沉默ANK2基因表达对胃癌细胞增殖、周期以及转移能力的影响
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摘要
目的探究沉默细胞膜锚蛋白(ANK2)基因表达对人胃癌细胞SGC-7901增殖、侵袭、细胞周期的影响及作用机制。方法实验分为对照组、无义组、siRNA-ANK2组,胃癌细胞SGC-7901转染siRNA-ANK2或siRNA无义序列,蛋白质印迹法(Western blotting)检测细胞中ANK2蛋白的水平。CCK-8法检测细胞的活性,流式细胞术检测细胞周期变化,Transwell法检测细胞的侵袭力。结果 SGC-7901细胞转染siRNA-ANK2后,与对照组相比,siRNA-ANK2组ANK2蛋白表达量显著降低(P<0.05),细胞活性明显降低(P<0.05);siRNA-ANK2组细胞周期G0/G1期较对照组显著升高,S期降低(P<0.05),侵袭数目明显降低(P<0.05)。结论沉默ANK2基因抑制胃癌细胞SGC-7901增殖和转移,阻碍细胞周期G1向S期转换。
Objective To investigate the effect of silencing cell membrane ankyrin B(ANK2) gene expression on the proliferation,invasion and cell cycle of human gastric cancer cell line SGC-7901 and its mechanism. Methods The experiments were divided into control group,nonsense group and siRNA-ANK2 group. Gastric cancer cell SGC-7901 was transfected into siRNA-ANK2 or siRNA nonsense sequence. The expression of ANK2 protein was detected by Western blotting. The cell activity was detected by CCK-8 assay. Flow cytometry was used to detect cell cycle changes. Transwell method was used to detect cell invasiveness. Results Compared with the control group,the expression of ANK2 protein in siRNA-ANK2 group was decreased significantly after transfected into siRNA-ANK2( P < 0. 05),cell activity was decreased significantly( P < 0. 05). The G0/G1 phase of siRNA-ANK2 group was significantly higher than that of the control group,S phase was decreased( P < 0. 05),and the number of invasion was significantly decreased( P <0. 05). Conclusion Silencing ANK2 gene can inhibit the proliferation,induce metastasis of gastric cancer cell line SGC-7901,and hinder the transition from cell cycle G1 to S phase.
Objective To investigate the effect of silencing cell membrane ankyrin B(ANK2) gene expression on the proliferation,invasion and cell cycle of human gastric cancer cell line SGC-7901 and its mechanism. Methods The experiments were divided into control group,nonsense group and siRNA-ANK2 group. Gastric cancer cell SGC-7901 was transfected into siRNA-ANK2 or siRNA nonsense sequence. The expression of ANK2 protein was detected by Western blotting. The cell activity was detected by CCK-8 assay. Flow cytometry was used to detect cell cycle changes. Transwell method was used to detect cell invasiveness. Results Compared with the control group,the expression of ANK2 protein in siRNA-ANK2 group was decreased significantly after transfected into siRNA-ANK2( P < 0. 05),cell activity was decreased significantly( P < 0. 05). The G0/G1 phase of siRNA-ANK2 group was significantly higher than that of the control group,S phase was decreased( P < 0. 05),and the number of invasion was significantly decreased( P <0. 05). Conclusion Silencing ANK2 gene can inhibit the proliferation,induce metastasis of gastric cancer cell line SGC-7901,and hinder the transition from cell cycle G1 to S phase.
引文
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[2]SHMULEVICH I.Large-scale molecular characterization and analysis of gastric cancer[J].Chin J Cancer,2014,33(8):369-370.
[3]SIEGEL R L,MILLER K D,JEMAL A.Cancer statistics,2017[J].CA Cancer J Clin,2017,67(1):7-30.DOI:10.3322/caac.21387.
[4]CHEN W,ZHENG R,BAADE P D,et al.Cancer statistics in China,2015[J].CA Cancer J Clin,2016,66(2):115-132.DOI:10.3322/caac.21338.
[5]王思萌,高柳村,帖君,等.Cetuximab抑制胃癌SGC7901/ADR细胞的增殖并增加其化疗敏感性[J].现代生物医学进展,2013,13(12):2253-2256,2264.WANG S M,GAO L C,TIE J,et al.In vitro proliferation repression and an increase of chemo-sensitivity of Cetuximab as monotherapy in chemo-refractory gastric cancer cell line SGC7901/ADR[J].Progress in Modern Biomedicine,2013,13(12):2253-2256,2264.
[6]KHL S J,KHL M.On the role of Wnt/β-catenin signaling in stem cells[J].Biochim Biophys Acta,2013,1830(2):2297-2306.DOI:10.1016/j.bbagen.2012.08.010
[7]WNG T,ABOU-OUF H,HEGAZY S A,et al.Ankyrin G expression is associated with androgen receptor stability,invasiveness,and lethal outcome in prostate cancer patients[J].J Mol Med(berl),2016,94(12):1411-1422.DOI:10.1007/s00109-016-1458-4.
[8]罗静.RNA干扰在抗肿瘤中的作用研究进展[J].新乡医学院学报,2017,34(4):340-343.DOI:10.7683.xxyxyxb.2017.04.025.
[9]ZHANG G,WANG Z,QIAN F,et al.Silencing of the ABCC4 gene by RNA interference reverses multidrug resistance in human gastric cancer[J].Oncol Rep,2015,33(3):1147-1154.DOI:10.3892/or.2014.3702.
[10]曹大龙,刘黎明.RNA干扰技术在肿瘤基因治疗中的研究进展[J].医学综述,2015,21(14):2566-2569.DOI:10.3969/j.issn.1006-2084.2015.14.026.CAO D L,LIU L M.Research progress of RNA interference in tumor gene therapy[J].Medical Recapitulate,2015,21(14):2566-2569.DOI:10.3969/j.issn.1006-2084.2015.14.026.
[11]JIANG C,CHEN X,ALATTRAR M,et al.MicroRNAs in tumorigenesis,metastasis,diagnosis and prognosis of gastric cancer[J].Cancer Gene Ther,2015,22(6):291-301.DOI:10.1038/cgt.2015.19.
[12]LIANG L,FANG J Y,XU J.Gastric cancer and gene copy number variation:emerging cancer drivers for targeted therapy[J].Oncogene,2016,35(12):1475-1482.DOI:10.1038/onc.2015.209.
[13]NISHIDA T,EGASHIRA Y,AKUTAGAWA H,et al.Predictors of lymph node metastasis in T1 colorectal carcinoma:an immunophenotypic analysis of 265 patients[J].Dis Colon Rectum,2014,57(8):905-915.DOI:10.1097/DCR.0000000000000168.
[14]DAZ-LPEZ A,MORENO-BUENO G,CANO A.Role of microRNA in epithelial to mesenchymal transition and metastasis and clinical perspectives[J].Cancer Manag Res,2014,25(6):205-216.DOI:10.2147/CMAR.S38156.
[15]CAO W,WEI W,ZHAN Z,et al.Role of miR-647 in human gastric cancer suppression[J].Oncol Rep,2017,37(3):1401-1411.
[16]BABINA I S,MCSHERRY E A,DONATELLO S,et al.A novel mechanism of regulating breast cancer cell migration via palmitoylation-dependent alterations in the lipid raft affiliation of CD44[J].Breast Cancer Res,2014,16(1):R19.DOI:10.1186/bcr3614.
[17]CATTARUZZA F,JOHNSON C,LEGGIT A,et al.Transient receptor potential ankyrin 1 mediates chronic pancreatitis pain in mice[J].Am J Physiol Gastrointest Liver Physiol,2013,304(11):G1002-G1012.DOI:10.1152/ajpgi.00005.2013.
[18]MONTANINI L,LASAGNA L,BARILI V,et al.MicroRNA cloning and sequencing in osteosarcoma cell lines:differential role of miR-93[J].Cell Oncol(Dordr),2012,35(1):29-41.DOI:10.1007/s13402-011-0059-z.