沉默LIMK1抑制人胃癌BGC823细胞迁移与侵袭

沉默LIMK1抑制人胃癌BGC823细胞迁移与侵袭

杨邦敏,向姝霖,夏红,刘芳,周志刚,苏琦

南华大学肿瘤研究所怀化市第一医院,南华大学肿瘤研究所怀化市第一医院,南华大学肿瘤研究所,南华大学肿瘤研究所,南华大学肿瘤研究所,南华大学肿瘤研究所

收稿日期:2019-06-26 修回日期:2019-06-26 出版日期:2019-07-25
基金资助:
国家自然科学基金（项目编号：81374013；项目名称：二烯丙基二硫上调RORα拮抗Wnt/β-catenin通路抑制人胃癌细胞EMT与迁移侵袭）；国家自然科学基金（项目编号：81102854；项目名称：LIMK1：二烯丙基二硫抑制人胃癌细胞增殖、迁移及侵袭的作用靶点）

Silencing LIMK1 Inhibits Migration and Invasion in Human Gastric Cancer BGC823 Cells

YANG Bangmin,XIANG Shulin,XIA Hong,LIU Fang,ZHOU Zhigang and SU Qi

Cancer Research Institute, University of South China, The Huaihua First hospital,Cancer Research Institute, University of South China, The Huaihua First hospital,Cancer Research Institute, University of South China,Cancer Research Institute, University of South China,Cancer Research Institute, University of South China,Cancer Research Institute, University of South China

Received:2019-06-26 Revised:2019-06-26 Online:2019-07-25

摘要/Abstract

摘要： 目的：探讨沉默LIMK1对人胃癌BGC823细胞迁移与侵袭的影响。方法：RNA干扰技术沉默人胃癌BGC823细胞LIMK1基因，RT-PCR和Western blot检测干扰效率；MTT、细胞划痕和侵袭实验分别检测沉默LIMK1对人胃癌BGC823细胞增殖、迁移与侵袭能力的影响。结果：成功构建沉默LIMK1基因稳定人胃癌BGC823细胞。MTT显示，LIMK1沉默组在24、48、72与96 h较对照组与空载体组的抑制率分别为1.67%、1.45%、0.25%与2.2%和1.60%、1.29%、0.25%与2.2% (P<0.05)。划痕实验显示，12 h和24 h后，沉默组疤痕距离分别为（0.66±0.03）cm与（0.30±0.09）cm，较对照组（0.35±0.03）cm与（0.10±0.03）cm呈时间依赖性降低(P<0.05)。沉默组穿膜细胞数（35±2.86）个，少于对照组（66±3.56）个及空载体组（65±4.35）个 (P<0.05)。结论：沉默LIMK1可抑制人胃癌BGC823细胞增殖、迁移与侵袭。

Abstract: Objective： To investigate the effect of silence LIMK1 on migration and invasion in human gastric cancer BGC823 cells. Methods： RT-PCR and Western blot were used to silence LIMK1 gene in human gastric cancer BGC823 cells.The effects of silence LIMK1 on proliferation， migration and invasion of BGC823 cells were detected by MTT，cell scratch and invasion experiments. Results： Stable LIMK1 gene in human gastric cancer BGC823 cells were successfully constructed. MTT showed that the inhibition rates of the LIMK1 silencing group at 24， 48， 72 and 96h were 1.67%， 1.45%， 0.25% and 2.2%， and 1.60%， 1.29%， 0.25% and 2.2%， respectively， compared with the control group and the empty carrier group (P<0.05).The scratch test showed that after 12h and 24h， the scar distance of the silence group was(0.66±0.03)cm and (0.30±0.09)cm， respectively， which showed a significant time-dependent decrease(P<0.05) compared with that of the control group (0.35±0.03)cm and （0.10±0.03)cm， respectively.The number of transmembrane cells in the silent group was(35±2.86)， which was significantly lower than that in the control group(66±3.56) and the empty carrier group (65±4.35) (P<0.05). Conclusion： Silencing LIMK1 can inhibit proliferation， migration and invasion in human gastric cancer BGC823 cells.

杨邦敏,向姝霖,夏红,刘芳,周志刚,苏琦. 沉默LIMK1抑制人胃癌BGC823细胞迁移与侵袭[J]. .

YANG Bangmin,XIANG Shulin,XIA Hong,LIU Fang,ZHOU Zhigang and SU Qi. Silencing LIMK1 Inhibits Migration and Invasion in Human Gastric Cancer BGC823 Cells[J]. .

参考文献

[1]Bray F,SFerlay J,SSoerjomataram I,Set al. GlobalScancer statisticsS2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries [J]. CASCancerSJ Clin,S2018, 68(6):394-424.
[2]Chen W,SZheng R,SBaade PD,Set al. Cancer statistics in China, 2015 [J]. CA Cancer J Clin, 2016, 66(2):115-32.S
[3]Orditura M,SGalizia G,SSforza V,Set al. Treatment ofSgastric cancer [J]. World J Gastroenterol, 2014, 20(7):1635-49.
[4]Zhang Y,SLi A,SShi J,Set al. ImbalancedSLIMK1Sand LIMK2 expression leads to human colorectal cancer progression and metastasis via promoting β-catenin nuclear translocation [J]. Cell Death Dis,S2018, 9(7):749.
[5]You T,SGao W,SWei J,Set al. Overexpression ofSLIMK1 promotes tumor growth and metastasis in gastricScancer [J]. Biomed Pharmacother, 2015, 69:96-101.
[6]Su J,SZhou Y,SPan Z,Set al. DownregulationSof LIMK1-ADF/cofilin bySDADS inhibits the migrationSandSinvasionSofScolon cancer [J]. Sci Rep, 2017, 7:45624.
[7]Liao Q,SLi R,SZhou R,Set al. LIM kinase 1 interacts with myosin-9 and alpha-actinin-4 and promotes colorectalScancerSprogression [J]. Br JSCancer,S2017, 117(4): 563-71.
[8]Zhao J,SLi D,SFang L. MiR-128-3p suppresses breastScancerScellular progression via targetingSLIMK1 [J]. Biomed Pharmacother,S2019, 115:108947.
[9]Gong H,SZhou L,SKhelfat L,Set al. Rho-Associated Protein Kinase (ROCK) Promotes Proliferation and Migration of PC-3 and DU145 ProstateSCancerSCells by Targeting LIM Kinase 1 (LIMK1) and Matrix Metalloproteinase-2 (MMP-2) [J]. Med Sci Monit,S2019, 25: 3090-9.
[10]Tan Y,SHu H,STan W,Set al. MicroRNA-138 inhibits migration and invasion of non-small cell lungScancerScells by targetingSLIMK1 [J]. Mol Med Rep,S2016, 14(5):4422-8.
[11]Shi B,SMa C,SLiu G,Set al. MiR-106a directly targetsSLIMK1Sto inhibit proliferation and EMT of oral carcinoma cells [J]. Cell Mol Biol Lett,S2019, 24:1.
[12]Yu HX,SWang XL,SZhang LN,Set al. MicroRNA-384 inhibits the progression of esophageal squamous cell carcinoma through blockade of the LIMK1/cofilin signaling pathway by binding toSLIMK1 [J]. Biomed Pharmacother, 2019, 109:751-61.
[13]Zhang HY,SZhang J,SHao CZ,Set al. LC-0882 targets PAK4 and inhibits PAK4-related signaling pathways to suppress the proliferation and invasion of gastricScancerScells [J]. Am J Transl Res,S2017, 9(6): 2736-47.
[14]Lu H,SChen J,SLuo Y,Set al. Curcolonol suppresses the motility of breastScancerScells by inhibiting LIM kinaseS1 to downregulate cofilin 1 phosphorylation [J]. Int J Oncol, 2018, 53(6): 2695-704.
[15]Charles MD,SBrookfield JL,SEkwuru TC,Set al. Discovery, Development, and SAR of Aminothiazoles as LIMK Inhibitors with Cellular Anti-Invasive Properties [J]. J Med Chem, 2015, 58(20):8309-13.
[16]Guo B,SZhang T,SSu J,Set al. Oxymatrine targets EGFR(p-Tyr845) and inhibits EGFR-related signaling pathways to suppress the proliferation and invasion of gastricScancerScells [J]. CancerSChemother Pharmacol,S2015,75(2):353-63.
[17]Fu J,SYu J,SChen J,Set al. In vitro inhibitory properties of sesquiterpenes from Chloranthus serratus on cell motility via down-regulation ofSLIMK1Sactivation in human breastScancer [J]. Phytomedicine,S2018, 49:23-31.
[18]Zhou Y, Su J, Shi L, et al. DADS downregulates the Rac1-ROCK1/PAK1-LIMK1- ADF/cofilin signaling pathway, inhibiting cell migration and invasion [J]. Oncol Rep, 2013, 29(2):605-612.
[19]Su B, He H, Wang L, et al. Chk1, but not Chk2, is responsible for G2/M phase arrest induced by diallyl disulfide in human gastric cancer BGC823 cells. Food Chem Toxicol,S2014, 68:61-70.
[20]Su B, Su J, He H, et al. Identification of potential targets for diallyl disulfide in human gastric cancer MGC-803 cells using proteomics approaches. Oncol Rep,S2015, 33(5):2484-2494.
[21]Su B,SSu J,SZeng Y,Set al. Diallyl disulfide inhibits TGF-β1-induced upregulation of Rac1 and β-catenin in epithelial-mesenchymal transition and tumor growth of gastric cancer [J]. Oncol Rep,S2018, 39(6):2797-806.

快速检索引用检索图表检索高级检索