MicroRNA在SMMC-7721肝癌细胞差异表达谱的研究
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摘要
背景和目的
MicroRNA(miRNA)是一类广泛分布在动植物中的高度保守的非编码单链小RNA (nod-coding small RNA),长度大约有19nt～23nt,它的功能是负调控基因表达。目前,随着对miRNA研究的不断深入,发现了其许多未知的功能,包括调节细胞增殖、分化和凋亡,主要是靠调控信号分子的表达实现,这些信号分子包括细胞因子、生长因子、转录因子、促凋亡和抗凋亡因子。通过激活原癌基因的表达以及抑癌基因的突变缺失,从而引起肿瘤的发生发展。因而：miRNA与恶性肿瘤的发生、发展、预后有着密切的关联。
而肝细胞癌(hepatocellular carcinoma, HCC)又是我国很常见的恶性肿瘤,其死亡率在各类肿瘤致死率中排名第二。肝细胞癌的发生、发展是多因素、多步骤的一系列复杂过程。所以,寻找肝细胞癌发生、发展的相关基因,了解肝细胞癌的分子遗传学机制从而为肝细胞癌的诊断、治疗提供理论基础为目前的研究热点。miRNA的研究为肝癌的研究提供了一个新的思路和希望。
但是迄今为止,肝癌的发生,发展的miRNA相关的分子机制仍不是很清楚,进一步探索肝癌的miRNA分子发生机制对提高肝癌的诊断与治疗水平有很重要的意义,因此本研究拟采用miRNA芯片技术筛选肝癌细胞株SMMC-7721和人胚肝细胞株CCC-HEL-1差异表达的miRNAs,建立其miRNA表达谱,筛选感兴趣的miRNA,为以后的研究提供依据。并探讨其与肝癌的发生发展的关系,以期为阐明肝癌的发生的分子机制提供新的依据。
方法
体外培养人肝癌细胞株SMMC-7721和人胚肝细胞株CCC-HEL-1, Trizol法抽取总的RNA后,紫外吸收测定法在260nm和280nm以及变性琼脂糖凝胶电泳鉴定RNA的质量和浓度,用的miRCURYTM Array Power Labeling kit分离并Hy3荧光标记miRNA,将Hy3荧光标记的miRNA进行与miRNA芯片杂交反应,洗片后,利用生物芯片扫描仪扫描芯片,将图像信号转换为数字信号,进行数据分析和统计学处理,筛选有统计学意义的差异miRNA,导入Cluster3.0进行聚类分析,并挑选感兴趣的miRNA应用Real time PCR方法进行验证。
结果
芯片结果显示,以两组细胞株差异表达的miRNAs (P<0.05)以上调或者下调两倍(fold change>2)作为筛选标准。发现SMMC-7721较CCC-HEL-1细胞株差异表达的miRNAs共238个。上调的miRNAs154个,下调的miRNAs有84个。上调4倍(fold change>4)的miRNAs共64个,上调10倍(fold change >10)的miRNAs共24个,由高到低依次为hsa-miR-205,hsa-miR-16,hsa-miR-1 01,hsa-miR-195,hsa-miR-30b,hsa-miR-30e,hsa-miRPlus-E1209,hsa-miR-200a,hsa-m iR-378,hsa-miR-15a,hsa-let-7f,hsa-miR-141,hsa-miR-424,hsa-miR-181a-2*,hsa-miR-24,hsa-miR-30a,hsa-let-7a,hsa-miR-26a,hsa-miR-26a-2*,hsa-miR-130a,hsa-miR-15b, hsa-miR-23a,hsa-miR-27a,hsa-miR-193b,hsa-miRPlus-C1005,其中上调最高的hsa-miR-205上调52倍,下调的miRNAs共84个,下调4倍(fold change>4)的有22个,下调5倍(fold change>5)的有10个,为下调倍数由高到低依次为hsa-miRPlus-E1120, hsa-miRPlus-A1027, hsa-miRPlus-E1012, hsa-miR-608, hsa-miRPlus-F1208, hsa-miRPlus-F1205, hsa-miR-767-3p, hsa-miR-373*, hsa-mi R-769-3p, hsa-miRPlus-E1245,其中的hsa-miRPlus-E1120下调13倍
应用Realtime PCR方法检测let-7f在SMMC-7721-1和CCC-HEL-1-4中表达情况,溶解曲线呈单峰,显示引物的特异性良好,结果显示,与CCC-HEL-1细胞株相比,SMMC-7721细胞株中的let-7f表达上调,为上调2.3倍。
应用Realtime PCR方法检测miR-205在SMMC-7721-1和CCC-HEL-1-4由表达情况,溶解曲线呈单峰,显示引物的特异性良好,结果显示,与CCC-HEL.1细胞株相比,SMMC-7721细胞株中的miR-205表达上调,为上调2.7倍。
结论
1、实验结果提示人肝癌细胞株SMMC-7721与人胚肝细胞株CCC-HEL-1存在差异性的,miRNA。建立了其miRNA差异表达谱。为进一步研究差异miRNA提供依据。
2、经过Realtime PCR验证,miR-205和let-7f在肝癌细胞株中明显上调。为其功能学检测提供依据。
3、hsa-miRPlus-E1120, hsa-miRPlus-A1027, hsa-miRPlus-E1012, hsa-miR-608,等miRNA在肝癌中下调。
Background and objective
MicroRNA (miRNA) are a class of non-coding small RNA (nod-coding small RNA), They are widely distributed in plants and animals with length of about 19nt～23nt single-stranded RNA, its function is a negative regulator of gene expression. Now, with the deepening research of miRNA and many of its un known functions were found, including regulation of cell proliferation, differen tiation and apoptosis, mainly by regulating the expression of signaling molecul es to achieve. These signaling molecules including cytokines, growth factors, tr anscription factor, pro-apoptotic and anti-apoptotic factor. By activating the exp ression of proto-oncogenes and tumor suppressor gene mutation deletion
Which led to the development of tumors. MiRNA are closely related to t he occurrence, development, and prognosis of cancer
HCC (hepatocellular carcinoma, HCC) is a common malignant tumor in C hina, it's mortality rate is second highest in the malignant tumor .The occurre nce and development of HCC is a multi-factor, multi-step complex process. Lo oking for occurrence and development related genes of HCC, in order to un derstand the molecular genetic mechanism of hepatocellular carcinoma so as to HCC diagnosis, treatment and providing a theoretical foundation is the current research focus. The research of miRNA provides a new idea and hope for ca ncer research
But so far, the occurrence of liver cancer and development related to the molecular mechanism of miRNA is still not very clear, and further explore m olecules mechanism of the miRNA in liver cancer will improving the diagnos is and treatment of liver cancer and this is very important, the present study was screening differentially expressed miRNAs between hepatoma cell line SM MC-7721 and human embryo liver cell line CCC-HEL-1 by miRNA microarra y, and to explore the relationship between the occurrence and development of liver cancer.It will clarify the molecular mechanisms of liver cancer.
the present study was screening differentially expressed miRNAs between hepatoma cell line SMMC-7721 and human embryo liver cell line CCC-HEL-1 by miRNA microarray, and to explore the relationship between the occurrenc e and development of liver cancer.It will clarify the molecular mechanisms of liver cancer.
validate differential expression of MicroRNA to explore the relationship be tween differential expression of miRNA and liver cancer by Real-time quantitat ive PCR
Methods
Human hepatoma cell line SMMC-7721 and human embryo liver cell line CC C-HEL-1 were purchased from Chinese Academy of Medical Sciences, Institute of Basic Medical Cell Center of Basic Medicine,Total RNA extraction by Tr izol method, Identify the quality and concentration of RNA by NanoDro p ND-1000 at 260nm and 280nm and denaturing Agarose Gel Electrophoresi s .Isolating and labeling miRNA by Hy3 and Exiqon's miRCURYTM Array Power Labeling kit, the miRNA was hybridized on miRNAarray, Scanning is performed with the Axon GenePix 4000B microarray scanner.The result of miRNA array was verified by real time PCR
Results
By means of up-regulation or down-regulation up to twice (fold change> 2) a s a screening criteria, This study showed that total 238 differential expressed miRNAs were found, including 154 over-expression and 84 low-expression mi RNAs. Four-fold increase (fold change> 4) of the miRNAs were 64,10-fold i ncrease (fold change>10) of the miRNAs were24,ranked as hsa-miR-205, hsa-m iR-16, hsa-miR-101, hsa-miR-195, hsa-miR-30b, hsa-miR-30e,hsa-miRPlus-E1209, hsa-miR-200a,hsa-miR-378,hsa-miR-15a,hsa-let-7f,hsa-miR-141,hsa-miR-424,hsa-m iR-181 a-2*,hsa-miR-24,hsa-miR-30a,hsa-let-7a,hsa-miR-26a,hsa-miR-26a-2*,hsa-mi R-130a,hsa-miR-15b,hsa-miR-23a,hsa-miR-27a,hsa-miR-193b,hsa-miRPlus-C1005, The highest fold change was up to 52 (P< 0.05). A total of 84 of the miRN As down-regulate, there were 22 miRNAs are up to 4 times (fold change> 4) , there were 10 are up to 5 times (fold change> 5), ranked as hsa-miRPl us-E1120, hsa-miRPlus-A1027, hsa-miRPlus-E1012, hsa-miR-608, hsa-miRPlus-F 1208, hsa-miRPlus-F1205, hsa-miR-767-3p, hsa-miR-373*, hsa-miR-769 -3p, hs a-miRPlus-E1245, which hsa-miRPlus-E1120 reduced 13-fold.
Application Realtime PCR to detect the expression of let-7f between S MMC-7721-1 and CCC-HEL-1-4.The dissolution curve has a single peak, This indicating a good specificity of the primers .Compare to CCC-HEL-1 cell line , The expression of let-7f is increase 2.3 times in SMMC-7721 cell line.
Application Realtime PCR to detect the expression of miR-205 between
SMMC-7721-1 and CCC-HEL-1-4.The dissolution curve has a single peak. This indicating a good specificity of the primers .Compare to CCC-HEL-1 cell line. The expression of miR-25 is increase 2.7times in SMMC-7721 cell line
Couclusion
1, The result suggests that there are different miRNA between the human hepatoma cell line SMMC-7721 and human fetal liver cell line CCC-HEL-1 Established between the miRNA expression profiles.
2, After validation of Realtime PCR, miR-205 and let-7f were up-regulatio n in liver cancer cell lines
3, Hsa-miRPlus-E1120, hsa-miRPlus-A1027, hsa-miRPlus-E1012, hsa-miR-6 08, and other miRNA are down-regulation in liver cancer cell line.
MicroRNA(miRNA)是一类广泛分布在动植物中的高度保守的非编码单链小RNA (nod-coding small RNA),长度大约有19nt～23nt,它的功能是负调控基因表达。目前,随着对miRNA研究的不断深入,发现了其许多未知的功能,包括调节细胞增殖、分化和凋亡,主要是靠调控信号分子的表达实现,这些信号分子包括细胞因子、生长因子、转录因子、促凋亡和抗凋亡因子。通过激活原癌基因的表达以及抑癌基因的突变缺失,从而引起肿瘤的发生发展。因而：miRNA与恶性肿瘤的发生、发展、预后有着密切的关联。
而肝细胞癌(hepatocellular carcinoma, HCC)又是我国很常见的恶性肿瘤,其死亡率在各类肿瘤致死率中排名第二。肝细胞癌的发生、发展是多因素、多步骤的一系列复杂过程。所以,寻找肝细胞癌发生、发展的相关基因,了解肝细胞癌的分子遗传学机制从而为肝细胞癌的诊断、治疗提供理论基础为目前的研究热点。miRNA的研究为肝癌的研究提供了一个新的思路和希望。
但是迄今为止,肝癌的发生,发展的miRNA相关的分子机制仍不是很清楚,进一步探索肝癌的miRNA分子发生机制对提高肝癌的诊断与治疗水平有很重要的意义,因此本研究拟采用miRNA芯片技术筛选肝癌细胞株SMMC-7721和人胚肝细胞株CCC-HEL-1差异表达的miRNAs,建立其miRNA表达谱,筛选感兴趣的miRNA,为以后的研究提供依据。并探讨其与肝癌的发生发展的关系,以期为阐明肝癌的发生的分子机制提供新的依据。
方法
体外培养人肝癌细胞株SMMC-7721和人胚肝细胞株CCC-HEL-1, Trizol法抽取总的RNA后,紫外吸收测定法在260nm和280nm以及变性琼脂糖凝胶电泳鉴定RNA的质量和浓度,用的miRCURYTM Array Power Labeling kit分离并Hy3荧光标记miRNA,将Hy3荧光标记的miRNA进行与miRNA芯片杂交反应,洗片后,利用生物芯片扫描仪扫描芯片,将图像信号转换为数字信号,进行数据分析和统计学处理,筛选有统计学意义的差异miRNA,导入Cluster3.0进行聚类分析,并挑选感兴趣的miRNA应用Real time PCR方法进行验证。
结果
芯片结果显示,以两组细胞株差异表达的miRNAs (P<0.05)以上调或者下调两倍(fold change>2)作为筛选标准。发现SMMC-7721较CCC-HEL-1细胞株差异表达的miRNAs共238个。上调的miRNAs154个,下调的miRNAs有84个。上调4倍(fold change>4)的miRNAs共64个,上调10倍(fold change >10)的miRNAs共24个,由高到低依次为hsa-miR-205,hsa-miR-16,hsa-miR-1 01,hsa-miR-195,hsa-miR-30b,hsa-miR-30e,hsa-miRPlus-E1209,hsa-miR-200a,hsa-m iR-378,hsa-miR-15a,hsa-let-7f,hsa-miR-141,hsa-miR-424,hsa-miR-181a-2*,hsa-miR-24,hsa-miR-30a,hsa-let-7a,hsa-miR-26a,hsa-miR-26a-2*,hsa-miR-130a,hsa-miR-15b, hsa-miR-23a,hsa-miR-27a,hsa-miR-193b,hsa-miRPlus-C1005,其中上调最高的hsa-miR-205上调52倍,下调的miRNAs共84个,下调4倍(fold change>4)的有22个,下调5倍(fold change>5)的有10个,为下调倍数由高到低依次为hsa-miRPlus-E1120, hsa-miRPlus-A1027, hsa-miRPlus-E1012, hsa-miR-608, hsa-miRPlus-F1208, hsa-miRPlus-F1205, hsa-miR-767-3p, hsa-miR-373*, hsa-mi R-769-3p, hsa-miRPlus-E1245,其中的hsa-miRPlus-E1120下调13倍
应用Realtime PCR方法检测let-7f在SMMC-7721-1和CCC-HEL-1-4中表达情况,溶解曲线呈单峰,显示引物的特异性良好,结果显示,与CCC-HEL-1细胞株相比,SMMC-7721细胞株中的let-7f表达上调,为上调2.3倍。
应用Realtime PCR方法检测miR-205在SMMC-7721-1和CCC-HEL-1-4由表达情况,溶解曲线呈单峰,显示引物的特异性良好,结果显示,与CCC-HEL.1细胞株相比,SMMC-7721细胞株中的miR-205表达上调,为上调2.7倍。
结论
1、实验结果提示人肝癌细胞株SMMC-7721与人胚肝细胞株CCC-HEL-1存在差异性的,miRNA。建立了其miRNA差异表达谱。为进一步研究差异miRNA提供依据。
2、经过Realtime PCR验证,miR-205和let-7f在肝癌细胞株中明显上调。为其功能学检测提供依据。
3、hsa-miRPlus-E1120, hsa-miRPlus-A1027, hsa-miRPlus-E1012, hsa-miR-608,等miRNA在肝癌中下调。
Background and objective
MicroRNA (miRNA) are a class of non-coding small RNA (nod-coding small RNA), They are widely distributed in plants and animals with length of about 19nt～23nt single-stranded RNA, its function is a negative regulator of gene expression. Now, with the deepening research of miRNA and many of its un known functions were found, including regulation of cell proliferation, differen tiation and apoptosis, mainly by regulating the expression of signaling molecul es to achieve. These signaling molecules including cytokines, growth factors, tr anscription factor, pro-apoptotic and anti-apoptotic factor. By activating the exp ression of proto-oncogenes and tumor suppressor gene mutation deletion
Which led to the development of tumors. MiRNA are closely related to t he occurrence, development, and prognosis of cancer
HCC (hepatocellular carcinoma, HCC) is a common malignant tumor in C hina, it's mortality rate is second highest in the malignant tumor .The occurre nce and development of HCC is a multi-factor, multi-step complex process. Lo oking for occurrence and development related genes of HCC, in order to un derstand the molecular genetic mechanism of hepatocellular carcinoma so as to HCC diagnosis, treatment and providing a theoretical foundation is the current research focus. The research of miRNA provides a new idea and hope for ca ncer research
But so far, the occurrence of liver cancer and development related to the molecular mechanism of miRNA is still not very clear, and further explore m olecules mechanism of the miRNA in liver cancer will improving the diagnos is and treatment of liver cancer and this is very important, the present study was screening differentially expressed miRNAs between hepatoma cell line SM MC-7721 and human embryo liver cell line CCC-HEL-1 by miRNA microarra y, and to explore the relationship between the occurrence and development of liver cancer.It will clarify the molecular mechanisms of liver cancer.
the present study was screening differentially expressed miRNAs between hepatoma cell line SMMC-7721 and human embryo liver cell line CCC-HEL-1 by miRNA microarray, and to explore the relationship between the occurrenc e and development of liver cancer.It will clarify the molecular mechanisms of liver cancer.
validate differential expression of MicroRNA to explore the relationship be tween differential expression of miRNA and liver cancer by Real-time quantitat ive PCR
Methods
Human hepatoma cell line SMMC-7721 and human embryo liver cell line CC C-HEL-1 were purchased from Chinese Academy of Medical Sciences, Institute of Basic Medical Cell Center of Basic Medicine,Total RNA extraction by Tr izol method, Identify the quality and concentration of RNA by NanoDro p ND-1000 at 260nm and 280nm and denaturing Agarose Gel Electrophoresi s .Isolating and labeling miRNA by Hy3 and Exiqon's miRCURYTM Array Power Labeling kit, the miRNA was hybridized on miRNAarray, Scanning is performed with the Axon GenePix 4000B microarray scanner.The result of miRNA array was verified by real time PCR
Results
By means of up-regulation or down-regulation up to twice (fold change> 2) a s a screening criteria, This study showed that total 238 differential expressed miRNAs were found, including 154 over-expression and 84 low-expression mi RNAs. Four-fold increase (fold change> 4) of the miRNAs were 64,10-fold i ncrease (fold change>10) of the miRNAs were24,ranked as hsa-miR-205, hsa-m iR-16, hsa-miR-101, hsa-miR-195, hsa-miR-30b, hsa-miR-30e,hsa-miRPlus-E1209, hsa-miR-200a,hsa-miR-378,hsa-miR-15a,hsa-let-7f,hsa-miR-141,hsa-miR-424,hsa-m iR-181 a-2*,hsa-miR-24,hsa-miR-30a,hsa-let-7a,hsa-miR-26a,hsa-miR-26a-2*,hsa-mi R-130a,hsa-miR-15b,hsa-miR-23a,hsa-miR-27a,hsa-miR-193b,hsa-miRPlus-C1005, The highest fold change was up to 52 (P< 0.05). A total of 84 of the miRN As down-regulate, there were 22 miRNAs are up to 4 times (fold change> 4) , there were 10 are up to 5 times (fold change> 5), ranked as hsa-miRPl us-E1120, hsa-miRPlus-A1027, hsa-miRPlus-E1012, hsa-miR-608, hsa-miRPlus-F 1208, hsa-miRPlus-F1205, hsa-miR-767-3p, hsa-miR-373*, hsa-miR-769 -3p, hs a-miRPlus-E1245, which hsa-miRPlus-E1120 reduced 13-fold.
Application Realtime PCR to detect the expression of let-7f between S MMC-7721-1 and CCC-HEL-1-4.The dissolution curve has a single peak, This indicating a good specificity of the primers .Compare to CCC-HEL-1 cell line , The expression of let-7f is increase 2.3 times in SMMC-7721 cell line.
Application Realtime PCR to detect the expression of miR-205 between
SMMC-7721-1 and CCC-HEL-1-4.The dissolution curve has a single peak. This indicating a good specificity of the primers .Compare to CCC-HEL-1 cell line. The expression of miR-25 is increase 2.7times in SMMC-7721 cell line
Couclusion
1, The result suggests that there are different miRNA between the human hepatoma cell line SMMC-7721 and human fetal liver cell line CCC-HEL-1 Established between the miRNA expression profiles.
2, After validation of Realtime PCR, miR-205 and let-7f were up-regulatio n in liver cancer cell lines
3, Hsa-miRPlus-E1120, hsa-miRPlus-A1027, hsa-miRPlus-E1012, hsa-miR-6 08, and other miRNA are down-regulation in liver cancer cell line.
引文
[1]BartelDP.MicroRNAs:genomics,biogenesis,mechanism,andfunction[J].Cell,2004,116(2):281-297
[2]Lee RC, Feinbaum RL, AmbiosV. The C. elegans gene heterochronic gene lin-4 encodes small RNAs antisense complementarity to lin-14[J].Cell,1993,75(5):843-854
[3]Reinhart BJ, Slack FJ, Basson M,et al. The 21-nucleotide let-7 RNA regulates developmental timing in Caenorhabditis elegans[J].Nature,403(6772):901-6
[4]Slack FJ,basson M,Liu Z,et al,The lin-41 RBCC gene acts in the C.elegans heterochronic pathway between the let-7 regulatory RNA and the LIN-29 transcription factor[J],Mol Cell,2000,5(4):659-669
[5]Berezikov E,Guryev v,van de Belt J,et al.Phylogenetic shadowing and computational identification of human microRNA genes [J].Cell,2005,120(1):21-24
[6]Grad Y,Aach J,Hayes GD,et al.Computational and experimental identification of C.elegans microRNAs[J].Mol Cell,2003,11(5):1253-1263
[7]Lau NC,Lim LP,Weinstein EG,et al.An abundant class of tiny RNAs with probable regulatory roles in Caenorhabditis elegans[J].Science,2001,294(5543):858-862
[8][8]Rodriguez A,Griffiths-Jones S,Ashurst JL,et al.Identification of mammalian microRNA host genes and transcription units[J].Genome Res,2004,14(10A):1902-1910
[9]Khvorova A,Reynolds A,Jayasena SD.Functional siRNAs and miRNAs exhibit strand bias[J].Cell,2003,115(2):209-216
[10]Hutvagner G,Zamore PD.A microRNA in a multiple-turnover RNAi enzyme complex[J]. Science,2002,297(5589):2056-2060
[11]Zeng Y,Yi R,Cullen BR.MicroRNAs and small interfering RNAs can inhibit mRNA expression by similar mechanisms [J].Proc Natl Acad Sci USA,2003,100(17):9779-9784
[12]Lai EC.Micro RNAs are complementary to 3'UTRsequence motifs tha mediate negative post-transcriptional regulation[J].Nat Genet,2002,30(4):363-364
[13]戚鹏,韩金祥,鲁艳芹.疱疹病毒编码的miRNAs的研究进展.中国生物工程杂志,2006,26：69～74
[14]Johnson CD, Esquela-Kerscher A, Stefani G ,et al.The let-7 microRNA represses cell proliferation pathways in human cells[J]. Cancer Res.2007 67(16):7713-22
[15]Hanlon K, Rudin CE, Harries LW. Investigating the targets of MIR-15a and MIR-16-1 in patients with chronic lymphocytic leukemia (CLL)[J].PloS One.2009 Sep 25:4(9)e7169
[16]Chan JA, Krichevsky AM, Kosik KS. MicroRNA-21 is an antiapoptotic factor in human glioblastoma cells[J].Cancer Res.2005;65(14):6029-33
[17]Takagi T, Iio A, Nakagawa Y, Naoe T,et al. Decreased expression of microRNA-143 and-145 in human gastric cancers[J].Oncology,2009,77(1):12-21
[18]Kluiver J, Haralambieva E, de Jong D,et al.Lack of BIC and microRNA miR-155 expression in primary cases of Burkitt lymphoma[J].Genes Chromosomes Cancer.2006,45(2):147-53
[19]Pineau P, Volinia S, McJunkin K,et al. miR-221 overexpression contributes to liver tumorigenesis[J].Proc Natl Acad Sci U S A.2010,107(1):264-9
[20]Yamamoto Y, Kosaka N, Tanaka M,Biomarkers.et al. MicroRNA-500 as a potential diagnostic marker for hepatocellular carcinoma[J].2009,14(7):529-38
[21]Wang Y, Lee AT, Ma JZ, et al. Profiling microRNA expression in hepatocellular carcinoma reveals microRNA-224 up-regulation and apoptosis inhibitor-5 as a microRNA-224-specific target[J]. J Biol Chem.2008;283(19):13205-15
[22]Guo Y, Chen Y, Ito H,et al.Identification and characterization of lin-28 homolog B (LIN28B) in human hepatocellular carcinoma[J]. Gene.2006,384:51-61
[23][23] Huang YS, Dai Y, Yu XF,et al Microarray analysis of microRNA expression in hepatocellular carcinoma and non-tumorous tissues without viral hepatitis.J Gastroenterol Hepatol[J].2008,23(1):87-94
[24]Thorgeirsson SS, Lee JS, Grisham JW.Functional genomics of hepatocellular carcinoma[J]. Hepatology.200643(2 Suppl 1):S145-50
[25]Huang S, He X, Ding J,Upregulation of miR-23a approximately 27a approximately 24 decreases transforming growth factor-beta-induced tumor-suppressive activities in human hepatocellular carcinoma cells[J].Int J Cancer.2008,123(4):972-8
[26]Fornari F, Gramantieri L, Ferracin M, et al.MiR-221 controls CDKN1C/p57 and CDKN1B/ p27 expression in human hepatocellular carcinoma[J].Oncogene.2008,27(43):5651-61
[27]Datta J, Kutay H, Nasser MW,et al.Methylation mediated silencing ofMicroRNA-1 gene and its role in hepatocellular carcinogenesis[J].Cancer Res.2008,68(13):5049-58
[28]Li W, Xie L, He X,et al Diagnostic and prognostic implications of microRNAs in human hepatocellular carcinoma.Int J Cancer[J].2008,123(7):1616-22
[29]Arbuthnot P, Thompson LJ.Harnessing the RNA interference pathway to advance treatment and prevention of hepatocellular carcinoma[J]. World J Gastroenterol.2008,14(11):1670-81
[30]Sun BS, Dong QZ, Ye QH, et al,Lentiviral-mediated miRNA against osteopontin suppresses tumor growth and metastasis of human hepatocellular carcinoma[J]. Hepatology,48(6): 1834-42
[31]Chen C,Ridzon DA,Broomer AJ,et al.Real-time quantification of microRNAs by stem-loop RT-PCR[J].Nucleic Acids Res,2005,33(20):el79
[32]谢纳,生物芯片分析[M].北京：科学出版社,2003
[33]马立人,蒋中华主编,生物芯片[M].北京：化学工业出版社,2000
[34]Lagos-Quintana M, Rauhut R, Lendeckel W,et al.Identification of novel genes coding for small expressed RNAs[J].Tuschl T Science.2001.294:853-858
[35]Schultz J, Lorenz P, Gross G, et al.MicroRNA let-7b targets important cell cycle molecules in malignant melanoma cells and interferes with anchorage-independent growth[J]. Cell Res. 2008.18(5):549-57
[36]Slack F.let-7 microRNA reduces tumor growth.Cell Cycle.2009,8(12):1823
[37]Shi G, Perle MA, Mittal K,et al.Let-7 repression leads to HMGA2 overexpression in uterine leiomyosarcoma[J].J Cell Mol Med.2009,(9B):3898-905
[38]Ren Y, Kang CS, Yuan XB, et al.Co-delivery of as-miR-21 and 5-FU by poly(amidoamine) dendrimer attenuates human glioma cell growth in vitro[J].J Biomater Sci Polym Ed. 2010;21(3):303-14
[39]Volinia S, Calin GA, Liu CG, et al.A microRNA expression signature of human solid tumors defines cancer gene targets.Proc Natl Acad Sci U S A.2006,103(7):2257-61
[40]Meng F, Henson R, Wehbe-Janek H, et al. MicroRNA-21 regulates expression of the PTEN tumor suppressor gene in human hepatocellular cancer[J].Gastroenterology.2007133(2): 647-58
[41]Volinia S,Calin GA,Liu C G, et al. A microRNA expression signature of human solid tumors defines cancer gene targets[J],2006,103:2257～2261
[42]Akao Y,Nakagawa Y,Naoe T. Let-7 microRNA functions as a potential growth suppressor in human colon cancer cells[J]. Biol Pharm Bull,2006,29:903～906
[43]SiM L, Zhu S,Wu H, et al. MiR-21-mediated tumor growth[J].2007,26:2799～2803
[44]Kutay H, Bai S, Datta J, et al.Downregulation of miR-122 in the rodent and human hepatocellular carcinomas[J]. J Cell Biochem.2006,99(3):671-8
[45]Pogribny IP, Bagnyukova TV, Tryndyak VP, et al. Gene expression profiling reveals underlying molecular mechanisms of the early stages of tamoxifen-induced rat hepatocarcinogenesis[J].2007,225(1):61-9
[46]Weber F, Teresi RE, Broelsch CE,et al. A limited set of human MicroRNA is deregulated in follicular thyroid carcinoma[J].J Clin Endocrinol Metab.2006,91(9):3584-91
[47][47]Ladeiro Y, Couchy G, Balabaud C,et al.MicroRNA profiling in hepatocellular tumors is associated with clinical features and oncogene/tumor suppressor gene mutations. Hepatology.2008,47(6):1955-63
[48]Murakami Y, Yasuda T, Saigo K,et al.Comprehensive analysis of microRNA expression patterns in hepatocellular carcinoma and non-tumorous tissues[J].Oncogene.2006 ,25(17):2537-45
[49]Gramantieri L, Fornari F, Callegari E,et al.MicroRNA involvement in hepatocellular carcinoma[J].J Cell Mol Med.2008,12(6A):2189-204
[50]Danit Lebanony, Hila Benjamin, Shlomit Gilad Diagnostic Assay Based on hsa-miR-205 Expression Distinguishes Squamous From Nonsquamous Non-Small-Cell Lung Carcinoma[J].Journal of Clinical Oncology, Vol 27, No 12 (April 20),2009:pp.2030-2037
[51]孙小丽,林仲秋,吴维光.上调microRNA-205表达对HeLa细胞生物学特性的影响 中山大学学报(医学科学版),2009/30/02
[52]Geoffrey Childs*, Melissa Fazzari, Low-Level Expression of MicroRNAs let-7d and miR-205 Are Prognostic Markers of Head and Neck Squamous Cell Carcinoma American Journal of Pathology[J].2009; 174:736-745
[53]lorio MV, Casalini P, microRNA-205 regulates HER3 in human breast cancer[J]. Cancer Res.2009 Mar 15;69(6):2195-200
[54]Wu H, Mo YY. Targeting miR-205 in breast cancer Expert Opin Ther Targets[J].2009 Oct 19
[1]Nishida N.Impact of hepatitis virus and aging on DNA methylation in human hepatocarcinogenesis[J].Histol Histopathol.2010,25(5):647-54
[2]Yuan A, Li Z, Li X,et al, The mitogenic effectors of isoproterenol in human hepatocellular carcinoma cells[J].Oncol Rep.2010,23(1):151-7
[3]Bishayee A, Politis T, Darvesh AS.Resveratrol in the chemoprevention and treatment of hepatocellularcarcinoma[J].Cancer Treat Rev.2010,36(1):43-53
[4]Arbuthnot P, Thompson LJ.Harnessing the RNA interference pathway to advance treatment and prevention of hepatocellular carcinoma[J].World J Gastroenterol.2008,14(11):1670-81
[5]Chen Y, Cheng G, Mahato RI.RNAi for treating hepatitis B viral infection[J].Pharm Res. 2008,25(1):72-86
[6]Roessler S, Budhu A, Wang XW.Future of molecular profiling of human hepatocellular carcinoma.Future Oncol[J].2007,3(4):429-39
[7]Lee RC, Feinbaum RL, AmbiosV. The C. elegans gene heterochronic gene lin-4 encodes small RNAs antisense complementarity to lin-14[J].Cell,1993,75(5):843-854
[8]Lagos-Quintana M, Rauhut R, Lendeckel W, et al. Identification of novel genes coding for small expressed RNAs[J]. Science,2001,294:853～858
[9]Lau NC,Lim LP,Weinstein EG,et al.An abundant class of tiny RNAs with probable regulatory roles in Caenorhabditis elegans[J].Science,2001,294(5543):858-862
[10]Rodriguez A,Griffiths-Jones S,Ashurst JL,et al.Identification of mammalian microRNA host genes and transcription units [J].Genome Res,2004,14(10A):1902-1910
[11]Khvorova A,Reynolds A,Jayasena SD.Functional siRNAs and miRNAs exhibit strand bias[J].Cell,2003,115(2):209-216
[12]He L, Hannon GJ. MicroRNAs:small RNAs with a big role in gene regulation[J]. NatRev Genet,2004,5:522～531
[13]Gregory RI, Shiekhattar R. MicroRNA biogenesis and cancer [J]. Cancer Res,2005 ,65(9):3509-12
[14]Hutvagner G,Zamore PD.A microRNA in a multiple-turnover RNAi enzyme complex[J].Science,2002,297(5589):2056-2060
[15]Zeng Y,Yi R,Cullen BR.MicroRNAs and small interfering RNAs can inhibit mRNA expression by similar mechanisms [J].Proc Natl Acad Sci USA,2003,100(17):9779-9784
[16]Lai EC.Micro RNAs are complementary to 3'UTRsequence motifs tha mediate negative post-transcriptional regulation[J].Nat Genet,2002,30(4):363-364
[17]Davidson-Moncada J, Papavasiliou FN, Tam W.MicroRNAs of the immune system: roles in inflammation and cancer[J].Ann N Y Acad Sci.2010,1183:183-94
[18]Zhang H, Li Y, Lai M.The microRNA network and tumor metastasis.Oncogene.2010 29(7):937-48.
[19]Davidson-Moncada J, Papavasiliou FN, Tam W.MicroRNAs of the immune system:roles in inflammation and cancer[J]. Ann N Y Acad Sci.2010,1183:183-94. Review
[20]Shenouda SK, Alahari SK.MicroRNA function in cancer: oncogene or a tumor suppressor[J]? Cancer Metastasis Rev.2009,28(3-4):369-78
[21]Trang P, Weidhaas JB, Slack FJ.MicroRNAs as potential cancer therapeutics[J].Oncogene. 2008, Suppl 2:S52-7
[22]戚鹏,韩金祥,鲁艳芹.疱疹病毒编码的miRNAs的研究进展.中国生物工程杂志,2006,26：69～74
[23]Davalos V, Esteller M.MicroRNAs and cancer epigenetics:a acrorevolution[J].Curr Opin Oncol.2010,22(1):35-45
[24]Taft RJ, Pang KC, Mercer TR, et al, Non-coding RNAs: regulators of disease[J].J Pathol. 2010,220(2):126-39
[25]Baranwal S, Alahari SK.miRNA control of tumor cell invasion and metastasis[J].Int J Cancer.2010,126(6):1283-90
[26]Johnson CD, Esquela-Kerscher A, Stefani G ,et al.The let-7 microRNA represses cell proliferation pathways in human cells[J]. Cancer Res.2007 67(16):7713-22
[27]Chan JA, Krichevsky AM, Kosik KS. MicroRNA-21 is an antiapoptotic factor in human glioblastoma cells[J].Cancer Res.2005;65(14):6029-33
[28]Takagi T, Iio A, Nakagawa Y, Naoe T,et al. Decreased expression of microRNA-143 and-145 in human gastric cancers[J].Oncology,2009,77(1):12-21.
[29]Ren Y, Kang CS, Yuan XB, et al.Co-delivery of as-miR-21 and 5-FU by poly(amidoamine) dendrimer attenuates human glioma cell growth in vitro[J].J Biomater Sci Polym Ed. 2010;21(3):303-14.
[30]Kluiver J, Haralambieva E, de Jong D,et al.Lack of BIC and microRNA miR-155 expression in primary cases of Burkitt lymphoma[J].Genes Chromosomes Cancer.2006,45(2):147-53.
[31]Navon R, Wang H, Steinfeld I,et al. Novel rank-based statistical methods reveal microRNAs with differential expression in multiple cancer types[J].PLoS One.2009,4(11):8003
[32]Calin GA.MicroRNAs and cancer:what we know and what we still have to learn[J].Genome Med.2009,1(8):78
[33]谢纳,生物芯片分析[M].北京：科学出版社,2003
[34]马立人,蒋中华主编,生物芯片[M].北京：化学工业出版社,2000
[35]Pineau P, Volinia S, McJunkin K,et al. miR-221 overexpression contributes to liver tumorigenesis[J].Proc Natl Acad Sci U S A.2010,107(1):264-9
[36]Yamamoto Y, Kosaka N, Tanaka M,Biomarkers.et al. MicroRNA-500 as a potential diagnostic marker for hepatocellular carcinoma[J].2009,14(7):529-38
[37]Wang Y, Lee AT, Ma JZ, et al. Profiling microRNA expression in hepatocellular carcinoma reveals microRNA-224 up-regulation and apoptosis inhibitor-5 as a microRNA-224-specific target[J]. J Biol Chem.2008;283(19):13205-15
[38]Guo Y, Chen Y, Ito H,et al.Identification and characterization of lin-28 homolog B (LIN28B) in human hepatocellular carcinoma[J]. Gene.2006,384:51-61
[39]Huang YS, Dai Y, Yu XF,et al Microarray analysis of microRNA expression in hepatocellular carcinoma and non-tumorous tissues without viral hepatitis[J].J Gastroenterol Hepatol.2008,23(1):87-94
[40]Thorgeirsson SS, Lee JS, Grisham JW.Functional genomics of hepatocellular carcinoma[J]. Hepatology.200643(2 Suppl 1):S145-50
[41]Huang S, He X, Ding J,Upregulation of miR-23a approximately 27a approximately 24 decreases transforming growth factor-beta-induced tumor-suppressive activities in human hepatocellular carcinoma cells[J].Int J Cancer.2008,123(4):972-8
[42]Fornari F, Gramantieri L, Ferracin M, et al.MiR-221 controls CDKN1C/p57 and CDKN1B/p27 expression in human hepatocellular carcinoma[J]. Oncogene.2008,27(43) :5651-61
[43]Datta J, Kutay H, Nasser MW,et al.Methylation mediated silencing ofMicroRNA-1 gene and its role in hepatocellular carcinogenesis[J].Cancer Res.2008,68(13):5049-58
[44]Li W, Xie L, He X,et al Diagnostic and prognostic implications of microRNAs in human hepatocellular carcinoma.Int J Cancer[J].2008,123(7):1616-22
[45]Arbuthnot P, Thompson LJ.Harnessing the RNA interference pathway to advance treatment and prevention of hepatocellular carcinoma[J]. World J Gastroenterol.2008,14(11):1670-81
[46]Sun BS, Dong QZ, Ye QH, et al,Lentiviral-mediated miRNA against osteopontin suppresses tumor growth and metastasis of human hepatocellular carcinoma[J]. Hepatology,48(6): 1834-42
[47]Pineau P, Volinia S, McJunkin K,et al. miR-221 overexpression contributes to liver tumorigenesis[J].Proc Natl Acad Sci U S A.2010,107(1):264-9
[48]Yamamoto Y, Kosaka N, Tanaka M,Biomarkers.et al. MicroRNA-500 as a potential diagnostic marker for hepatocellular carcinoma[J].2009,14(7):529-38
[49]Murakami Y, Yasuda T, Saigo K,et al. Comprehensive analysis of microRNA expression patterns in hepatocellular carcinoma and non-tumorous tissues[J]. Oncogene.2006,25(17): 2537-45
[50]Wang Y, Lee AT, Ma JZ, et al. Profiling microRNA expression in hepatocellular carcinoma reveals microRNA-224 up-regulation and apoptosis inhibitor-5 as a microRNA-224-specific target[J]. J Biol Chem.2008;283(19):13205-15
[51]Kutay H, Bai S, Datta J, et al.Downregulation of miR-122 in the rodent and human hepatocellular carcinomas[J]. J Cell Biochem.2006,99(3):671-8
[52]Meng F, Henson R, Wehbe-Janek H, et al. MicroRNA-21 regulates expression of the PTEN tumor suppressor gene in human hepatocellular cancer[J].Gastroenterology.2007133(2): 647-58
[53]Gramantieri L, Ferracin M, Fornari F,et al.Cyclin G1 is a target of miR-122a, a microRNA frequently down-regulated in human hepatocellular carcinoma[J]. Cancer Res. 2007.67(13):6092-9
[54]Wu X, Wu S, Tong L,et al. miR-122 affects the viability and apoptosis of hepatocellular carcinoma cells[J]. Scand J Gastroenterol.2009;44(11):1332-9
[55]Arbuthnot P, Thompson LJ.Harnessing the RNA interference pathway to advance treatment and prevention of hepatocellular carcinoma[J].World J Gastroenterol.2008,14(11):1670-81
[56]Chen Y, Cheng G, Mahato RI.RNAi for treating hepatitis B viral infection[J].Pharm Res. 2008,25(1):72-86
[57]Ji J, Shi J, Budhu A,et al. MicroRNA expression, survival, and response to interferon in liver cancer[J].N Engl J Med.2009.361(15):1437-47
[58]Budhu A, Jia HL, Forgues M, et al. Identification of metastasis-related microRNAs in hepatocellular carcinoma[J].Hepatology.2008,47(3):897-907
[59]Olson P, Lu J, Zhang H,et al. MicroRNA dynamics in the stages of tumorigenesis correlate with hallmark capabilities of cancer[J]. Genes Dev.2009,23(18):2152-65.
[60]Volinia S,Calin GA,Liu C G, et al. A microRNA expression signature of human solid tumors defines cancer gene targets[J].2006,103:2257～2261
[61]Akao Y,Nakagawa Y,Naoe T. Let-7 microRNA functions as a potential growth suppressor in human colon cancer cells[J]. Biol Pharm Bull,2006,29:903～906
[62]SiM L, Zhu S,Wu H, et al. MiR-21-mediated tumor growth[J].2007,26:2799～2803
[63]Kota J, Chivukula RR, O'Donnell KA,et al. Therapeutic microRNA delivery suppresses tumorigenesis in a murine liver cancer model[J]. Cell.2009.137(6):1005-17.
[2]Lee RC, Feinbaum RL, AmbiosV. The C. elegans gene heterochronic gene lin-4 encodes small RNAs antisense complementarity to lin-14[J].Cell,1993,75(5):843-854
[3]Reinhart BJ, Slack FJ, Basson M,et al. The 21-nucleotide let-7 RNA regulates developmental timing in Caenorhabditis elegans[J].Nature,403(6772):901-6
[4]Slack FJ,basson M,Liu Z,et al,The lin-41 RBCC gene acts in the C.elegans heterochronic pathway between the let-7 regulatory RNA and the LIN-29 transcription factor[J],Mol Cell,2000,5(4):659-669
[5]Berezikov E,Guryev v,van de Belt J,et al.Phylogenetic shadowing and computational identification of human microRNA genes [J].Cell,2005,120(1):21-24
[6]Grad Y,Aach J,Hayes GD,et al.Computational and experimental identification of C.elegans microRNAs[J].Mol Cell,2003,11(5):1253-1263
[7]Lau NC,Lim LP,Weinstein EG,et al.An abundant class of tiny RNAs with probable regulatory roles in Caenorhabditis elegans[J].Science,2001,294(5543):858-862
[8][8]Rodriguez A,Griffiths-Jones S,Ashurst JL,et al.Identification of mammalian microRNA host genes and transcription units[J].Genome Res,2004,14(10A):1902-1910
[9]Khvorova A,Reynolds A,Jayasena SD.Functional siRNAs and miRNAs exhibit strand bias[J].Cell,2003,115(2):209-216
[10]Hutvagner G,Zamore PD.A microRNA in a multiple-turnover RNAi enzyme complex[J]. Science,2002,297(5589):2056-2060
[11]Zeng Y,Yi R,Cullen BR.MicroRNAs and small interfering RNAs can inhibit mRNA expression by similar mechanisms [J].Proc Natl Acad Sci USA,2003,100(17):9779-9784
[12]Lai EC.Micro RNAs are complementary to 3'UTRsequence motifs tha mediate negative post-transcriptional regulation[J].Nat Genet,2002,30(4):363-364
[13]戚鹏,韩金祥,鲁艳芹.疱疹病毒编码的miRNAs的研究进展.中国生物工程杂志,2006,26：69～74
[14]Johnson CD, Esquela-Kerscher A, Stefani G ,et al.The let-7 microRNA represses cell proliferation pathways in human cells[J]. Cancer Res.2007 67(16):7713-22
[15]Hanlon K, Rudin CE, Harries LW. Investigating the targets of MIR-15a and MIR-16-1 in patients with chronic lymphocytic leukemia (CLL)[J].PloS One.2009 Sep 25:4(9)e7169
[16]Chan JA, Krichevsky AM, Kosik KS. MicroRNA-21 is an antiapoptotic factor in human glioblastoma cells[J].Cancer Res.2005;65(14):6029-33
[17]Takagi T, Iio A, Nakagawa Y, Naoe T,et al. Decreased expression of microRNA-143 and-145 in human gastric cancers[J].Oncology,2009,77(1):12-21
[18]Kluiver J, Haralambieva E, de Jong D,et al.Lack of BIC and microRNA miR-155 expression in primary cases of Burkitt lymphoma[J].Genes Chromosomes Cancer.2006,45(2):147-53
[19]Pineau P, Volinia S, McJunkin K,et al. miR-221 overexpression contributes to liver tumorigenesis[J].Proc Natl Acad Sci U S A.2010,107(1):264-9
[20]Yamamoto Y, Kosaka N, Tanaka M,Biomarkers.et al. MicroRNA-500 as a potential diagnostic marker for hepatocellular carcinoma[J].2009,14(7):529-38
[21]Wang Y, Lee AT, Ma JZ, et al. Profiling microRNA expression in hepatocellular carcinoma reveals microRNA-224 up-regulation and apoptosis inhibitor-5 as a microRNA-224-specific target[J]. J Biol Chem.2008;283(19):13205-15
[22]Guo Y, Chen Y, Ito H,et al.Identification and characterization of lin-28 homolog B (LIN28B) in human hepatocellular carcinoma[J]. Gene.2006,384:51-61
[23][23] Huang YS, Dai Y, Yu XF,et al Microarray analysis of microRNA expression in hepatocellular carcinoma and non-tumorous tissues without viral hepatitis.J Gastroenterol Hepatol[J].2008,23(1):87-94
[24]Thorgeirsson SS, Lee JS, Grisham JW.Functional genomics of hepatocellular carcinoma[J]. Hepatology.200643(2 Suppl 1):S145-50
[25]Huang S, He X, Ding J,Upregulation of miR-23a approximately 27a approximately 24 decreases transforming growth factor-beta-induced tumor-suppressive activities in human hepatocellular carcinoma cells[J].Int J Cancer.2008,123(4):972-8
[26]Fornari F, Gramantieri L, Ferracin M, et al.MiR-221 controls CDKN1C/p57 and CDKN1B/ p27 expression in human hepatocellular carcinoma[J].Oncogene.2008,27(43):5651-61
[27]Datta J, Kutay H, Nasser MW,et al.Methylation mediated silencing ofMicroRNA-1 gene and its role in hepatocellular carcinogenesis[J].Cancer Res.2008,68(13):5049-58
[28]Li W, Xie L, He X,et al Diagnostic and prognostic implications of microRNAs in human hepatocellular carcinoma.Int J Cancer[J].2008,123(7):1616-22
[29]Arbuthnot P, Thompson LJ.Harnessing the RNA interference pathway to advance treatment and prevention of hepatocellular carcinoma[J]. World J Gastroenterol.2008,14(11):1670-81
[30]Sun BS, Dong QZ, Ye QH, et al,Lentiviral-mediated miRNA against osteopontin suppresses tumor growth and metastasis of human hepatocellular carcinoma[J]. Hepatology,48(6): 1834-42
[31]Chen C,Ridzon DA,Broomer AJ,et al.Real-time quantification of microRNAs by stem-loop RT-PCR[J].Nucleic Acids Res,2005,33(20):el79
[32]谢纳,生物芯片分析[M].北京：科学出版社,2003
[33]马立人,蒋中华主编,生物芯片[M].北京：化学工业出版社,2000
[34]Lagos-Quintana M, Rauhut R, Lendeckel W,et al.Identification of novel genes coding for small expressed RNAs[J].Tuschl T Science.2001.294:853-858
[35]Schultz J, Lorenz P, Gross G, et al.MicroRNA let-7b targets important cell cycle molecules in malignant melanoma cells and interferes with anchorage-independent growth[J]. Cell Res. 2008.18(5):549-57
[36]Slack F.let-7 microRNA reduces tumor growth.Cell Cycle.2009,8(12):1823
[37]Shi G, Perle MA, Mittal K,et al.Let-7 repression leads to HMGA2 overexpression in uterine leiomyosarcoma[J].J Cell Mol Med.2009,(9B):3898-905
[38]Ren Y, Kang CS, Yuan XB, et al.Co-delivery of as-miR-21 and 5-FU by poly(amidoamine) dendrimer attenuates human glioma cell growth in vitro[J].J Biomater Sci Polym Ed. 2010;21(3):303-14
[39]Volinia S, Calin GA, Liu CG, et al.A microRNA expression signature of human solid tumors defines cancer gene targets.Proc Natl Acad Sci U S A.2006,103(7):2257-61
[40]Meng F, Henson R, Wehbe-Janek H, et al. MicroRNA-21 regulates expression of the PTEN tumor suppressor gene in human hepatocellular cancer[J].Gastroenterology.2007133(2): 647-58
[41]Volinia S,Calin GA,Liu C G, et al. A microRNA expression signature of human solid tumors defines cancer gene targets[J],2006,103:2257～2261
[42]Akao Y,Nakagawa Y,Naoe T. Let-7 microRNA functions as a potential growth suppressor in human colon cancer cells[J]. Biol Pharm Bull,2006,29:903～906
[43]SiM L, Zhu S,Wu H, et al. MiR-21-mediated tumor growth[J].2007,26:2799～2803
[44]Kutay H, Bai S, Datta J, et al.Downregulation of miR-122 in the rodent and human hepatocellular carcinomas[J]. J Cell Biochem.2006,99(3):671-8
[45]Pogribny IP, Bagnyukova TV, Tryndyak VP, et al. Gene expression profiling reveals underlying molecular mechanisms of the early stages of tamoxifen-induced rat hepatocarcinogenesis[J].2007,225(1):61-9
[46]Weber F, Teresi RE, Broelsch CE,et al. A limited set of human MicroRNA is deregulated in follicular thyroid carcinoma[J].J Clin Endocrinol Metab.2006,91(9):3584-91
[47][47]Ladeiro Y, Couchy G, Balabaud C,et al.MicroRNA profiling in hepatocellular tumors is associated with clinical features and oncogene/tumor suppressor gene mutations. Hepatology.2008,47(6):1955-63
[48]Murakami Y, Yasuda T, Saigo K,et al.Comprehensive analysis of microRNA expression patterns in hepatocellular carcinoma and non-tumorous tissues[J].Oncogene.2006 ,25(17):2537-45
[49]Gramantieri L, Fornari F, Callegari E,et al.MicroRNA involvement in hepatocellular carcinoma[J].J Cell Mol Med.2008,12(6A):2189-204
[50]Danit Lebanony, Hila Benjamin, Shlomit Gilad Diagnostic Assay Based on hsa-miR-205 Expression Distinguishes Squamous From Nonsquamous Non-Small-Cell Lung Carcinoma[J].Journal of Clinical Oncology, Vol 27, No 12 (April 20),2009:pp.2030-2037
[51]孙小丽,林仲秋,吴维光.上调microRNA-205表达对HeLa细胞生物学特性的影响 中山大学学报(医学科学版),2009/30/02
[52]Geoffrey Childs*, Melissa Fazzari, Low-Level Expression of MicroRNAs let-7d and miR-205 Are Prognostic Markers of Head and Neck Squamous Cell Carcinoma American Journal of Pathology[J].2009; 174:736-745
[53]lorio MV, Casalini P, microRNA-205 regulates HER3 in human breast cancer[J]. Cancer Res.2009 Mar 15;69(6):2195-200
[54]Wu H, Mo YY. Targeting miR-205 in breast cancer Expert Opin Ther Targets[J].2009 Oct 19
[1]Nishida N.Impact of hepatitis virus and aging on DNA methylation in human hepatocarcinogenesis[J].Histol Histopathol.2010,25(5):647-54
[2]Yuan A, Li Z, Li X,et al, The mitogenic effectors of isoproterenol in human hepatocellular carcinoma cells[J].Oncol Rep.2010,23(1):151-7
[3]Bishayee A, Politis T, Darvesh AS.Resveratrol in the chemoprevention and treatment of hepatocellularcarcinoma[J].Cancer Treat Rev.2010,36(1):43-53
[4]Arbuthnot P, Thompson LJ.Harnessing the RNA interference pathway to advance treatment and prevention of hepatocellular carcinoma[J].World J Gastroenterol.2008,14(11):1670-81
[5]Chen Y, Cheng G, Mahato RI.RNAi for treating hepatitis B viral infection[J].Pharm Res. 2008,25(1):72-86
[6]Roessler S, Budhu A, Wang XW.Future of molecular profiling of human hepatocellular carcinoma.Future Oncol[J].2007,3(4):429-39
[7]Lee RC, Feinbaum RL, AmbiosV. The C. elegans gene heterochronic gene lin-4 encodes small RNAs antisense complementarity to lin-14[J].Cell,1993,75(5):843-854
[8]Lagos-Quintana M, Rauhut R, Lendeckel W, et al. Identification of novel genes coding for small expressed RNAs[J]. Science,2001,294:853～858
[9]Lau NC,Lim LP,Weinstein EG,et al.An abundant class of tiny RNAs with probable regulatory roles in Caenorhabditis elegans[J].Science,2001,294(5543):858-862
[10]Rodriguez A,Griffiths-Jones S,Ashurst JL,et al.Identification of mammalian microRNA host genes and transcription units [J].Genome Res,2004,14(10A):1902-1910
[11]Khvorova A,Reynolds A,Jayasena SD.Functional siRNAs and miRNAs exhibit strand bias[J].Cell,2003,115(2):209-216
[12]He L, Hannon GJ. MicroRNAs:small RNAs with a big role in gene regulation[J]. NatRev Genet,2004,5:522～531
[13]Gregory RI, Shiekhattar R. MicroRNA biogenesis and cancer [J]. Cancer Res,2005 ,65(9):3509-12
[14]Hutvagner G,Zamore PD.A microRNA in a multiple-turnover RNAi enzyme complex[J].Science,2002,297(5589):2056-2060
[15]Zeng Y,Yi R,Cullen BR.MicroRNAs and small interfering RNAs can inhibit mRNA expression by similar mechanisms [J].Proc Natl Acad Sci USA,2003,100(17):9779-9784
[16]Lai EC.Micro RNAs are complementary to 3'UTRsequence motifs tha mediate negative post-transcriptional regulation[J].Nat Genet,2002,30(4):363-364
[17]Davidson-Moncada J, Papavasiliou FN, Tam W.MicroRNAs of the immune system: roles in inflammation and cancer[J].Ann N Y Acad Sci.2010,1183:183-94
[18]Zhang H, Li Y, Lai M.The microRNA network and tumor metastasis.Oncogene.2010 29(7):937-48.
[19]Davidson-Moncada J, Papavasiliou FN, Tam W.MicroRNAs of the immune system:roles in inflammation and cancer[J]. Ann N Y Acad Sci.2010,1183:183-94. Review
[20]Shenouda SK, Alahari SK.MicroRNA function in cancer: oncogene or a tumor suppressor[J]? Cancer Metastasis Rev.2009,28(3-4):369-78
[21]Trang P, Weidhaas JB, Slack FJ.MicroRNAs as potential cancer therapeutics[J].Oncogene. 2008, Suppl 2:S52-7
[22]戚鹏,韩金祥,鲁艳芹.疱疹病毒编码的miRNAs的研究进展.中国生物工程杂志,2006,26：69～74
[23]Davalos V, Esteller M.MicroRNAs and cancer epigenetics:a acrorevolution[J].Curr Opin Oncol.2010,22(1):35-45
[24]Taft RJ, Pang KC, Mercer TR, et al, Non-coding RNAs: regulators of disease[J].J Pathol. 2010,220(2):126-39
[25]Baranwal S, Alahari SK.miRNA control of tumor cell invasion and metastasis[J].Int J Cancer.2010,126(6):1283-90
[26]Johnson CD, Esquela-Kerscher A, Stefani G ,et al.The let-7 microRNA represses cell proliferation pathways in human cells[J]. Cancer Res.2007 67(16):7713-22
[27]Chan JA, Krichevsky AM, Kosik KS. MicroRNA-21 is an antiapoptotic factor in human glioblastoma cells[J].Cancer Res.2005;65(14):6029-33
[28]Takagi T, Iio A, Nakagawa Y, Naoe T,et al. Decreased expression of microRNA-143 and-145 in human gastric cancers[J].Oncology,2009,77(1):12-21.
[29]Ren Y, Kang CS, Yuan XB, et al.Co-delivery of as-miR-21 and 5-FU by poly(amidoamine) dendrimer attenuates human glioma cell growth in vitro[J].J Biomater Sci Polym Ed. 2010;21(3):303-14.
[30]Kluiver J, Haralambieva E, de Jong D,et al.Lack of BIC and microRNA miR-155 expression in primary cases of Burkitt lymphoma[J].Genes Chromosomes Cancer.2006,45(2):147-53.
[31]Navon R, Wang H, Steinfeld I,et al. Novel rank-based statistical methods reveal microRNAs with differential expression in multiple cancer types[J].PLoS One.2009,4(11):8003
[32]Calin GA.MicroRNAs and cancer:what we know and what we still have to learn[J].Genome Med.2009,1(8):78
[33]谢纳,生物芯片分析[M].北京：科学出版社,2003
[34]马立人,蒋中华主编,生物芯片[M].北京：化学工业出版社,2000
[35]Pineau P, Volinia S, McJunkin K,et al. miR-221 overexpression contributes to liver tumorigenesis[J].Proc Natl Acad Sci U S A.2010,107(1):264-9
[36]Yamamoto Y, Kosaka N, Tanaka M,Biomarkers.et al. MicroRNA-500 as a potential diagnostic marker for hepatocellular carcinoma[J].2009,14(7):529-38
[37]Wang Y, Lee AT, Ma JZ, et al. Profiling microRNA expression in hepatocellular carcinoma reveals microRNA-224 up-regulation and apoptosis inhibitor-5 as a microRNA-224-specific target[J]. J Biol Chem.2008;283(19):13205-15
[38]Guo Y, Chen Y, Ito H,et al.Identification and characterization of lin-28 homolog B (LIN28B) in human hepatocellular carcinoma[J]. Gene.2006,384:51-61
[39]Huang YS, Dai Y, Yu XF,et al Microarray analysis of microRNA expression in hepatocellular carcinoma and non-tumorous tissues without viral hepatitis[J].J Gastroenterol Hepatol.2008,23(1):87-94
[40]Thorgeirsson SS, Lee JS, Grisham JW.Functional genomics of hepatocellular carcinoma[J]. Hepatology.200643(2 Suppl 1):S145-50
[41]Huang S, He X, Ding J,Upregulation of miR-23a approximately 27a approximately 24 decreases transforming growth factor-beta-induced tumor-suppressive activities in human hepatocellular carcinoma cells[J].Int J Cancer.2008,123(4):972-8
[42]Fornari F, Gramantieri L, Ferracin M, et al.MiR-221 controls CDKN1C/p57 and CDKN1B/p27 expression in human hepatocellular carcinoma[J]. Oncogene.2008,27(43) :5651-61
[43]Datta J, Kutay H, Nasser MW,et al.Methylation mediated silencing ofMicroRNA-1 gene and its role in hepatocellular carcinogenesis[J].Cancer Res.2008,68(13):5049-58
[44]Li W, Xie L, He X,et al Diagnostic and prognostic implications of microRNAs in human hepatocellular carcinoma.Int J Cancer[J].2008,123(7):1616-22
[45]Arbuthnot P, Thompson LJ.Harnessing the RNA interference pathway to advance treatment and prevention of hepatocellular carcinoma[J]. World J Gastroenterol.2008,14(11):1670-81
[46]Sun BS, Dong QZ, Ye QH, et al,Lentiviral-mediated miRNA against osteopontin suppresses tumor growth and metastasis of human hepatocellular carcinoma[J]. Hepatology,48(6): 1834-42
[47]Pineau P, Volinia S, McJunkin K,et al. miR-221 overexpression contributes to liver tumorigenesis[J].Proc Natl Acad Sci U S A.2010,107(1):264-9
[48]Yamamoto Y, Kosaka N, Tanaka M,Biomarkers.et al. MicroRNA-500 as a potential diagnostic marker for hepatocellular carcinoma[J].2009,14(7):529-38
[49]Murakami Y, Yasuda T, Saigo K,et al. Comprehensive analysis of microRNA expression patterns in hepatocellular carcinoma and non-tumorous tissues[J]. Oncogene.2006,25(17): 2537-45
[50]Wang Y, Lee AT, Ma JZ, et al. Profiling microRNA expression in hepatocellular carcinoma reveals microRNA-224 up-regulation and apoptosis inhibitor-5 as a microRNA-224-specific target[J]. J Biol Chem.2008;283(19):13205-15
[51]Kutay H, Bai S, Datta J, et al.Downregulation of miR-122 in the rodent and human hepatocellular carcinomas[J]. J Cell Biochem.2006,99(3):671-8
[52]Meng F, Henson R, Wehbe-Janek H, et al. MicroRNA-21 regulates expression of the PTEN tumor suppressor gene in human hepatocellular cancer[J].Gastroenterology.2007133(2): 647-58
[53]Gramantieri L, Ferracin M, Fornari F,et al.Cyclin G1 is a target of miR-122a, a microRNA frequently down-regulated in human hepatocellular carcinoma[J]. Cancer Res. 2007.67(13):6092-9
[54]Wu X, Wu S, Tong L,et al. miR-122 affects the viability and apoptosis of hepatocellular carcinoma cells[J]. Scand J Gastroenterol.2009;44(11):1332-9
[55]Arbuthnot P, Thompson LJ.Harnessing the RNA interference pathway to advance treatment and prevention of hepatocellular carcinoma[J].World J Gastroenterol.2008,14(11):1670-81
[56]Chen Y, Cheng G, Mahato RI.RNAi for treating hepatitis B viral infection[J].Pharm Res. 2008,25(1):72-86
[57]Ji J, Shi J, Budhu A,et al. MicroRNA expression, survival, and response to interferon in liver cancer[J].N Engl J Med.2009.361(15):1437-47
[58]Budhu A, Jia HL, Forgues M, et al. Identification of metastasis-related microRNAs in hepatocellular carcinoma[J].Hepatology.2008,47(3):897-907
[59]Olson P, Lu J, Zhang H,et al. MicroRNA dynamics in the stages of tumorigenesis correlate with hallmark capabilities of cancer[J]. Genes Dev.2009,23(18):2152-65.
[60]Volinia S,Calin GA,Liu C G, et al. A microRNA expression signature of human solid tumors defines cancer gene targets[J].2006,103:2257～2261
[61]Akao Y,Nakagawa Y,Naoe T. Let-7 microRNA functions as a potential growth suppressor in human colon cancer cells[J]. Biol Pharm Bull,2006,29:903～906
[62]SiM L, Zhu S,Wu H, et al. MiR-21-mediated tumor growth[J].2007,26:2799～2803
[63]Kota J, Chivukula RR, O'Donnell KA,et al. Therapeutic microRNA delivery suppresses tumorigenesis in a murine liver cancer model[J]. Cell.2009.137(6):1005-17.