miR-455-5p在结直肠癌中的表达及生物学特性
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摘要
目的探讨miR-455-5p在结直肠癌(CRC)中的表达及生物学特性。方法收集40例CRC患者经手术切除的结肠和直肠癌组织标本及癌旁10cm的正常黏膜组织标本。采用qRT-PCR法检测CRC组织及癌旁正常黏膜组织中miR-455-5p的表达水平;采用脂质体转染试剂转染CRC细胞,Brdu增殖实验检测转染后CRC细胞增殖能力,流式细胞仪检测转染后CRC细胞凋亡率。应用生物信息学方法预测并验证miR-455-5p的靶基因。结果 CRC组织miR-455-5p的相对表达量低于癌旁正常黏膜组织(P<0.01)。经转染后CRC细胞中,上调miR-455-5p可抑制CRC细胞增殖和迁移,促进细胞凋亡。数据库预测PIK3R1可能是miR-455-5p的靶基因,上调miR-455-5p可抑制PIK3R1mRNA及蛋白表达。结论 miR-455-5p在CRC组织中呈低表达,其生物学特性是miR-455-5p具有抑制CRC细胞生长的作用,其机制可能与调控PIK3R1的表达有关。
Objective To investigate the expression of miR-455-5p in colorectal carcinoma and its relation to biological features of cancer. Methods The surgical specimens of cancer tissue and pericancerous mucosa tissue were collected from 40 cases of colorectal carcinoma. The expression of miR-455-5p was determined by qRT-PCR in the cancer tissue and pericancerous tissue. The colorectal cancer SW-480, HCT-116, HT-29 cells were transfected with miR-455-5p-mimic or inhibitor, respectively. The cell growth and proliferation capacity were assayed by Brdu method; the cell apoptosis was tested by flow cytometry; the target genes of miR-455-5p were predicted with bioinformatic analysis. Results The expression of miR-455-5p was down-regulation in colorectal cancer tissue compared with pericancerous tissue. In the colorectal cancer cells transfected with miR-455-5p-mimic, the expression of miR-455-5p was up-regulated, and the cell proliferation and migration were inhibited, and apoptosis was promoted. Bioinformatic analysis showed that PIK3R1 might be the target gene of miR-455-5p. The up-regulation of miR-455-5 p inhibited the expression of PIK3R1 mRNA and protein. Conclusion The expression of miR-455-5p is down-regulated in colorectal cancer, indicating that miR-455-5p may inhibit the growth of CRC cells, which may be related to the regulation of PIK3R1 expression.
Objective To investigate the expression of miR-455-5p in colorectal carcinoma and its relation to biological features of cancer. Methods The surgical specimens of cancer tissue and pericancerous mucosa tissue were collected from 40 cases of colorectal carcinoma. The expression of miR-455-5p was determined by qRT-PCR in the cancer tissue and pericancerous tissue. The colorectal cancer SW-480, HCT-116, HT-29 cells were transfected with miR-455-5p-mimic or inhibitor, respectively. The cell growth and proliferation capacity were assayed by Brdu method; the cell apoptosis was tested by flow cytometry; the target genes of miR-455-5p were predicted with bioinformatic analysis. Results The expression of miR-455-5p was down-regulation in colorectal cancer tissue compared with pericancerous tissue. In the colorectal cancer cells transfected with miR-455-5p-mimic, the expression of miR-455-5p was up-regulated, and the cell proliferation and migration were inhibited, and apoptosis was promoted. Bioinformatic analysis showed that PIK3R1 might be the target gene of miR-455-5p. The up-regulation of miR-455-5 p inhibited the expression of PIK3R1 mRNA and protein. Conclusion The expression of miR-455-5p is down-regulated in colorectal cancer, indicating that miR-455-5p may inhibit the growth of CRC cells, which may be related to the regulation of PIK3R1 expression.
引文
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[3]Brenner H,Stock C,Hoffmeister M.Colorectal cancer screening:the time to act is now[J].BMC Med,2015,13:262-266.doi:10.1186/s12916-015-0498-x.
[4]Lee RC,Feinbaum RL,Ambros V.The C.elegans heterochronic gene lin-4 encodes small rnas with antisense complementarity to Lin-14[J].Cell,1993,75:843-854.
[5]Zoni E,van der Pluijm G,Gray PC,et al.Epithelial Plasticity in Cancer:Unmasking a MicroRNA Network for TGF-β-,Notch-,and Wnt-Mediated EMT[J].J Oncol,2015,198967:1-13.doi:10.1155/2015/198967.
[6]Kohlhapp FJ,Mitra AK,Lengyel E,et al.MicroRNAs as mediators and communicators between cancer cells and the tumor microenvironment[J].Oncogene,2015,34(48):5857-5868.doi:10.1038/onc.2015.89.
[7]Yi R,Li Y,Wang FL,et al.MicroRNAs as diagnostic and prognostic biomarkers in colorectal cancer[J].World J Gastrointest Oncol,2016,8(4):330-340.doi:10.4251/wjgo.v8.i4.330.
[8]Hudson J,Duncavage E,Tamburrino,et al.Overexpression of miR-10a and miR-375 and downregulation of YAP1 in medullary thyroid carcinoma[J].Exp Mol Pathol,2013,95:62-67.doi:10.1016/j.yexmp.2013.05.001.
[9]Shoshan E,Mobley AK,Braeuer RR,et al.Reduced adenosineto-inosine miR-455-5p editing promotes melanoma growth and metastasis[J].Nat Cell Biol,2015,17:311-321.doi:10.1038/ncb3110.
[10]Liu J,Zhang J,Li Y,et al.MiR-455-5p acts as a novel tumor suppressor in gastric cancer by down-regulating RAB18[J].Gene,2016,592(2):308-315.doi:10.1016/j.gene.2016.07.034.
[11]Peeters M,Price T.Biologic therapies in the metastatic colorectal cancer treatment continuum-applying current evidence to clinical practice[J].Cancer Treatment Reviews,2012,38(5):397-406.doi:10.1016/j.ctrv.2011.08.002.
[12]Morillas JD,Castells A,Oriol I,et al.The Alliance for the Prevention of Colorectal Cancer in Spain.A civil commitment to society[J].GastroenterolHepatolog,2012,35(3):109-128.doi:10.1016/j.gastrohep.2012.01.002.
[13]He K,Jin K,Wang H,et al.Anti-angiogenic therapy for colorectal cancer:on the way to getting better[J].Hepatogastroenterology,2012,59(116):1113-1117.doi:10.5754/hge10722.
[14]Ng F,Ganeshan B,Kozarski R,et al.Assessment of Primary Colorectal Cancer Heterogeneity by Using Whole-Tumor Texture Analysis:Contrast-enhanced CT Texture as a Biomarker of5-year Survival[J].Radiology,2012,266(1):177-184.doi:10.1148/radiol.12120254.
[15]Troiani T,Martinelli E,Orditura M,et al.Beyond bevacizumab:new anti-VEGF strategies in colorectal cancer[J].Expert Opinion on Investigational Drugs,2012,21(7):949-959.doi:10.1517/13543784.2012.689287.
[16]Toiyama Y,Tanaka K,Inoue Y,et al.Circulating cell-free microRNAs as biomarkers for colorectal cancer[J].Surgery Today,2016,46(1):13-24.doi:10.1007/s00595-015-1138-y.
[17]Slaby O,Svoboda M,Fabian P,et al.Altered expression of miR-21,miR-31,miR-143 and miR-145 is related to clinicopathologic features of colorectal cancer[J].Oncology,2007,72(5-6):397-402.doi:10.1159/000113489.
[18]Yang Q,Hou C,Huang D,et al.miR-455-5p functions as a potential oncogene by targeting galectin-9 in colon cancer[J].Oncology Letters,2017,13(3):1958-1964.doi:10.3892/ol.2017.5608.
[19]Cizkova M,Vacher S,Meseure D,et al.PIK3R1 underexpression is an independent prognostic marker in breast cancer[J].BMCCancer,2013,13(1):1-15.doi:10.1186/1471-2407-13-545.
[20]Lee SR,Kim SJ,Park Y,et al.Bortezomib and melphalan as a conditioning regimen for autologous stem cell transplantation in multiple myeloma[J].Korean Journal of Hematology,2010,45(3):183-187.doi:10.5045/kjh.2010.45.3.183.
[21]Qiao M,Sheng S,Pardee AB.Metastasis and AKT activation[J].802JournalofCellularPhysiology,2008,7(19):2991-2996.doi:10.4161/cc.7.19.6784.
[22]Kaur J.PI3-kinase/Wnt association mediates COX-2/PGE(2)pathway to inhibit apoptosis in early stages of colon carcinogenesis:chemoprevention by diclofenac[J].Tumour Biology the Journal of the International Society for Oncodevelopmental Biology&Medicine,2010,31(6):623-631.doi:10.1007/s13277-010-0078-9.