[1]葛乔枫,张龙,孙晓娟,等.敲低核转运蛋白对人骨肉瘤细胞增殖和血管生成的体内影响[J].第三军医大学学报,2019,41(14):1308-1313.
 GE Qiaofeng,ZHANG Long,SUN Xiaojuan,et al.Karyopherina 2 knockdown inhibits osteosarcoma cell proliferation and angiogenesis in mice by suppressing Sox2 expression[J].J Third Mil Med Univ,2019,41(14):1308-1313.
点击复制

敲低核转运蛋白对人骨肉瘤细胞增殖和血管生成的体内影响(/HTML )
分享到:

《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
41卷
期数:
2019年第14期
页码:
1308-1313
栏目:
基础医学
出版日期:
2019-07-30

文章信息/Info

Title:
Karyopherina 2 knockdown inhibits osteosarcoma cell proliferation and angiogenesis in mice by suppressing Sox2 expression
作者:
葛乔枫张龙孙晓娟吕智
山西医科大学第二临床医学院骨科
Author(s):
Department of Orthopedics, Second Hospital of Shanxi Medical University, Taiyuan, Shanxi Province, 030000, China
关键词:
核转运蛋白骨肉瘤细胞细胞增殖肿瘤血管生成Sox2裸鼠
Keywords:
karyopherina 2  osteosarcoma cell cell proliferation tumor angiogenesis Sox2 nude mice
分类号:
R394.3;R730.23;R738.1
文献标志码:
A
摘要:

目的 探讨核转运蛋白2(karyopherina 2,KPNA2)对人骨肉瘤(osteosarcoma,OS)细胞增殖和血管生成的影响,并从分子层面分析其可能的作用机制。方法 构建Sh-KPNA2慢病毒载体(Sh-KPNA2组)及空载体Sh-Control(Sh-Control组)转染骨肉瘤细胞Saos-2,观察敲低KPNA2后对Saos-2细胞增殖的影响;利用转染Sh-KPNA2和空载体Sh-Control的Saos-2细胞分别构建OS动物模型,小动物活体荧光分子断层成像系统(fluorescence molecular tomography,FMT)验证KPNA2对瘤体血管生成的影响;qRT-PCR和Western blot验证瘤体内KPNA2和睾丸决定因子Y染色体性别决定区基因(SRY related HMG box-2,Sox2)的mRNA和蛋白表达水平。结果 Sh-KPNA2组细胞增殖能力较Sh-Control组明显降低(P<0.05);Sh-KPNA2组荷瘤小鼠瘤体积显著小于Sh-Control组(P<0.05);免疫组化和FMT扫描结果显示,Sh-Control组的血管生成能力显著高于Sh-KPNA2组(P<0.05);瘤体qRT-PCR和Western blot检测结果均显示Sox2在Sh-Control组中表达显著高于Sh-KPNA2组(P<0.05)。结论 敲低KPNA2可抑制OS增殖和血管生成,其机制可能与KPNA2影响Sox2的表达有关。

Abstract:

Objective To explore the effect of karyopherina 2 (KPNA2) knockdown on osteosarcoma cell proliferation and angiogenesis and the possible molecular mechanism. Methods We transfected Saos-2 cells with a Sh-KPNA2 lentiviral vector or a control vector (Sh-Control) to observe the effect of KPNA2 knockdown on the cell proliferation. We further observed tumor angiogenesis in nude mice bearing xenografts derived from Saos-2 cells transfected with Sh-KPNA2 or Sh-Control vector using a fluorescence molecular tomography (FMT) system. qRT-PCR and Western blotting were performed to detect the mRNA and protein expression of KPNA2 and SRY-related HMG box-2 (Sox2) in the xenografts. ResultsSaos-2 cells transfected with Sh-KPNA2 vector showed significantly lowered proliferation capacity as compared with the cells transfected with Sh-Control vector (P<0.05). In the tumor-bearing nude mice, the tumor volume and weight were both significantly lower in Sh-KPNA2 group than in Sh-Control group (P<0.05). The results of FMT scan and immunohistochemistry both showed weakened angiogenesis in the xenografts in Sh-KPNA2 group in comparison with Sh-Control group (P<0.05). Sox2 expression at both the mRNA and protein levels in the xenografts was significantly lower in Sh-KPNA2 group than in Sh-Control group (P<0.05). Conclusion KPNA2 knockdown inhibits the proliferation and angiogenesis of osteosarcoma cells possibly as a result of lowered expression of Sox2.

参考文献/References:

[1]LUETKE A, MEYERS P A, LEWIS I, et al. Osteosarcoma treatment——where do we stand? A state of the art review[J]. Cancer Treat Rev, 2014, 40(4): 523-532. DOI: 10.1016/j.ctrv.2013.11.006.
[2]FERRARI S, MEAZZA C, PALMERINI E, et al.Nonmetastatic osteosarcoma of the extremity. Neoadjuvant chemotherapy with methotrexate, cisplatin, doxorubicin and ifosfamide. An Italian Sarcoma Group study (ISG/OS-Oss)[J]. Tumori, 2014, 100(6): 612-619. DOI:10.1700/1778.19262.
[3]BULUT G, HONG S H, CHEN K, et al. Small molecule inhibitors of ezrin inhibit the invasive phenotype of osteosarcoma cells[J]. Oncogene, 2012, 31(3): 269-281. DOI: 10.1038/onc.2011.245. 
[4]DURFEE R A, MOHAMMED M, LUU H H. Review of osteosarcoma and current management[J]. Rheumatol Ther, 2016, 3(2): 221-243. DOI: 10.1007/s40744-016-0046-y. 
[5]SCHWARTZ B, BENADJAOUD M A, CLRO E, et al. Risk of second bone sarcoma following childhood cancer: role of radiation therapy treatment[J]. Radiat Environ Biophys, 2014, 53(2): 381-390. DOI: 10.1007/s00411-013-0510-9. 
[6]VERHAAK R G, GOUDSWAARD C S, VAN PUTTEN W, et al. Mutations in nucleophosmin (NPM1) in acute myeloid leukemia (AML): association with other gene abnormalities and previously established gene expression signatures and their favorable prognostic significance[J]. Blood, 2005, 106(12): 3747-3754. DOI: 10.1182/blood-2005-05-2168. 
[7]HUANG L, WANG H Y, LI J D, et al. KPNA2 promotes cell proliferation and tumorigenicity in epithelial ovarian carcinoma through upregulation of c-Myc and downregulation of FOXO3a [J]. Cell Death Dis, 2013, 4(8): e745. DOI:10.1038/cddis.2013.256.
[8]HU Z Y, YUAN S X, YANG Y, et al. Pleomorphic adenoma gene 1 mediates the role of karyopherin alpha 2 and has prognostic significance in hepatocellular carcinoma[J]. J Exp Clin Cancer Res, 2014, 33: 61. DOI: 10.1186/s13046-014-0061-1. 
[9]LIN J X, ZHANG L, HUANG H, et al. MiR-26b/KPNA2 axis inhibits epithelial ovarian carcinoma proliferation and metastasis throughdownregulating OCT4[J]. Oncotarget, 2015, 6(27): 23793-23806. DOI: 10.18632/oncotarget.4363. 
[10]LI X L, JIA L L, SHI M M, et al. Downregulation of KPNA2 in non-small-cell lung cancer is associated with Oct4 expression[J]. J Transl Med, 2013, 11(1): 232. DOI: 10.1186/1479-5876-11-232. 
[11]BLATTNER M, LEE D J, O’REILLY C, et al. SPOP mutations in prostate cancer across demographically diverse patient cohorts[J].Neoplasia, 2014, 16(1): 14-W10. DOI: 10.1593/neo.131704. 
[12]WANG C I, CHEN YY, WANG C L, et al. mTOR regulates proteasomal degradation and Dp1/E2F1-mediated transcription of KPNA2 in lung cancer cells [J]. Oncotarget, 2016, 7(18): 25432-25442. DOI:10.18632/oncotarget.8170.
[13]FAES S, SANTORO T, DEMARTINES N, et al. Evolving significance and future relevance of anti-angiogenic activity of mTOR inhibitors in cancer therapy[J]. Cancers (Basel), 2017, 9(11): E152. DOI: 10.3390/cancers9110152. 
[14]AUFFINGER B, TOBIAS A L, HAN Y, et al. Conversion of differentiated cancer cells into cancer stem-like cells in aglioblastoma model after primary chemotherapy[J]. Cell Death Differ, 2014, 21(7): 1119-1131. DOI: 10.1038/cdd.2014.31. 
[15]MARTINS-NEVES S R, CORVER W E, PAIVA-OLIVEIRA D I, et al. Osteosarcoma stem cells have active wnt/β-catenin and overexpress SOX2 and KLF4[J]. J Cell Physiol, 2016, 231(4): 876-886. DOI: 10.1002/jcp.25179. 
[16]MAURIZI G, VERMA N, GADI A, et al. Sox2 is required for tumor development and cancer cell proliferation in osteosarcoma[J]. Oncogene, 2018, 37(33): 4626-4632. DOI: 10.1038/s41388-018-0292-2.
 

更新日期/Last Update: 2019-07-22