[1]刘军言,刘佳佳,吴益西,等.敲低肌切蛋白可抑制胃肠道间质瘤细胞的增殖能力[J].第三军医大学学报,2016,38(12):1422-1426.
 Liu Junyan,Liu Jiajia,Wu Yixi,et al.Knockdown of scinderin inhibits proliferation in gastrointestinal stromal tumor cells[J].J Third Mil Med Univ,2016,38(12):1422-1426.
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
38卷
期数:
2016年第12期
页码:
1422-1426
栏目:
基础医学
出版日期:
2016-06-30

文章信息/Info

Title:
Knockdown of scinderin inhibits proliferation in gastrointestinal stromal tumor cells
作者:
刘军言刘佳佳吴益西陈骞林勇钱锋
第三军医大学西南医院:全军普通外科中心,微创胃肠外科中心,全军临床病理学研究所
Author(s):
Liu Junyan Liu Jiajia Wu Yixi Chen Qian Lin Yong Qian Feng

Center of General Surgery, Center of Minimal Invasive Gastrointestinal Surgery, Institute of Pathology, Southwest Hospital, Third Military Medical University, Chongqing, 400038, China

关键词:
肌切蛋白胃肠道间质瘤增殖伊马替尼
Keywords:
scinderin gastrointestinal stromal tumor proliferation imatinib
分类号:
R394.2; R730.24; R735
文献标志码:
A
摘要:

目的      探讨肌切蛋白(scinderin,SCIN)对胃肠道间质瘤(gastrointestinal stromal tumor,GIST)细胞增殖能力的影响。      方法      用shRNA慢病毒干扰技术将SCIN在GIST细胞中稳定敲低,并通过Western blot和qRT-PCR检测敲低效率。通过CCK-8实验检测伊马替尼IC50和SCIN对GIST细胞增殖能力的影响,通过平板克隆形成实验检测SCIN对GIST细胞克隆形成能力的影响,通过裸鼠皮下成瘤实验检测SCIN对移植瘤生长的影响。实验按照对GIST细胞进行的不同处理分组:WT组(野生型组,未进行转染),Mock组(阴性对照组,转染阴性对照序列),shSCIN 1组(转染SCIN干扰序列1),shSCIN 2组(转染SCIN干扰序列2)。      结果      shSCIN 1组和shSCIN 2组GIST细胞在增殖能力上明显低于Mock组,增殖能力在第2~7天均有明显差异(n=4,P<0.05)。移植瘤实验中shSCIN 1组较Mock组的移植瘤质量显著降低:shSCIN 1组vs Mock组=(0.175 0±0.045 6)g vs (0.334 1±0.016 5)g(n=4,P<0.01)。平板克隆形成实验显示shSCIN 1组克隆形成数显著少于Mock组,分别为59.67±5.68和123.33±2.52(n=3,P<0.01)。在使用0.57 μmol/L的伊马替尼处理WT组细胞1、3、5 d后,SCIN的表达随处理时间而降低。      结论      敲低SCIN可以抑制GIST细胞在体外和体内环境中的增殖能力,伊马替尼可以抑制GIST细胞中SCIN的表达,SCIN可能成为GIST的治疗靶点。

Abstract:

Objective      To determine the role of scinderin (SCIN) in regulating proliferation of gastrointestinal stromal tumor (GIST) cells.       Methods      SCIN-knockdown GIST cells were established with SCIN-specific shRNAs (shSCINs). The knockdown efficiency was determined by Western blotting and qRT-PCR. CCK-8 assay was used to evaluate imatinib IC50 and the effect of shSCIN on proliferation. Plate colony formation assay was used to test the effect of shSCINs on colony formation of GIST cells. Xenograft assay in nude mice was employed to test the effect of shSCINs on tumor growth. According to different treatment, GIST cells were divided into 4 groups: wild type group (WT group, no transfection), mock group (transfected with scrambled siRNA), shSCIN1 group (transfected with shSCIN1) and shSCIN 2 group (transfected with shSCIN2).       Results      The cell proliferation of shSCIN1 group and shSCIN2 group was significantly decreased compared with the mock group. The proliferation difference was significant from the second day to the seventh day (n=4, P<0.05). Xenografts derived from shSCIN1 cells had significantly lower tumor weight than those derived from mock cells (0.1750±0.0456 vs 0.3341±0.0165 g, n=4, P<0.01). The colony numbers derived from shSCIN1 cells and mock cells were 59.67±5.68 and 123.33±2.52, respectively (n=3, P<0.01). After treatment with 0.57 μmol/L imatinib in WT cells for 1, 3 and 5 d, the expression of SCIN was found to be decreased in a time-dependent manner.       Conclusion      Knockdown of SCIN inhibits the proliferation of GIST cells both in vitro and in vivo. Imatinib inhibits the expression of SCIN. SCIN may be a promising therapeutic target for GIST.

参考文献/References:

[1]Kanda T. Criminal or bystander: imatinib and second primary malignancy in GIST patients[J]. Chin J Cancer Res, 2013, 25(5): 490-492. DOI: 10.3978/j.issn.1000-9604.2013.10.15
[2]Wang C, Jin M S, Zou Y B, et al. Diagnostic significance of DOG-1 and PKC-theta expression and c-Kit/PDGFRA mutations in gastrointestinal stromal tumours[J]. Scand J Gastroenterol, 2013, 48(9): 1055-1065. DOI: 10.3109/00365521.2013.816770
[3]Muhlenberg T, Grunewald S, Treckmann J, et al. Inhibition of KIT-glycosylation by 2-deoxyglucose abrogates KIT-signaling and combination with ABT-263 synergistically induces apoptosis in gastrointestinal stromal tumor[J]. PLoS One, 2015, 10(3): e0120531. DOI: 10.1371/journal.pone.0120531
[4]Nag S, Larsson M, Robinson R C, et al. Gelsolin: the tail of a molecular gymnast[J]. Cytoskeleton (Hoboken), 2013, 70(7): 360-384. DOI: 10.1002/cm.21117
[5]Marcu M G, Zhang L, Elzagallaai A, et al. Localization by segmental deletion analysis and functional characterization of a third actin-binding site in domain 5 of scinderin[J]. J Biol Chem, 1998, 273(6): 3661-3668.
[6]Miura N, Takemori N, Kikugawa T, et al. Adseverin: a novel cisplatin-resistant marker in the human bladder cancer cell line HT1376 identified by quantitative proteomic analysis[J]. Mol Oncol, 2012, 6(3): 311-322. DOI: 10.1016/j.molonc.2011.12.002
 [7]Liu H, Shi D, Liu T, et al. Lentivirus-mediated silencing of SCIN inhibits proliferation of human lung carcinoma cells[J]. Gene, 2015, 554(1): 32-39. DOI: 10.1016/j.gene.2014.10.013
[8]Wang D, Sun S Q, Yu Y H, et al. Suppression of SCIN inhibits human prostate cancer cell proliferation and induces G0/G1 phase arrest[J]. Int J Oncol, 2014, 44(1): 161-166. DOI: 10.3892/ijo.2013.2170
[9]Fukuda K, Saikawa Y, Sako H, et al. Establishment and characterization of novel cell lines and xenografts from patients with gastrointestinal stromal tumors[J]. Oncol Rep, 2013, 30(1): 71-78. DOI: 10.3892/or.2013.2425
[10]Datar M, Khanna R. Inpatient burden of gastrointestinal stromal tumors in the United States[J]. J Gastrointest Oncol, 2012, 3(4): 335-341. DOI: 10.3978/j.issn.2078-6891.2012.037
[11]Hayashi Y, Bardsley M R, Toyomasu Y, et al. Platelet-Derived Growth Factor Receptor-alpha Regulates Proliferation of Gastrointestinal Stromal Tumor Cells With Mutations in KIT by Stabilizing ETV1[J]. Gastroenterology, 2015, 149(2): 420-432.e16. DOI: 10.1053/j.gastro.2015.04.006
[12]Xu C W, Lin S, Wang W L, et al. Analysis of mutation of the c-Kit gene and PDGFRA in gastrointestinal stromal tumors[J]. Exp Ther Med, 2015, 10(3): 1045-1051. DOI: 10.3892/etm.2015.2613
[13]O’Brien K M, Orlow I, Antonescu C R, et al. Gastrointestinal stromal tumors, somatic mutations and candidate genetic risk variants[J]. PLoS One, 2013, 8(4): e62119. DOI: 10.1371/journal.pone.0062119
[14]Tan C J, Yang J L, Crowe P, et al. Targeted therapy in soft tissue sarcoma-a novel direction in therapeutics[J]. Chin Clin Oncol, 2013, 2(3): 22. DOI: 10.3978/j.issn.2304-3865.2013.08.01
[15]Yuzawa S, Opatowsky Y, Zhang Z, et al. Structural basis for activation of the receptor tyrosine kinase KIT by stem cell factor[J]. Cell, 2007, 130(2): 323-334. DOI: 10.1016/j.cell.2007.05.055
[16]Wang J, Cai J, Huang Y, et al. Nestin regulates proliferation and invasion of gastrointestinal stromal tumor cells by altering mitochondrial dynamics[J]. Oncogene, 2015. DOI: 10.1038/onc.2015.370 [Epub ahead of print]
[17]Lin P, Sun X, Feng T, et al. ADAM17 regulates prostate cancer cell proliferation through mediating cell cycle progression by EGFR/PI3K/AKT pathway[J]. Mol Cell Biochem, 2012, 359(1/2): 235-243. DOI: 10.1007/s11010-011-1018-8
[18]Li J, Dang Y, Gao J, et al. PI3K/AKT/mTOR pathway is activated after imatinib secondary resistance in gastrointestinal stromal tumors (GISTs)[J]. Med Oncol, 2015, 32(4): 111. DOI: 10.1007/s12032-015-0554-6
[19]Ma Y Y, Yu S, He X J, et al. Involvement of c-KIT mutation in the development of gastrointestinal stromal tumors through proliferation promotion and apoptosis inhibition[J]. Onco Targets Ther, 2014, 7: 637-643. DOI: 10.2147/OTT.S60458
[20]李健.胃肠道间质瘤的分子靶向治疗[J].中华消化外科杂志,2013,12(4): 253-256.DOI:10.3760/cma.j.issn.1673-9752.2013.04.004

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更新日期/Last Update: 2016-06-07