miRNA-186调控CDK6和IL-10表达抑制乳腺癌MCF-7细胞生长与转移机制的研究
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摘要
目的探讨miR-186介导的CDK6和IL-10表达对乳腺癌MCF-7细胞增殖、凋亡及侵袭的影响。方法用已构建的miR-186 mimic质粒转染乳腺癌MCF-7细胞,荧光显微镜下观察转染效率,Western blot技术检测重组质粒对CDK6和IL-10表达的影响,光学显微镜下观察细胞生长群落,分别用MTT法、流式细胞术和Transwell小室法测定重组质粒转染对细胞增殖、凋亡周期及侵袭能力的影响。结果 miR-186转染效率良好,与non-targeting组比较,在转染miR-186后,可明显抑制乳腺癌MCF-7细胞的增殖(P<0.01),细胞生长群落分析结果显示,miR-186组细胞群落数目明显低于non-targeting组细胞(P<0.01),细胞周期结果显示,miR-186组的G0/G1和S期百分比明显的低于non-targeting组(P<0.05),G2/M期百分比明显的高于non-targeting组(P<0.05),Transwell检测结果显示,miR-186组乳腺癌MCF-7细胞侵袭细胞数量(81.00±2.00)个明显少于non-targeting组(125.00±3.00)个,miR-186转染后,CDK6和IL-10在miR-186组乳腺癌MCF-7细胞中的表达率明显低于non-targeting组。结论 miR-186可下调乳腺癌MCF-7细胞中CDK6和IL-10的表达,可有效抑制乳腺癌MCF-7细胞的增殖、侵袭,miR-186可能成为一个潜在的乳腺癌基因治疗靶标。
This study was designed to investigate the effects of miR-186-mediated CDK6 and Il-10 expression on the proliferation, apoptosis and invasion of breast cancer MCF-7 cells. miR-186 mimic plasmid was adopted to transfect MCF-7 cells, and the transfection efficiency was observed with fluorescence microscope.Western blot was used to detect recombinant plasmid expression of CDK6 and IL-10, while optical microscope was used to observe th cell growth. The effects of recombinant plasmid transfection on the proliferation, apoptosis, and invasive ability of MCF-7 cells were evaluated by MTT method, flow cytometry and Transwell Chambers method.The data showed that miR-186 transfection could significantly inhibit the proliferation of breast cancer MCF-7 cells(P<0.01). The results of cell growth community analysis showed that the number of cell communities in the miR-186 group was significantly lower than that in the non-targeting group(P<0.01). The percentage of G2/M stage cells was significantly higher than that of the non-targeting group(P<0.05). Transwell test results showed that the number of breast cancer MCF-7 cells in the miR-186 group(81.00±2.00) was significantly lower than that in the non-targeting group(125.00±3.00). After transfection with miR-186,the expression rate of CDK6 and IL-10 in breast cancer MCF-7 cells of the miR-186 group was significantly lower than that the non-targeting group. In conclusion, miR-186 can down-regulate the expression of CDK6 and IL-10 in breast cancer MCF-7 cells and effectively inhibit the proliferation and invasion of breast cancer MCF-7 cells, thus miR-186 may become a potential target for breast cancer gene therapy.
This study was designed to investigate the effects of miR-186-mediated CDK6 and Il-10 expression on the proliferation, apoptosis and invasion of breast cancer MCF-7 cells. miR-186 mimic plasmid was adopted to transfect MCF-7 cells, and the transfection efficiency was observed with fluorescence microscope.Western blot was used to detect recombinant plasmid expression of CDK6 and IL-10, while optical microscope was used to observe th cell growth. The effects of recombinant plasmid transfection on the proliferation, apoptosis, and invasive ability of MCF-7 cells were evaluated by MTT method, flow cytometry and Transwell Chambers method.The data showed that miR-186 transfection could significantly inhibit the proliferation of breast cancer MCF-7 cells(P<0.01). The results of cell growth community analysis showed that the number of cell communities in the miR-186 group was significantly lower than that in the non-targeting group(P<0.01). The percentage of G2/M stage cells was significantly higher than that of the non-targeting group(P<0.05). Transwell test results showed that the number of breast cancer MCF-7 cells in the miR-186 group(81.00±2.00) was significantly lower than that in the non-targeting group(125.00±3.00). After transfection with miR-186,the expression rate of CDK6 and IL-10 in breast cancer MCF-7 cells of the miR-186 group was significantly lower than that the non-targeting group. In conclusion, miR-186 can down-regulate the expression of CDK6 and IL-10 in breast cancer MCF-7 cells and effectively inhibit the proliferation and invasion of breast cancer MCF-7 cells, thus miR-186 may become a potential target for breast cancer gene therapy.
引文
[1]张建兴.多种影像学方法在中国乳腺癌筛查中的应用[J].实用医学杂志,2017,33(9):1365-1368.
[2]郑莹.中国乳腺癌患者生活方式指南[J].中华外科杂志,2017,39(2):124-128.
[3]宁莉,魏殿军.急性肾损伤相关miRNA的研究进展[J].山东医药,2017,57(14):110-112.
[4]陈梦圆,王艳林,黄利鸣.miRNA与宫颈癌相关性的研究进展[J].现代妇产科进展,2017,26(1):73-75.
[5]Huang Y,Shen X J,Zou Q,et al.Biological functions of MicroRNAs[J].J Physiol Biochem,2010,36(6):684-689.
[6]陈灿斌,刘小敏,梁晓秋.miR-186靶向PIG11基因抑制胃癌MKN-28细胞侵袭的作用[J].中国现代医药杂志,2015,17(1):1-4.
[7]孙艳,齐政,王德华,等.miR-186通过靶定CEA抑制宫颈癌细胞转移[J].天津医科大学学报,2016,22(4):288-290.
[8]张国新,徐新伟,孟斌,等.miR-186-5p通过靶向调控PTTG1抑制肺腺癌细胞的上皮-间质转化[J].中国生物化学与分子生物学报,2017,33(4):380-385.
[9]马俊骥,郭平,郭昱,等.血浆miR-92a-3p和miR-204-5p及miR-186-5p诊断胆管癌的价值研究[J].中国全科医学,2017,20(30):3736-3739.
[10]宋丹,王孜尧,黄平.微RNA-186在人肝细胞肝癌中的表达及作用[J].肿瘤,2017,37(7):750-761.
[11]Lu S,Wang MS,Chen PJ,et al.miRNA-186 inhibits prostate cancer cell proliferation and tumor growth by targeting YY1 and CDK6[J].Exp Therap Med,2017,13(6):3309-3314.
[12]李卿,王红梅,刘燕玲,等.IL-10(-1082A/G)基因多态性和乳腺癌的相关性:Meta分析[J].现代生物医学进展,2017,17(5):820-825.
[13]郑丽,赵敏,昂琳,等.乳腺癌中BCL-6和ZEB2的表达及其意义[J].临床与实验病理学杂志,2017,33(1):50-54.
[14]杨浚沨,王龙强,李海,等.乳腺癌循环miRNA生物标志物的筛选及验证[J].中国普通外科杂志,2015,24(5):696-700.
[15]杨丽,韩娜,石磊,等.长链非编码RNA在胶质母细胞瘤中异常表达的研究进展[J].实用医学杂志,2017,33(21):3670-3673.
[16]常永梅,王明智,江小运,等.miR-145高甲基化在非小细胞肺癌组织中检测的意义[J].实用医学杂志,2016,32(16):2671-2673.
[17]李永文,张洪兵,李颖,等.免疫抑制剂FK506对肺癌细胞增殖和迁移的作用研究[J].中国肺癌杂志,2017,20(7):446-451.
[18]刘静,江叶,闵敏,等.P21及CDK6与宫颈鳞状细胞癌的关系[J].重庆医学,2017,46(23):3231-3233.
[19]董一楠,张新伟,魏枫.CDK6导致肿瘤的机制研究进展[J].中国肿瘤临床,2015,42(19):973-977.
[20]Yuan S,Qu Y,Dang S,et al.MiR-145 inhibits oral squamous cell carcinoma(OSCC)cell growth by targeting c-Myc and Cdk6[J].Cancer Cell International,2013,13(1):51-60.
[21]李婷婷,王福玲,张桂萍,等.miR-145和IL-10在子宫内膜样腺癌的表达及其相关性[J].免疫学杂志,2017,33(1):89-92.
[2]郑莹.中国乳腺癌患者生活方式指南[J].中华外科杂志,2017,39(2):124-128.
[3]宁莉,魏殿军.急性肾损伤相关miRNA的研究进展[J].山东医药,2017,57(14):110-112.
[4]陈梦圆,王艳林,黄利鸣.miRNA与宫颈癌相关性的研究进展[J].现代妇产科进展,2017,26(1):73-75.
[5]Huang Y,Shen X J,Zou Q,et al.Biological functions of MicroRNAs[J].J Physiol Biochem,2010,36(6):684-689.
[6]陈灿斌,刘小敏,梁晓秋.miR-186靶向PIG11基因抑制胃癌MKN-28细胞侵袭的作用[J].中国现代医药杂志,2015,17(1):1-4.
[7]孙艳,齐政,王德华,等.miR-186通过靶定CEA抑制宫颈癌细胞转移[J].天津医科大学学报,2016,22(4):288-290.
[8]张国新,徐新伟,孟斌,等.miR-186-5p通过靶向调控PTTG1抑制肺腺癌细胞的上皮-间质转化[J].中国生物化学与分子生物学报,2017,33(4):380-385.
[9]马俊骥,郭平,郭昱,等.血浆miR-92a-3p和miR-204-5p及miR-186-5p诊断胆管癌的价值研究[J].中国全科医学,2017,20(30):3736-3739.
[10]宋丹,王孜尧,黄平.微RNA-186在人肝细胞肝癌中的表达及作用[J].肿瘤,2017,37(7):750-761.
[11]Lu S,Wang MS,Chen PJ,et al.miRNA-186 inhibits prostate cancer cell proliferation and tumor growth by targeting YY1 and CDK6[J].Exp Therap Med,2017,13(6):3309-3314.
[12]李卿,王红梅,刘燕玲,等.IL-10(-1082A/G)基因多态性和乳腺癌的相关性:Meta分析[J].现代生物医学进展,2017,17(5):820-825.
[13]郑丽,赵敏,昂琳,等.乳腺癌中BCL-6和ZEB2的表达及其意义[J].临床与实验病理学杂志,2017,33(1):50-54.
[14]杨浚沨,王龙强,李海,等.乳腺癌循环miRNA生物标志物的筛选及验证[J].中国普通外科杂志,2015,24(5):696-700.
[15]杨丽,韩娜,石磊,等.长链非编码RNA在胶质母细胞瘤中异常表达的研究进展[J].实用医学杂志,2017,33(21):3670-3673.
[16]常永梅,王明智,江小运,等.miR-145高甲基化在非小细胞肺癌组织中检测的意义[J].实用医学杂志,2016,32(16):2671-2673.
[17]李永文,张洪兵,李颖,等.免疫抑制剂FK506对肺癌细胞增殖和迁移的作用研究[J].中国肺癌杂志,2017,20(7):446-451.
[18]刘静,江叶,闵敏,等.P21及CDK6与宫颈鳞状细胞癌的关系[J].重庆医学,2017,46(23):3231-3233.
[19]董一楠,张新伟,魏枫.CDK6导致肿瘤的机制研究进展[J].中国肿瘤临床,2015,42(19):973-977.
[20]Yuan S,Qu Y,Dang S,et al.MiR-145 inhibits oral squamous cell carcinoma(OSCC)cell growth by targeting c-Myc and Cdk6[J].Cancer Cell International,2013,13(1):51-60.
[21]李婷婷,王福玲,张桂萍,等.miR-145和IL-10在子宫内膜样腺癌的表达及其相关性[J].免疫学杂志,2017,33(1):89-92.