RNA干扰YWHAZ基因对结肠癌HCT-8细胞增殖的影响

doi:10.13481/j.1671-587x.20150220

吉林大学学报(医学版) ›› 2015, Vol. 41 ›› Issue (02): 307-310.doi: 10.13481/j.1671-587x.20150220

RNA干扰YWHAZ基因对结肠癌HCT-8细胞增殖的影响

张成才¹, 宋艳艳², 周梦娇¹, 崔志倩¹, 张西臣³, 邢沈阳¹

1. 吉林大学动物科学学院动物科学系, 吉林长春 130062;
2. 吉林大学第二医院肾病内科, 吉林长春 130041;
3. 吉林大学动物医学学院预防兽医学系, 吉林长春 130062

收稿日期:2014-11-06 出版日期:2015-03-28 发布日期:2015-04-04
通讯作者: 邢沈阳, 教授, 硕士研究生导师(Tel:0431-87835138, E-mail:xingsy6177@163.com) E-mail:xingsy6177@163.com
作者简介:张成才(1989-), 男, 山东省潍坊市人, 在读动物科学硕士, 主要从事肿瘤遗传学方面的研究。
基金资助:
吉林省科技厅科技发展计划项目资助课题(2003055025,20106044);吉林省自然科学基金资助课题(20101585)

Effect of RNA interference of YWHAZ gene on proliferation of colon cancer HCT-8 cells

ZHANG Chengcai¹, SONG Yanyan², ZHOU Mengjiao¹, CUI Zhiqian¹, ZHANG Xichen³, XING Shenyang¹

1. Department of Animal Science, School of Animal Science, Jilin University, Changchun 130062, China;
2. Department of Nephrology, Second Hospital, Jilin University, Changchun 130041, China;
3. Department of Preventive Veterinary Medicine, School of Veterinary Medicine, Jilin University, Changchun 130062, China

Received:2014-11-06 Online:2015-03-28 Published:2015-04-04

摘要/Abstract

摘要：

目的:探讨中RNA干扰YWHAZ基因对结肠癌HCT-8细胞增殖的影响,阐明其作用机制。方法:构建靶向基因YWHAZ的siRNA载体重组质粒pGPU6/GFP/Neo-YWHAZ-792、pGPU6/GFP/Neo-YWHAZ-733、pGPU6/GFP/Neo-YWHAZ-612和pGPU6/GFP/Neo-YWHAZ-505作为RNA干扰质粒组,同时设立阴性对照质粒组(pGPU6/GFP/Neo-shNC质粒);采用荧光定量PCR检测YWHAZ mRNA的表达水平和表达抑制率;采用MTT法检测HCT-8细胞的增殖率。结果:倒置荧光显微镜下观察,细胞转染率为60%。荧光定量PCR检测,RNA干扰组YWHAZ mRNA表达水平低于阴性对照组(P<0.01),平均抑制率为52.86%。MTT检测,与阴性对照组比较,RNA干扰质粒组细胞增殖率明显降低(P<0.01),平均细胞增殖率为68.75%。结论:干扰YWHAZ可抑制该基因的转录,并抑制HCT-8细胞的增殖。

关键词: YWHAZ基因, 结肠肿瘤, HCT-8细胞, RNA干扰

Abstract:

Objective To investigate the effect of RNA interference of YWHAZ gene on the cell proliferation of colon cancer HCT-8 cells,and to elucidate the mechanism.Methods Four siRNA plasmids targeting human YWHAZ gene,including pGPU6/GFP/Neo-YWHAZ-792,pGPU6/GFP/Neo-YWHAZ-733,pGPU6/GFP/Neo-YWHAZ-612,and pGPU6/GFP/Neo-YWHAZ-505, were constructed and regarded as RNA interference plasmid groups. pGPU6/GFP/Neo-shNC plasmid was regarded as negative control plasmid group. RT-PCR was used to detect the expression of YWHAZ mRNA and the inhibitory rates of HCT-8 cells; the proliferation rate of HCT-8 cells was detected by MTT method.Results The transfection rate of cells was 60% under inverted fluorescence microscope;the RT-PCR results showed that the levels of YWHAZ mRNA in RNA interference plasmid groups were lower than those in negative control plasmid group(P<0.01),and the average inhibitory rate was 52.86%. The MTT results showed that compared with negative control plasmid group,the proliferation rates of HCT-8 cells in RNA interference plasmid groups were decreased (P<0.01),and the average proliferation rate was 68.75%.Conclusion Interference of YWHAZ gene might inhibit the transcription of the gene,and further inhibit the proliferation of HCT-8 cells.

Key words: YWHAZ gene, colon neoplasms, HCT-8 cells, RNA interference

中图分类号:

R735.3

张成才, 宋艳艳, 周梦娇, 崔志倩, 张西臣, 邢沈阳. RNA干扰YWHAZ基因对结肠癌HCT-8细胞增殖的影响[J]. 吉林大学学报(医学版), 2015, 41(02): 307-310.

ZHANG Chengcai, SONG Yanyan, ZHOU Mengjiao, CUI Zhiqian, ZHANG Xichen, XING Shenyang. Effect of RNA interference of YWHAZ gene on proliferation of colon cancer HCT-8 cells[J]. Journal of Jilin University Medicine Edition, 2015, 41(02): 307-310.

参考文献

[1] Feng E,Chen H,Li Ya,et al. Gene cloning,expression,and function analysis of SpL 14-3-3ζ in Spodptera litura and its response to the entomopathogenic fungus Nomuraea rileyi [J]. Comp Biochem Physiol,Part B,2014,172-173:49-56.

[2] Chen MJ,Liu TF,Xu LN,et al. Direct interaction of 14-3-3ζ with ezrin promotes cell migration by regulating the formation of membrane ruffle [J]. J Mol Biol,2014,426(18):3118-3133.

[3] Gao XJ,He YJ,Gao LM,et al. Ser9-phosphorylated GSK3β induced by 14-3-3ζ actively antagonizes cell apoptosis in a NF-κB dependent manner [J]. Biochem Cell Biol,2014,92(5):1-8.

[4] Zhang Z,Luo X,Ding S,et al. MicroRNA-451 regulates p38 MAPK signaling by targeting of Ywhaz and suppresses the mesangial hypertrophy in early diabetic nephropathy [J]. FEBS Lett,2012,586 (1):20-26.

[5] 臧东钰. 14-3-3zeta在T1期非小细胞肺癌中的表达及与肺癌侵袭、转移和凋亡关系的研究 [D]. 沈阳:中国医科大学,2010.

[6] Shikano S,Coblitz B,Wu M,et al. 14-3-3 proteins:regulation of endoplasmic reticulum localization and surface expression of membrane proteins [J]. Trends Cell Biol,2006,16 (7):370-375.

[7] 张慧丰,左萍萍. 肺癌转移机制研究进展 [J]. 医学研究进展,2013,10(42):4-6.

[8] Liu M,Liu XX,Ren PF,et al. A cancer-related protein 14-3-3ζ is a potential tumor-associated antigen in immunodiagnosis of hepatocellular carcinoma [J]. Tumor Biol,2014,35(5):4247-4256.

[9] 巴艳华,李惠平,赵红梅,等. mdm2/p53通路相关的乳腺癌差异表达基因分析 [J]. 癌症进展,2008,6(2):110-116.

[10] 王雅雯. YWHAZ基因过表达载体的构建及对肿瘤迁移能力的影响 [D]. 长春:吉林大学,2013.

[11] Larriba MJ,Casado-Vela J,Pendás-Franco N, et al. Novel snail1 target proteins in human colon cancer identified by proteomic analysis [J]. PLoS One,2014,5(4):e10221.

[12] 杨建军,秦环龙. 14-3-3蛋白与肿瘤发生发展的研究进展[J]. 世界华人消化杂志,2010,18(28):2997-3002.

[13] Chaithirayanon K,Grams R,Vichasri-Grams S,et al. Molecular and immunological characterization of eneoding gene and 14-3-3 protein 1 in Fasciola gigantica [J]. Parasitology,2006,133(6):763-775.

[14] Pan YF,Qin T,Feng L,et al. Expression profile of altered long non-coding RNAs in patients with HBV-associated hepatocellular carcinoma [J]. J Huazhong Univ Sci Technol,2013,33(1):96-101.

[15] Bajpai U,Sharma R,Kausar T,et al. Clinical significance of 14-3-3 zeta in human esophageal cancer [J]. Int J Biol Markers,2008,23 (4):231-237.

[16] Tsukamoto Y,Nakada C,Noguchi T,et al. MicroRNA-375 is downregulated in gastric carcinomas and regulates cell survival by targeting PDK1 and 14-3-3 zeta [J]. Cancer Res,2010,70(6):2339-2349.

[17] Maxwell SA,Li ZG,Jaye D,et al. 14-3-3 zeta mediates resistance of diffuse large B cell lymphoma to an anthracycline-based chemotherapeutic regimen [J]. J Biol Chem,2009,284(33):22379-33389.

[18] 袁亚丽. 靶向14-3-3ζ基因siRNA的重组腺相关病毒载体的构建与表达 [D]. 广州:华南理工大学,2011.

[19] Li Z,Zhao J,Du Y,et al. Down-regulation of 14-3-3ζ suppresses anchorage-independent growth of lung cancer cells through anoikis activation [J].Proc Nati Acad Sci,2008,105(1):162-167.

[20] Matta A,Bahadur S,Duggal R,et al. Over-expression of 14-3-3zeta is an early event in oral cancer [J]. BMC Cancer,2007,7:169-180.

[21] 王鑫,陈玲,孙飞,等.CXCR7-shRNA靶向抑制结肠癌SW620细胞的体外实验研究[J].西安交通大学学报:医学版,2014,35(4):460-464,480.

[22] 田筱青,孙丹凤,房静远.GSTM1基因多态性及启动子甲基化在结肠癌细胞系中的研究[J].西安交通大学学报:医学版,2014,35(1):56-58,93.

RNA干扰YWHAZ基因对结肠癌HCT-8细胞增殖的影响

Effect of RNA interference of YWHAZ gene on proliferation of colon cancer HCT-8 cells

RichHTML

PDF (PC)

赞

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

Metrics

本文评价

推荐阅读 0

[1]	王廷刚, 薛峰, 李宇, 牛兆健, 王野. miR-26a靶向HMGA1基因对结肠癌细胞生长、侵袭和迁移的影响[J]. 吉林大学学报(医学版), 2018, 44(06): 1205-1211.
[2]	周刘祥, 林琳, 胡鑫敏, 王晓春. 内分泌腺性血管内皮生长因子对结肠癌LoVo细胞增殖和迁移的促进作用[J]. 吉林大学学报(医学版), 2018, 44(03): 516-520.
[3]	王玥, 刘畅, 朴仙姬, 张冬云, 孟令旗, 王浩, 王加茹, 罗英花, 孙虎男, 金成浩. 四溴苯三唑对人结肠癌SW480细胞凋亡的诱导作用及其机制[J]. 吉林大学学报(医学版), 2017, 43(06): 1148-1154.
[4]	曹艳莎, 李婳, 陈明红, 刘宝琴, 王华芹, 李宁. 组织芯片技术检测BAG3蛋白在结肠癌组织中的表达及其临床意义[J]. 吉林大学学报(医学版), 2017, 43(06): 1177-1181.
[5]	王波, 沈倩倩, 张华, 冯智英. 羟考酮联合右美托咪定用于腹腔镜下结肠癌根治术术后镇痛效果评价[J]. 吉林大学学报(医学版), 2017, 43(06): 1231-1236.
[6]	石俊忠, 庄建彬, 张传山, 刘贤明, 孔大陆. β-tubulin Ⅲ在结肠腺癌组织中的表达及其临床意义[J]. 吉林大学学报(医学版), 2017, 43(04): 770-775.
[7]	张晶, 王志成, 赵大力, 卢晓倩, 沈智渊, 齐亚莉. 电离辐射对肺癌H460细胞凋亡的影响及其机制[J]. 吉林大学学报(医学版), 2017, 43(03): 522-526.
[8]	黄志立, 张丽君, 伍丽华, 王妍, 杨汝德. RNA干扰血管紧张素原基因表达对前脂肪细胞3T3-L1分化的影响[J]. 吉林大学学报(医学版), 2015, 41(06): 1195-1200.
[9]	姜恩平, 李贺, 于春艳, 朱伟. 五味子乙素通过p38MAPK信号通路对结肠癌SW480细胞凋亡和侵袭的影响[J]. 吉林大学学报(医学版), 2015, 41(04): 675-679.
[10]	陈伟莉, 张晨, 张旭浩, 刘冬影, 赵玺. 紫薯花青素抗结肠癌作用及其诱导细胞凋亡机制[J]. 吉林大学学报(医学版), 2015, 41(04): 785-789.
[11]	方芳, 张万生, 唐化勇, 赵良中, 陈爽, 李强, 王立国. CD147 RNA干扰对前列腺癌LNCaP-AI细胞体外侵袭力的影响[J]. 吉林大学学报(医学版), 2015, 41(03): 460-463.
[12]	王珂, 邹积艳, 汪丽, 杜珍武, 王俊容, 叶聪, 潘颖. RhoC慢病毒干扰载体的构建及其对卵巢癌SKOV3细胞中RhoC mRNA表达的影响[J]. 吉林大学学报(医学版), 2015, 41(02): 304-306.
[13]	刘畅, 高薇, 冯欣. RNA干扰沉默STAT3对胶原诱导大鼠类风湿性关节炎的治疗作用[J]. 吉林大学学报(医学版), 2015, 41(01): 77-82.
[14]	刘晓康, 韩欣冶, 林英嘉, 胡方舟, 李杨, 李清. 桦木醇对人结肠癌SW480细胞增殖和凋亡的影响[J]. 吉林大学学报(医学版), 2015, 41(01): 39-43.
[15]	蒋清虎，罗伟，温路，胡惠雯，邓大炜，吴忠均. ARK5在肝癌组织中的表达及其对肝癌SMMC-7721细胞生长的影响[J]. 吉林大学学报(医学版), 2014, 40(04): 743-747.