miR-140-5p在骨肉瘤中的表达及其对骨肉瘤细胞侵袭转移的影响
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摘要
目的研究miR-140-5p在骨肉瘤中的表达及其对骨肉瘤细胞MG-63及U2OS侵袭转移的影响。方法 qRT-PCR及免疫原位杂交法检测miR-140-5p在骨肉瘤组织中的表达情况; qRT-PCR检测miR-140-5p在骨肉瘤细胞M G-63、U2OS及正常成骨细胞系hFOB 1. 19中的表达;在MG-63、U2OS细胞中转染miR-140-5p mimic以上调miR-140-5p的表达,qRT-PCR确认转染成功; Transwell实验检测不同miR-140-5p表达水平对骨肉瘤细胞MG-63、U2OS侵袭转移的影响。结果与癌旁组织比较,miR-140-5p在骨肉瘤组织中表达降低;与正常成骨细胞系h FOB 1. 19比较,miR-140-5p在骨肉瘤细胞MG-63、U2OS中表达降低;在MG-63、U2OS中转染miR-140-5p mimic后,miR-140-5p表达升高,M G-63、U2OS细胞侵袭转移能力下降。结论 miR-140-5p在骨肉瘤组织及细胞系MG-63、U2OS中表达下降,上调其表达水平可抑制MG-63、U2OS细胞侵袭转移。
Objective To explore the expression of miR-140-5p in osteosarcoma and its influence on the invasion and migration of MG-63 and U2OS. Methods The expression of miR-140-5p in osteosarcoma tissue specimens was determined by using qRT-PCR assay and an in situ hybridization assay. The expression of miR-140-5p in MG-63,U2OS and h FOB1. 19 was measured by qRT-PCR assay. miR-140-5p mimics were transfected into MG-63 and U2OS cells and were confirmed by a qRT-PCR assay to gain a miR-140-5p over-expressed cell model. Transwell assays were executed to evaluate the influence of expression of miR-140-5p on the migration and invasion of MG-63 and U2OS cells. Results The expression of miR-140-5p was down-regulated in osteosarcoma tissue specimens compared with that in para-tumor tissue specimens,and it was also down-regulated in osteosarcoma cell lines MG-63 and U2OS compared with that of h FOB1. 19. The expression of miR-140-5p was up-regulated in MG-63 and U2OS cells which were transfected with miR-140-5p mimics,then the ability of invasion and migration of MG-63 and U2OS cells was decreased. Conclusion The expression of miR-140-5p is down-regulated in osteosarcoma and cell lines MG-63 and U2OS. Up-regulating miR-140-5p can suppress the invasion and migration of MG-63 and U2OS cells.
Objective To explore the expression of miR-140-5p in osteosarcoma and its influence on the invasion and migration of MG-63 and U2OS. Methods The expression of miR-140-5p in osteosarcoma tissue specimens was determined by using qRT-PCR assay and an in situ hybridization assay. The expression of miR-140-5p in MG-63,U2OS and h FOB1. 19 was measured by qRT-PCR assay. miR-140-5p mimics were transfected into MG-63 and U2OS cells and were confirmed by a qRT-PCR assay to gain a miR-140-5p over-expressed cell model. Transwell assays were executed to evaluate the influence of expression of miR-140-5p on the migration and invasion of MG-63 and U2OS cells. Results The expression of miR-140-5p was down-regulated in osteosarcoma tissue specimens compared with that in para-tumor tissue specimens,and it was also down-regulated in osteosarcoma cell lines MG-63 and U2OS compared with that of h FOB1. 19. The expression of miR-140-5p was up-regulated in MG-63 and U2OS cells which were transfected with miR-140-5p mimics,then the ability of invasion and migration of MG-63 and U2OS cells was decreased. Conclusion The expression of miR-140-5p is down-regulated in osteosarcoma and cell lines MG-63 and U2OS. Up-regulating miR-140-5p can suppress the invasion and migration of MG-63 and U2OS cells.
引文
[1]Berner K,Johannesen TB,Berner A,et al.Time-trends on incidence and survival in a nationw ide and unselected cohort of patients w ith skeletal osteosarcoma[J].Acta Oncol,2015,54(1):25-33.
[2]Lee L,Fei L,Pope J,et al.Early lymphocyte recovery and outcome in osteosarcoma[J].J Pediatr Hematol Oncol,2017,39(3):179-183.
[3]Ottaviani G,Jaffe N.The epidemiology of osteosarcoma[J].Cancer Treatment Res,2009,152:3-13.
[4]Serlo J,Tarkkanen M,Sampo M,et al.Incidence,treatment and survival of paediatric patients w ith bone sarcomas in Finland from 1991 to 2005[J].Acta Paediatr,2015,104(7):738-745.
[5]Friebele JC,Peck J,Pan X,et al.Osteosarcoma:a Meta-analysis and review of the literature[J].Am J Orthop(Belle M ead NJ),2015,44(12):547-553.
[6]He X,Gao Z,Xu H,et al.A meta-analysis of randomized control trials of surgical methods w ith osteosarcoma outcomes[J].J Orthop Surg Res,2017,12(1):5.
[7]Eckstein F.Small non-coding RNAs as magic bullets[J].Trends Biochem Sci,2005,30(8):445-452.
[8]Wu W,Sun M,Zou GM,et al.MicroRNA and cancer:current status and prospective[J].Int J Cancer,2007,120(5):953-960.
[9]Fang Z,Yin S,Sun R,et al.miR-140-5p suppresses the proliferation,migration and invasion of gastric cancer by regulating YES1[J].M ol Cancer,2017,16(1):139.
[10]Flamini V,Jiang WG,Cui Y.Therapeutic role of MiR-140-5p for the treatment of non-small cell lung cancer[J].Anticancer Res,2017,37(8):4319-4327.
[11]Ingelmo-Torres M,Lozano JJ,Izquierdo L,et al.Urinary cell microRNA-based prognostic classifier for non-muscle invasive bladder cancer[J].Oncotarget,2017,8(11):18238-18247.
[12]Marchese FP,Huarte M.Long non-coding RNAs and chromatin modifiers:their place in the epigenetic code[J].Epigenetics,2014,9(1):21-26.
[13]Hausser J,Zavolan M.Identification and consequences of miR-NA-target interactions--beyond repression of gene expression[J].Nat Rev Genet,2014,15(9):599-612.
[14]Cha W,Fan R,Miao Y,et al.MicroRNAs as novel endogenous targets for regulation and therapeutic treatments[J].M edchemcomm,2018,9(3):396-408.
[15]Bhardwaj A,Singh S,Singh AP.MicroRNA-based cancer therapeutics:big hope from small RNAs[J].M ol Cell Pharmacol,2010,2(5):213-219.
[16]Zhang W,Zou C,Pan L,et al.MicroRNA-140-5p inhibits the progression of colorectal cancer by targeting VEGFA[J].Cell Physiol Biochem,2015,37(3):1123-1133.
[17]Lan H,Chen W,He G,et al.MiR-140-5p inhibits ovarian cancer grow th partially by repression of PDGFRA[J].Biomed Pharmacother,2015,75:117-122.
[18]Jing P,Sa N,Liu X,et al.MicroR-140-5p suppresses tumor cell migration and invasion by targeting ADAM10-mediated Notch1signaling pathw ay in hypopharyngeal squamous cell carcinoma[J].Exp Mol Pathol,2016,100(1):132-138.
[19]Yang H,Fang F,Chang R,et al.MicroRNA-140-5p suppresses tumor grow th and metastasis by targeting transforming grow th factorβreceptor 1 and fibroblast grow th factor 9 in hepatocellular carcinoma[J].Hepatology,2013,58(1):205-217.
[2]Lee L,Fei L,Pope J,et al.Early lymphocyte recovery and outcome in osteosarcoma[J].J Pediatr Hematol Oncol,2017,39(3):179-183.
[3]Ottaviani G,Jaffe N.The epidemiology of osteosarcoma[J].Cancer Treatment Res,2009,152:3-13.
[4]Serlo J,Tarkkanen M,Sampo M,et al.Incidence,treatment and survival of paediatric patients w ith bone sarcomas in Finland from 1991 to 2005[J].Acta Paediatr,2015,104(7):738-745.
[5]Friebele JC,Peck J,Pan X,et al.Osteosarcoma:a Meta-analysis and review of the literature[J].Am J Orthop(Belle M ead NJ),2015,44(12):547-553.
[6]He X,Gao Z,Xu H,et al.A meta-analysis of randomized control trials of surgical methods w ith osteosarcoma outcomes[J].J Orthop Surg Res,2017,12(1):5.
[7]Eckstein F.Small non-coding RNAs as magic bullets[J].Trends Biochem Sci,2005,30(8):445-452.
[8]Wu W,Sun M,Zou GM,et al.MicroRNA and cancer:current status and prospective[J].Int J Cancer,2007,120(5):953-960.
[9]Fang Z,Yin S,Sun R,et al.miR-140-5p suppresses the proliferation,migration and invasion of gastric cancer by regulating YES1[J].M ol Cancer,2017,16(1):139.
[10]Flamini V,Jiang WG,Cui Y.Therapeutic role of MiR-140-5p for the treatment of non-small cell lung cancer[J].Anticancer Res,2017,37(8):4319-4327.
[11]Ingelmo-Torres M,Lozano JJ,Izquierdo L,et al.Urinary cell microRNA-based prognostic classifier for non-muscle invasive bladder cancer[J].Oncotarget,2017,8(11):18238-18247.
[12]Marchese FP,Huarte M.Long non-coding RNAs and chromatin modifiers:their place in the epigenetic code[J].Epigenetics,2014,9(1):21-26.
[13]Hausser J,Zavolan M.Identification and consequences of miR-NA-target interactions--beyond repression of gene expression[J].Nat Rev Genet,2014,15(9):599-612.
[14]Cha W,Fan R,Miao Y,et al.MicroRNAs as novel endogenous targets for regulation and therapeutic treatments[J].M edchemcomm,2018,9(3):396-408.
[15]Bhardwaj A,Singh S,Singh AP.MicroRNA-based cancer therapeutics:big hope from small RNAs[J].M ol Cell Pharmacol,2010,2(5):213-219.
[16]Zhang W,Zou C,Pan L,et al.MicroRNA-140-5p inhibits the progression of colorectal cancer by targeting VEGFA[J].Cell Physiol Biochem,2015,37(3):1123-1133.
[17]Lan H,Chen W,He G,et al.MiR-140-5p inhibits ovarian cancer grow th partially by repression of PDGFRA[J].Biomed Pharmacother,2015,75:117-122.
[18]Jing P,Sa N,Liu X,et al.MicroR-140-5p suppresses tumor cell migration and invasion by targeting ADAM10-mediated Notch1signaling pathw ay in hypopharyngeal squamous cell carcinoma[J].Exp Mol Pathol,2016,100(1):132-138.
[19]Yang H,Fang F,Chang R,et al.MicroRNA-140-5p suppresses tumor grow th and metastasis by targeting transforming grow th factorβreceptor 1 and fibroblast grow th factor 9 in hepatocellular carcinoma[J].Hepatology,2013,58(1):205-217.