槐耳清膏对乳腺癌细胞microRNA表达谱的影响
|
摘要
目的探讨槐耳清膏对乳腺癌MCF-7和MBA-MD-231细胞系微小RNA(microRNA)表达谱的影响,为槐耳清膏治疗乳腺癌的分子机制提供依据。方法选择乳腺癌细胞MCF-7及MDA-MB-231,分为槐耳清膏处理组和对照组。以两组MCF-7及MBA-MD-231细胞系总RNA为模板,应用半定量PCR方法(qRT-PCR)反转录后得到c DNA模板,利用特异性定量PCR引物,凝胶电泳,对microRNA表达量进行分析,以荧光信号强度的比值log2(ratio)得到差异性表达的microRNA。结果槐耳清膏使乳腺癌MCF-7细胞中表达上调的microRNA有6个,表达下调的有21个;使MDA-MB-231细胞中表达上调的microRNA有3个,表达下调的有24个。其中表达上调显著的microRNA有MCF-7细胞系中的miR-7-5p、miR-21-5p、miR-126-3p和MBA-MD-231细胞系中的miR-10b-5p、miR-21-5p。结论槐耳清膏使乳腺癌MCF-7和MDA-MB-231细胞中miR-7-5p、miR-21-5p、miR-126-3p与miR-10b-5p、miR-21-5p表达显著上调,其异常表达的microRNA有可能作为乳腺癌药物治疗潜在的新的靶点。
Objective To explore the effect of Huaier on the expression profile of microRNAs in the breast cancer MCF-7 and MBA-MD-231 cell lines,and to provide a molecular mechanism for the treatment of breast cancer with Huaier.Methods MCF-7 and MDA-MB-231 breast cancer cells were selected and were divided into two groups: the treatment group and the control group. By using the total RNA of two sets of MCF-7 and MBA-MD-231 cell lines as templates,the c DNA template was obtained by qRT-PCR after reverse transcription. The microRNA expression was analyzed by specific PCR primers and gel electrophoresis,and we obtained the differentially expressed microRNAs by the ratio of fluorescence intensity to log2( ratio). Results Six microRNAs in the breast cancer MCF-7 cells were up-regulated and 21 microRNAs were down-regulated by Huaier; three microRNAs in breast cancer MDA-MB-231 cells were up-regulated and 24 microRNAs were down-regulated by Huaier. The significantly up-regulated microRNAs are miR-7-5 p,miR-21-5 p,and miR-126-3 p in MCF-7 cells,and miR-10 b-5 p and miR-21-5 p in the MBA-MD-231 cells. Conclusion Huaier significantly up-regulates the expression of microRNAs in breast cancer MCF-7 and MDA-MB-231 cells,including miR-7-5 p,miR-21-5 p,miR-126-3 p,miR-10 b-5 p and miR-21-5 p,and the abnormal expression of microRNAs may be a potential new target for the treatment of breast cancer.
Objective To explore the effect of Huaier on the expression profile of microRNAs in the breast cancer MCF-7 and MBA-MD-231 cell lines,and to provide a molecular mechanism for the treatment of breast cancer with Huaier.Methods MCF-7 and MDA-MB-231 breast cancer cells were selected and were divided into two groups: the treatment group and the control group. By using the total RNA of two sets of MCF-7 and MBA-MD-231 cell lines as templates,the c DNA template was obtained by qRT-PCR after reverse transcription. The microRNA expression was analyzed by specific PCR primers and gel electrophoresis,and we obtained the differentially expressed microRNAs by the ratio of fluorescence intensity to log2( ratio). Results Six microRNAs in the breast cancer MCF-7 cells were up-regulated and 21 microRNAs were down-regulated by Huaier; three microRNAs in breast cancer MDA-MB-231 cells were up-regulated and 24 microRNAs were down-regulated by Huaier. The significantly up-regulated microRNAs are miR-7-5 p,miR-21-5 p,and miR-126-3 p in MCF-7 cells,and miR-10 b-5 p and miR-21-5 p in the MBA-MD-231 cells. Conclusion Huaier significantly up-regulates the expression of microRNAs in breast cancer MCF-7 and MDA-MB-231 cells,including miR-7-5 p,miR-21-5 p,miR-126-3 p,miR-10 b-5 p and miR-21-5 p,and the abnormal expression of microRNAs may be a potential new target for the treatment of breast cancer.
引文
[1]Siegel RL,Miller KD,Jemal A.Cancer statistics,2015[J].CACancer J Clin,2015,65(1):5-29.
[2]Lux MP,Janni W,Hartkopf AD,et al.Update breast cancer 2017-implementation of novel therapies[J].Geburtshilfe Frauenheilkd,2017,77(12):1281-1290.
[3]冯雪松,鲁明骞.miRNA与乳腺癌治疗抵抗的关系及机制研究进展[J].实用医学杂志,2017,33(14):2412-2414.
[4]Ling H,Krassnig L,Bullock MD,et al.MicroRNAs in testicular cancer diagnosis and prognosis[J].Urol Clin North Am,2016,43(1):127-134.
[5]Zhang J,Xiao X,Liu J.The role of circulating miRNAs in multiple myeloma[J].Sci China Life Sci,2015,58(12):1262-1269.
[6]Boeri M,Verri C,Conte D,et al.MicroRNA signatures in tissues and plasma predict development and prognosis of computed tomography detected lung cancer[J].Proc Natl Acad Sci USA,2011,108(9):3713-3718.
[7]Brase JC,Wuttig D,Kuner R,et al.Serum microRNAs as non-invasive biomarkers for cancer[J].Mol Cancer,2010,9:306.
[8]鲁明骞,冯雪松,孔庆志,等.槐耳清膏抑制MCF-7细胞裸鼠成瘤性实验研究[J].东南大学学报(医学版),2018,37(1):18-21.
[9]Luo Z,Hu X,Xiong H,et al.A polysaccharide from Huaier induced apoptosis in MCF-7 breast cancer cells via down-regulation of MTDH protein[J].Carbohydr Polym,2016,151:1027-1033.
[10]Li Y,Qi W,Song X,et al.Huaier extract suppresses breast cancer via regulating tumor-associated macrophages[J].Sci Rep,2016,6:20049.
[11]Wang X,Qi W,Li Y,et al.Huaier extract induces autophagic cell death by inhibiting the mtor/s6k pathway in breast cancer cells[J].PLo S One,2015,10(7):e131771.
[12]Jiang C,Chen X,Alattar M,et al.MicroRNAs in tumorigenesis,metastasis,diagnosis and prognosis of gastric cancer[J].Cancer Gene Ther,2015,22(6):291-301.
[13]Li C,Zhang K,Chen J,et al.MicroRNAs as regulators and mediators of forkhead box transcription factors function in human cancers[J].Oncotarget,2017,8(7):12433-12450.
[14]黄幼生,解娜,张艺馨,等.胃癌组织microRNA表达谱检测及生物信息学分析[J].临床与实验病理学杂志,2017,33(6):591-595.
[15]He M,Zhou W,Li C,et al.MicroRNAs,DNA damage response,and cancer treatment[J].Int J Mol Sci,2016,17(12).
[16]周学,杜宜兰,金萍,等.癌症相关microRNA与靶基因的生物信息学分析[J].遗传,2015(09):855-864.
[17]刘冬冬.乳腺癌血清microRNA表达谱差异分析及其功能验证[D].广州:广州中医药大学,2016.
[18]Sochor M,Basova P,Pesta M,et al.Oncogenic microRNAs:miR-155,miR-19a,miR-181b,and miR-24 enable monitoring of early breast cancer in serum[J].BMC Cancer,2014,14:448.
[19]Paszek S,Gablo N,Barnas E,et al.Dysregulation of microRNAs in triple-negative breast cancer[J].Ginekol Pol,2017,88(10):530-536.
[20]Jin FP,Zhang M.Progress of experimental researches on Chinese herbal compounds for inducing tumor cell apoptosis[J].Chin J Integr Med,2010,16(6):565-571.
[21]黄涛,孔庆志,卢宏达,等.槐耳清膏对耐顺铂人肺腺癌细胞系A549DDP逆转的实验研究[J].中国药师,2002,5(9):517-521.
[22]鲁明骞,冯雪松,孔庆志,等.槐耳清膏抑制MCF-细胞裸鼠成瘤性实验研究[J].东南大学学报:医学版,2018,37(1):18-21.
[23]林维佳,郑琳,代建波,等.靶向EGFR的miR-7-5p对人乳腺癌细胞增殖和侵袭的影响[J].中华肿瘤防治杂志,2015,22(15):1176-1176.
[24]蔡洲,黄致远,张建新.miR-126-3p在甲状腺癌中的表达及其功能研究[J].中国免疫学杂志,2017,33(4):584-592.
[25]蔡如玉,张金玲,唐丹燕,等.血清miR-21-5p在子宫内膜样腺癌中的表达及意义[J].山西医科大学学报,2018,49(6):618-621.
[26]Bertoli G,Cava C,Castiglioni I.MicroRNAs:new biomarkers for diagnosis,prognosis,therapy prediction and therapeutic tools for breast cancer[J].Theranostics,2015,5(10):1122-1143.
[27]Chen Q,Hu H,Jiao D,et al.miR-126-3p and miR-451a correlate with clinicopathological features of lung adenocarcinoma:The underlying molecular mechanisms[J].Oncol Rep,2016,36(2):909-917.
[28]Park HK,Jo W,Choi HJ,et al.Time-course changes in the ex pression levels of miR-122,-155,and-21 as markers of liver cell damage,inflamation,and regeneration in acetaminophen-induced liver injury in rats[J].Vet Sci,2016,17(1):45-51.
[2]Lux MP,Janni W,Hartkopf AD,et al.Update breast cancer 2017-implementation of novel therapies[J].Geburtshilfe Frauenheilkd,2017,77(12):1281-1290.
[3]冯雪松,鲁明骞.miRNA与乳腺癌治疗抵抗的关系及机制研究进展[J].实用医学杂志,2017,33(14):2412-2414.
[4]Ling H,Krassnig L,Bullock MD,et al.MicroRNAs in testicular cancer diagnosis and prognosis[J].Urol Clin North Am,2016,43(1):127-134.
[5]Zhang J,Xiao X,Liu J.The role of circulating miRNAs in multiple myeloma[J].Sci China Life Sci,2015,58(12):1262-1269.
[6]Boeri M,Verri C,Conte D,et al.MicroRNA signatures in tissues and plasma predict development and prognosis of computed tomography detected lung cancer[J].Proc Natl Acad Sci USA,2011,108(9):3713-3718.
[7]Brase JC,Wuttig D,Kuner R,et al.Serum microRNAs as non-invasive biomarkers for cancer[J].Mol Cancer,2010,9:306.
[8]鲁明骞,冯雪松,孔庆志,等.槐耳清膏抑制MCF-7细胞裸鼠成瘤性实验研究[J].东南大学学报(医学版),2018,37(1):18-21.
[9]Luo Z,Hu X,Xiong H,et al.A polysaccharide from Huaier induced apoptosis in MCF-7 breast cancer cells via down-regulation of MTDH protein[J].Carbohydr Polym,2016,151:1027-1033.
[10]Li Y,Qi W,Song X,et al.Huaier extract suppresses breast cancer via regulating tumor-associated macrophages[J].Sci Rep,2016,6:20049.
[11]Wang X,Qi W,Li Y,et al.Huaier extract induces autophagic cell death by inhibiting the mtor/s6k pathway in breast cancer cells[J].PLo S One,2015,10(7):e131771.
[12]Jiang C,Chen X,Alattar M,et al.MicroRNAs in tumorigenesis,metastasis,diagnosis and prognosis of gastric cancer[J].Cancer Gene Ther,2015,22(6):291-301.
[13]Li C,Zhang K,Chen J,et al.MicroRNAs as regulators and mediators of forkhead box transcription factors function in human cancers[J].Oncotarget,2017,8(7):12433-12450.
[14]黄幼生,解娜,张艺馨,等.胃癌组织microRNA表达谱检测及生物信息学分析[J].临床与实验病理学杂志,2017,33(6):591-595.
[15]He M,Zhou W,Li C,et al.MicroRNAs,DNA damage response,and cancer treatment[J].Int J Mol Sci,2016,17(12).
[16]周学,杜宜兰,金萍,等.癌症相关microRNA与靶基因的生物信息学分析[J].遗传,2015(09):855-864.
[17]刘冬冬.乳腺癌血清microRNA表达谱差异分析及其功能验证[D].广州:广州中医药大学,2016.
[18]Sochor M,Basova P,Pesta M,et al.Oncogenic microRNAs:miR-155,miR-19a,miR-181b,and miR-24 enable monitoring of early breast cancer in serum[J].BMC Cancer,2014,14:448.
[19]Paszek S,Gablo N,Barnas E,et al.Dysregulation of microRNAs in triple-negative breast cancer[J].Ginekol Pol,2017,88(10):530-536.
[20]Jin FP,Zhang M.Progress of experimental researches on Chinese herbal compounds for inducing tumor cell apoptosis[J].Chin J Integr Med,2010,16(6):565-571.
[21]黄涛,孔庆志,卢宏达,等.槐耳清膏对耐顺铂人肺腺癌细胞系A549DDP逆转的实验研究[J].中国药师,2002,5(9):517-521.
[22]鲁明骞,冯雪松,孔庆志,等.槐耳清膏抑制MCF-细胞裸鼠成瘤性实验研究[J].东南大学学报:医学版,2018,37(1):18-21.
[23]林维佳,郑琳,代建波,等.靶向EGFR的miR-7-5p对人乳腺癌细胞增殖和侵袭的影响[J].中华肿瘤防治杂志,2015,22(15):1176-1176.
[24]蔡洲,黄致远,张建新.miR-126-3p在甲状腺癌中的表达及其功能研究[J].中国免疫学杂志,2017,33(4):584-592.
[25]蔡如玉,张金玲,唐丹燕,等.血清miR-21-5p在子宫内膜样腺癌中的表达及意义[J].山西医科大学学报,2018,49(6):618-621.
[26]Bertoli G,Cava C,Castiglioni I.MicroRNAs:new biomarkers for diagnosis,prognosis,therapy prediction and therapeutic tools for breast cancer[J].Theranostics,2015,5(10):1122-1143.
[27]Chen Q,Hu H,Jiao D,et al.miR-126-3p and miR-451a correlate with clinicopathological features of lung adenocarcinoma:The underlying molecular mechanisms[J].Oncol Rep,2016,36(2):909-917.
[28]Park HK,Jo W,Choi HJ,et al.Time-course changes in the ex pression levels of miR-122,-155,and-21 as markers of liver cell damage,inflamation,and regeneration in acetaminophen-induced liver injury in rats[J].Vet Sci,2016,17(1):45-51.
© 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 |