1.400010 重庆,重庆医科大学附属第二医院骨科
2.400037 重庆,陆军军医大学(第三军医大学)第二附属医院病理科
Received:05 December 2019,
Revised:2020-03-09,
Published:30 June 2020
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Lifeng GUO, Yangfan LYU, Ya CAO, et al. Ran-binding protein 9 overexpression induces apoptosis in osteosarcoma cells by activating endogenous mitochondrial pathway[J]. Journal of Third Military Medical University, 2020, 42(12): 1174-1181.
Lifeng GUO, Yangfan LYU, Ya CAO, et al. Ran-binding protein 9 overexpression induces apoptosis in osteosarcoma cells by activating endogenous mitochondrial pathway[J]. Journal of Third Military Medical University, 2020, 42(12): 1174-1181. DOI: 10.16016/j.1000-5404.201912045.
目的
2
探讨过表达RanBP9对骨肉瘤细胞凋亡的作用及其机制。
方法
2
采用Real-time PCR、Western blot检测成骨细胞及不同骨肉瘤细胞株(SaOS2、MG63、U2OS和HOS)中RanBP9的mRNA和蛋白表达水平。使用慢病毒建立稳定过表达RanBP9的SaOS2骨肉瘤细胞模型,并利用流式细胞仪及TUNEL染色法检测过表达RanBP9组及对照组骨肉瘤细胞凋亡率,使用线粒体膜电位检测过表达RanBP9组及对照组细胞凋亡过程中发生的线粒体膜电位变化。
结果
2
RanBP9在成骨细胞和骨肉瘤细胞株中均呈阳性表达,在成骨细胞中的表达显著高于骨肉瘤细胞(
P
<
0.05),并且在恶性程度较低的骨肉瘤细胞株MG63和HOS中的表达要高于恶性程度较高的细胞株SaOS2和U2OS(
P
<
0.0
5)。过表达RanBP9降低骨肉瘤细胞的增殖能力,促进凋亡率上升(
P
<
0.05)。过表达RanBP9明显增加Caspase-3和Cytc的表达水平,并同时上调Bax/Bcl-2的比率(
P
<
0.05);过表达RanBP9破坏线粒体膜电位,诱导内源性线粒体凋亡信号转导途径。
结论
2
RanBP9通过上调内源性线粒体通路发挥诱导骨肉瘤细胞凋亡的作用。
Objective
2
To investigate the effect of overexpression of Ran-binding protein 9 (RanBP9) on apoptosis of osteosarcoma cells and the underlying mechanism.
Methods
2
Real-time PCR and Western blotting were used to detect the mRNA and protein expression of RanBP9 in cultured osteoblasts and different osteosarcoma cell lines (SaOS2
MG63
U2OS and HOS). In a SaOS2 cell model with stable RanBP9 overexpression induced by transfection with a lentiviral vector
the cell apoptotic rate was determined using flow cytometry and TUNEL assay
and the changes in mitochondrial transmembrane potential were analyzed.
Results
2
RanBP9 was positively expressed in cultured osteoblasts and in the 4 osteosarcoma cell lines. RanBP9 expression was significantly higher in the osteoblasts than in the osteosarcoma cells
and was significantly higher in osteosarcoma cell lines MG63 and HOS than in SaOS2 and U2OS cell lines that had higher levels of malignancy. In SaOS2 cells
overexpression of RanBP9 obviously suppressed cell proliferation and promoted cell apoptosis and significantly increased the expression levels of caspase-3 and Cytc and the ratio of Bax/Bcl-2
resulting also in the dissipation of mitochondrial transmembrane potential and activation of the mitochondrial pathway to induce cell apoptosis.
Conclusion
2
RanBP9 promotes osteosarcoma cell apoptosis
via
the endogenous mitochondrial pathway.
MILLER K D, SIEGEL R L, LIN C C, et al . Cancer treatment and survivorship statistics, 2016[J ] . CA Cancer J Clin, 2016, 66(4): 271-289. DOI: 10.3322/caac.21349.
FEARON E R, VOGELSTEIN B. A genetic model for colorectal tumorigenesis[J]. Cell, 1990, 61(5): 759-767. DOI: 10.1016/0092-8674(90)90186-I.
张樨, 吕杨帆, 戴欢子, 等. RanBP9的表达对骨肉瘤细胞增殖、凋亡及转移的影响[J].第三军医大学学报, 2014, 36(8): 745-749. DOI: 1000-5404(2014)08-0745-05.
ZHANGX, LYU Y F, DAI H Z, et al . Effect of RanBP9 expression on proliferation, apoptosis and metastasis in human osteosarcoma cells[J ] . J Third Mil Med Univ, 2014, 36(8): 745-749. DOI: 1000-5404(2014) 08-0745-05.
YOKOYAMA N, HAYASHI N, SEKI T, et al . A giant nucleopore protein that binds Ran/TC4[J ] . Nature, 1995, 376(6536): 184-188. DOI: 10.1038/376184a0.
MURRIN L C, TALBOT J N. RanBPM, a scaffolding protein in the immune and nervous systems[J]. J Neuroimmune Pharmacol, 2007, 2(3): 290-295. DOI: 10.1007/s11481-007-9079-x.
SALEMI L M, LOUREIRO S O, SCHILD-POULTER C. Characterization of RanBPM molecular determinants that control its subcellular localization[J]. PLoS ONE, 2015, 10(2): e0117655. DOI: 10.1371/journal.pone.0117655.
DAS S, SURESH B, KIM H, et al . RanBPM: A potential therapeutic target for modulating diverse physiological disorders[J ] . Drug Discovery Today, 2017, 22(12). DOI: 10.1016/j.drudis.2017.08.005.
WANG R, WANG H, CARRERA I, et al . COPS5 protein overexpression increases amyloid plaque burden, decreases spinophilin-immunoreactive puncta, and exacerbates learning and memory deficits in the mouse brain[J ] . J Biol Chem, 2015, 290(14): 9299-9309. DOI: 10.1074/jbc.M114.595926.
WANG R, PALAVICINI J P, WANG H, et al . RanBP9 overexpression accelerates loss of dendritic spines in a mouse model of Alzheimer's disease[J ] . Neurobiol Dis, 2014, 69: 169-179. DOI: 10.1016/j.nbd.2014.05.029.
QIN C, ZHANG Q, WU G. RANBP9 suppresses tumor proliferation in colorectal cancer.[J]. Oncol Lett, 2019, 17(5): 4409-4416. DOI: 10.3892/ol.2019.10134.
SHAO S, SUN P H, SATHERLEY L K, et al . Reduced RanBPM expression is associated with distant metastasis in gastric cancer and chemoresistance[J ] . Anticancer Res, 2016, 36(3): 1295-1303.
ZHAO Z, CHENG S, ZABKIEWICZ C, et al . Reduced expression of RanBPM is associated with poorer survival from lung cancer and increased proliferation and invasion of lung cancer cells in vitro [J ] . Anticancer Res, 2017, 37(8): 4389-4397. DOI: 10.21873/anticanres.11833..
WEI J D, JANG J H, KIM J H, et al . RanBPM inhibits BLT2-mediated IL-8 production and invasiveness in aggressive breast cancer cells[J ] . Biochem Biophys Res Commun, 2017, 483(1): 305-311. DOI: 10.1016/j.bbrc.2016.12.147.
DAI H, LV Y F, YAN G N, et al . RanBP9/TSSC3 complex cooperates to suppress anoikis resistance and metastasis via inhibiting Src-mediated Akt signaling in osteosarcoma[J ] . Cell Death Dis, 2016, 7(12): e2572. DOI: 10.1038/cddis.2016.436.
LIN H Y, CHIN Y T, YANG Y C, et al . Thyroid hormone, cancer, and apoptosis[J ] . Compre Physiol, 2016, 6(3): 1221-1237. DOI: 10.1002/cphy.c150035.
BLANDER J M. The many ways tissue phagocytes respond to dying cells[J]. Immunol Rev, 2017, 277(1): 158-173. DOI: 10.1111/imr.12537.
SIDDIQUI W A, AHAD A, AHSAN H. The mystery of BCL2 family: Bcl-2 proteins and apoptosis: an update[J]. Arch Toxicol, 2015, 89(3): 289-317. DOI: 10.1007/s00204-014-1448-7.
GERMAIN M, MATHAI J P, MCBRIDE H M, et al . Endoplasmic reticulum BIK initiates DRP1-regulated remodelling of mitochondrial cristae during apoptosis[J ] . Embo J, 2005, 24(8): 1546-1556. DOI: 10.1038/sj.emboj.7600592.
ZHANG J, MA W, TIAN S, et al . RanBPM interacts with TβRI, TRAF6 and curbs TGF induced nuclear accumulation of TβRI[J ] . Cell Signal, 2014, 26(1): 162-172. DOI: 10.1016/j.cellsig.2013.09.019.
MIKOLAJCZYK M, SHI J, VAILLANCOURT R R, et al . The cyclin-dependent kinase 11p46 isoform interacts with RanBPM[J ] . Biochem Biophys Res Commun, 2003, 310(1): 14-18. DOI: 10.1016/j.bbrc.2003.08.116.
KRAMER S, OZAKI T, MIYAZAKI K, et al . Protein stability and function of p73 are modulated by a physical interaction with RanBPM in mammalian cultured cells[J ] . Oncogene, 2005, 24(5): 938-944. DOI: 10.1038/sj.onc.1208257.
LIU T, ROH S E, WOO J A, et al . Cooperative role of RanBP9 and P73 in mitochondria-mediated apoptosis[J ] . Cell Death Dis, 2013, 4(1): e476. DOI: 10.1038/cddis.2012.203.
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