliferation,migration and invasion ability of TSCCa cells respectively. The expression levels of proliferating cell nuclear antigen( PCNA),matrix metalloproteinase 2( MMP-2) and MMP-9 in TSCCa cells were detected by Western Blot. Results Flow cytometry results showed that transfection efficiency was above 85%. After transfection,the miR 302 b expression level of human tongue cancer cell TSCCa in miR 302 b mimic group was significantly higher than that in the control group and miR 302 b NC group( P <0. 05). The proliferation,migration and invasion abilities of human tongue cancer cell TSCCa in the miR 302 b mimic group were significantly lower than those in control group and miR 302 b NC group( P < 0. 05). The expression levels of PCNA,MMP 2 and MMP 9 of human tongue cancer cell TSCCa in miR 302 b mimic group were significantly lower than those in control group and miR 302 b NC group( P < 0. 05). Conclusion The overexpression of miR 302 b can inhibit the proliferation,migration and invasion of human tongue cancer cell TSCCa,it is speculated that the effects are realized by regulating the expression levels of PCNA,MMP-2 and MMP-9,which provides a theoretical basis for targeted therapy of tongue cancer.
引文
1王芳. 10例舌癌根治术后的皮瓣护理分析.吉林医学,2013,34:1194-1195.
2 刘洋,战德松.舌癌动物模型研究进展.中国实用口腔科杂志,2013,6:311-316.
3 丁慧.阿霉素联合二甲双胍应用对人舌癌细胞SCC-15侵袭迁移能力影响的实验研究.广西医科大学,2017.
4 张翼. miR-302b介导表阿霉素抗骨肉瘤作用及机制研究.武汉大学,2014.
5 崔曼莉,叶文广,王景杰,等. miR-302b在结肠癌组织中的表达及意义.转化医学电子杂志,2017,4:34-36.
6 张鹏江. miR-302b调控食管癌肿瘤相关性炎症信号通路及其机制的初步研究.第四军医大学,2016.
7 陈昌连,梁秀凤.舌癌术后患者与吞咽功能相关的远期生命质量调查分析.中国实用护理杂志,2015,31:2161-2164.
8 徐亚娟,李兆东,高元奇,等. CKS1siRNA联合化疗药物对舌癌Tca8113细胞中CKS1蛋白表达影响的研究.中国实验诊断学,2015,19:1054-1056.
9 Bartel DP. MicroRNAs:genomics,biogenesis,mechanism,and function.Cell,2004,116:281-297.
10 田晓琳,杨臻,王建英,等.微小RNA与肿瘤的关系.癌症进展,2016,14:22-25.
11 李芳,李玉凤,张景华.微小RNA通过调控肿瘤血管形成影响肿瘤转移.基因组学与应用生物学,2015,34:2332-2337.
12 Rottiers V,Nr AM. MicroRNAs in Metabolism and Metabolic Disorders.Nature Reviews Molecular Cell Biology,2012,13:239-250.
13 Krol J,Loedige I,Filipowicz W. The widespread regulation of microRNA biogenesis,function and decay. Nature Reviews Genetics,2010,11:597-610.
14 万勇,吴心愿. PCNA和EMMPRIN在食管鳞癌中的表达及其临床意义.肿瘤,2016,31:141-144.
15 郑华峰,陶晶,张斌,等. MiR-495通过下调PCNA抑制人血管内皮细胞增殖的作用研究.中国心血管病研究,2016,14:374-379.
16 高红强,刘静,李志强,等. MiR-199a-3p对大鼠肝细胞增殖的抑制作用.昆明医科大学学报,2017,38:10-16.
17 黎进,马炬明.基质金属蛋白酶-2、9在胃癌中的研究进展.中国肿瘤,2015,24:403-407.
18 Mou L,Kang Y,Zhou Y,et al. Neurokinin-1 receptor directly mediates glioma cell migration by up-regulation of matrix metalloproteinase-2(MMP-2)and membrane type 1-matrix metalloproteinase(MT1-MMP). Journal of Biological Chemistry,2013,288:306-318.
19 任蕾,喻学洲,周永庆. MMP-9和CD44V6在舌癌中的表达及与淋巴结转移的关系.贵阳医学院学报,2015,40:1356-1358.
2 刘洋,战德松.舌癌动物模型研究进展.中国实用口腔科杂志,2013,6:311-316.
3 丁慧.阿霉素联合二甲双胍应用对人舌癌细胞SCC-15侵袭迁移能力影响的实验研究.广西医科大学,2017.
4 张翼. miR-302b介导表阿霉素抗骨肉瘤作用及机制研究.武汉大学,2014.
5 崔曼莉,叶文广,王景杰,等. miR-302b在结肠癌组织中的表达及意义.转化医学电子杂志,2017,4:34-36.
6 张鹏江. miR-302b调控食管癌肿瘤相关性炎症信号通路及其机制的初步研究.第四军医大学,2016.
7 陈昌连,梁秀凤.舌癌术后患者与吞咽功能相关的远期生命质量调查分析.中国实用护理杂志,2015,31:2161-2164.
8 徐亚娟,李兆东,高元奇,等. CKS1siRNA联合化疗药物对舌癌Tca8113细胞中CKS1蛋白表达影响的研究.中国实验诊断学,2015,19:1054-1056.
9 Bartel DP. MicroRNAs:genomics,biogenesis,mechanism,and function.Cell,2004,116:281-297.
10 田晓琳,杨臻,王建英,等.微小RNA与肿瘤的关系.癌症进展,2016,14:22-25.
11 李芳,李玉凤,张景华.微小RNA通过调控肿瘤血管形成影响肿瘤转移.基因组学与应用生物学,2015,34:2332-2337.
12 Rottiers V,Nr AM. MicroRNAs in Metabolism and Metabolic Disorders.Nature Reviews Molecular Cell Biology,2012,13:239-250.
13 Krol J,Loedige I,Filipowicz W. The widespread regulation of microRNA biogenesis,function and decay. Nature Reviews Genetics,2010,11:597-610.
14 万勇,吴心愿. PCNA和EMMPRIN在食管鳞癌中的表达及其临床意义.肿瘤,2016,31:141-144.
15 郑华峰,陶晶,张斌,等. MiR-495通过下调PCNA抑制人血管内皮细胞增殖的作用研究.中国心血管病研究,2016,14:374-379.
16 高红强,刘静,李志强,等. MiR-199a-3p对大鼠肝细胞增殖的抑制作用.昆明医科大学学报,2017,38:10-16.
17 黎进,马炬明.基质金属蛋白酶-2、9在胃癌中的研究进展.中国肿瘤,2015,24:403-407.
18 Mou L,Kang Y,Zhou Y,et al. Neurokinin-1 receptor directly mediates glioma cell migration by up-regulation of matrix metalloproteinase-2(MMP-2)and membrane type 1-matrix metalloproteinase(MT1-MMP). Journal of Biological Chemistry,2013,288:306-318.
19 任蕾,喻学洲,周永庆. MMP-9和CD44V6在舌癌中的表达及与淋巴结转移的关系.贵阳医学院学报,2015,40:1356-1358.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |