miR-548d抑制骨肉瘤细胞的增殖和迁移
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摘要
目的:探讨miR-548d对骨肉瘤细胞增殖和迁移的影响。方法:通过Real time PCR及Western blot检测miR-548d和KRAS在30例骨肉瘤组织以及293、MG63和U2OS细胞中的表达情况。对30例骨肉瘤组织中miR-548d和KRAS含量的相关性进行分析,随后通过报告基因实验印证miR-548d对KRAS的靶向作用。MG63细胞中分别过表达或沉默miR-548d后,通过Western blot实验分析KRAS的变化情况,MTT实验观察miR-548d对细胞增殖的影响,Transwell实验观察miR-548d对其迁移能力的影响。结果:骨肉瘤组织及细胞中miR-548d表达较低,KRAS表达较高。在骨肉瘤组织中miR-548d与KRAS的表达呈负相关。报告基因实验证明miR-548d可以直接打靶KRAS。Western blot指出miR-548d可以抑制KRAS的表达。最后MTT和Transwell实验指出miR-548d可以抑制MG63细胞的增殖与迁移。结论:miR-548d可以通过打靶KRAS抑制骨肉瘤细胞的增殖和迁移。
Objective:To investigate the effect of miR-548d on the proliferation and migration of osteosarcoma.Methods:The expression of miR-548d and KRAS in 30 osteosarcoma tissues and 293,MG63 and U2OS cells were detected by Real time PCR and Western blot.The correlation of the content of miR-548d and KRAS in 30 cases of osteosarcoma was analyzed,and then the target effect of miR-548d on KRAS was confirmed by the reported gene experiment.After overexpressing or silencing miR-548d in MG63 cells,the change of KRAS was analyzed by Western blot.The effect of miR-548d on cell proliferation was observed by MTT,and the effect of miR-548d on migration ability was observed by Transwell.Results:The expression of miR-548d in the tissues and cells of osteosarcoma was lower,and the expression of KRAS was higher.There was a negative correlation between the expression of miR-548d and KRAS in osteosarcoma.The report gene experiment showed that miR-548d could target KRAS directly.Western blot points out that miR-548d could inhibit the expression of KRAS.Finally,MTT and transwell showed that miR-548d could inhibit the proliferation and migration of MG63 cells.Conclusion:miR-548d can inhibit the proliferation and migration of osteosarcoma cells by targeting KRAS.
Objective:To investigate the effect of miR-548d on the proliferation and migration of osteosarcoma.Methods:The expression of miR-548d and KRAS in 30 osteosarcoma tissues and 293,MG63 and U2OS cells were detected by Real time PCR and Western blot.The correlation of the content of miR-548d and KRAS in 30 cases of osteosarcoma was analyzed,and then the target effect of miR-548d on KRAS was confirmed by the reported gene experiment.After overexpressing or silencing miR-548d in MG63 cells,the change of KRAS was analyzed by Western blot.The effect of miR-548d on cell proliferation was observed by MTT,and the effect of miR-548d on migration ability was observed by Transwell.Results:The expression of miR-548d in the tissues and cells of osteosarcoma was lower,and the expression of KRAS was higher.There was a negative correlation between the expression of miR-548d and KRAS in osteosarcoma.The report gene experiment showed that miR-548d could target KRAS directly.Western blot points out that miR-548d could inhibit the expression of KRAS.Finally,MTT and transwell showed that miR-548d could inhibit the proliferation and migration of MG63 cells.Conclusion:miR-548d can inhibit the proliferation and migration of osteosarcoma cells by targeting KRAS.
引文
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[13] GM Roberto,EE Engel,CA Scrideli,et al.Downregulation of miR-10B* is correlated with altered expression of mitotic kinases in osteosarcoma[J].Pathology,Research and Practice,2018,214(2):213-216.
[14] CJ Rushing,DE Rogers,SM Spinner,et al.A case report of heel pain mimicking plantar fasciitis and osteosarcoma:A unique presentation of a Nora's lesion[J].The Journal of Foot and Ankle Surgery,2017,56:670-673.
[15] I Saadon,B Amit,A Zolquarnian,et al.Primary osteosarcoma of the distal fibula treated with distal fibulectomy with a five-year follow-up:A case report[J].Malaysian Orthopaedic Journal,2017,11:64-67.
[16] Y Sahin,Z Altan,K Arman,et al.Inhibition of miR-664a interferes with the migration of osteosarcoma cells via modulation of MEG3[J].Biochemical and Biophysical Research Communications,2017,490:1100-1105.
[2] VL Rizzo,CB Levine,JJ Wakshlag.The effects of sulforaphane on canine osteosarcoma growth and invasion[J].Veterinary and Comparative Oncology,2017,15:718-730.
[3] GB Andersen,A Knudsen,H Hager,et al.miRNA profiling identifies deregulated miRNAs associated with osteosarcoma development and time to metastasis in two large cohorts[J].Molecular Oncology,2018,12:114-131.
[4] T Luo,X Yi,W Si.Identification of miRNA and genes involving in osteosarcoma by comprehensive analysis of microRNA and copy number variation data[J].Oncology Letters,2017,14:5427-5433.
[5] H Ke,L Zhao,X Feng,et al.NEAT1 is required for survival of breast cancer cells through FUS and miR-548[J].Gene Regulation and Systems Biology,2016,10:11-17.
[6] H Zhang,QY He,GC Wang,et al.miR-422a inhibits osteosarcoma proliferation by targeting BCL2L2 and KRAS[J].Bioscience Reports,2018,38(2):BSR20170339.
[7] S Zhang,C Hou,G Li,et al.A single nucleotide polymorphism in the 3'-untranslated region of the KRAS gene disrupts the interaction with let-7a and enhances the metastatic potential of osteosarcoma cells[J].International Journal of Molecular Medicine,2016,38:919-926.
[8] X Zhang,Q Guo,J Chen,et al.Quercetin enhances cisplatin sensitivity of human osteosarcoma cells by modulating microRNA-217-KRAS axis[J].Molecules and Cells,2015,38:638-642.
[9] S Zhu,C He,S Deng,et al.MiR-548an,transcriptionally downregulated by HIF1alpha/HDAC1,suppresses tumorigenesis of pancreatic cancer by targeting Vimentin expression[J].Molecular Cancer Therapeutics,2016,15:2209-2219.
[10] S Sanghavi,A Wahegaonkar,Y Panchwagh,et al.Parosteal osteosarcoma of the distal radius mimicking an osteochondroma-A diagnostic misadventure[J].The Journal of Hand Surgery,2017,42:1038.e1-1038.e10.
[11] A Sasikumar,A Joy,MRA Pillai,et al.68Ga-PSMA PET/CT in osteosarcoma in fibrous dysplasia[J].Clinical Nuclear Medicine,2017,42:446-447.
[12] SA Savage,L Mirabello.Bone cancer:Is the osteosarcoma genome targetable[J]?Nature Reviews Endocrinology,2017,13:506-508.
[13] GM Roberto,EE Engel,CA Scrideli,et al.Downregulation of miR-10B* is correlated with altered expression of mitotic kinases in osteosarcoma[J].Pathology,Research and Practice,2018,214(2):213-216.
[14] CJ Rushing,DE Rogers,SM Spinner,et al.A case report of heel pain mimicking plantar fasciitis and osteosarcoma:A unique presentation of a Nora's lesion[J].The Journal of Foot and Ankle Surgery,2017,56:670-673.
[15] I Saadon,B Amit,A Zolquarnian,et al.Primary osteosarcoma of the distal fibula treated with distal fibulectomy with a five-year follow-up:A case report[J].Malaysian Orthopaedic Journal,2017,11:64-67.
[16] Y Sahin,Z Altan,K Arman,et al.Inhibition of miR-664a interferes with the migration of osteosarcoma cells via modulation of MEG3[J].Biochemical and Biophysical Research Communications,2017,490:1100-1105.