[1]高红伟),李印),郭长青)#,等.腺病毒介导的干扰素β基因对SMMC7721细胞凋亡的影响*[J].郑州大学学报(医学版),2009,(06):1178-1181.
 GAO Hongwei),LI Yin),GUO Changqing),et al.Effects of adenovirusmediated interferonβ gene on apoptosis of SMMC7721 cells[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2009,(06):1178-1181.
点击复制

腺病毒介导的干扰素β基因对SMMC7721细胞凋亡的影响*()
分享到:

《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2009年06期
页码:
1178-1181
栏目:
论著
出版日期:
2009-11-01

文章信息/Info

Title:
Effects of adenovirusmediated interferonβ gene on apoptosis of SMMC7721 cells
作者:
高红伟1)李印2)郭长青1)#马军3)张丽荣1)曹新广2)于泳3)
1)郑州大学第一附属医院消化内科 郑州 4500522)河南省肿瘤医院普外科 郑州 4500033)郑州大学第二附属医院消化内科 郑州 450014
Author(s):
GAO Hongwei1)LI Yin2)GUO Changqing1)MA Jun3)ZHANG Lirong1)CAO Xinguang2)YU Yong3)
1)Department of Gastroenterology,the First Affiliated Hospital,Zhengzhou University, Zhengzhou 450052 2)Department of General Surgery,Tumor Hospital of Henan Province,Zhengzhou 4500033)Department of Gastroenterology,the Second Affiliated Hospital,Zhengzhou Univercity,Zhengzhou 450014
关键词:
腺病毒载体干扰素β肝癌凋亡SMMC7721细胞
Keywords:
adenovirus vectorinterferonβhepatocellular carcinomaapoptosisSMMC7721 cell
分类号:
R735.7
摘要:
观察腺病毒介导的干扰素β(IFNβ)基因能否诱导人肝癌SMMC7721细胞体外凋亡。方法:用HEK293A细胞扩增重组腺病毒介导的人干扰素β(AdhIFNβ)基因,经纯化后用空斑形成实验法测定病毒滴度;分别用AdhIFNβ基因和重组腺病毒介导的绿色荧光蛋白(AdGFP)基因转染人肝癌SMMC7721细胞,并设空白对照组,应用免疫细胞化学方法检测hIFNβ基因的表达,应用Hochest 33342荧光染色法和流式细胞术检测转染重组AdhIFNβ基因组、空载体组和对照组细胞的凋亡情况。结果:重组腺病毒经HEK293A细胞扩增、纯化后滴度可达2×1011pfu/mL,且40 MOI的腺病毒即可获得95%以上的转染效率;免疫细胞化学法检测到外源性hIFNβ基因在SMMC7721细胞中的蛋白表达产物;荧光染色法检测到重组AdhIFNβ基因组细胞明显发生了凋亡,而重组AdGFP组和空白对照组细胞凋亡不明显;流式细胞术检测到转染重组AdhIFNβ基因组细胞凋亡率[(28.27±4.21)%]高于转染空载体组[(2.08±0.89)%]和对照组[(1.53±0.70)%](F=377.625,P<0.001),后2组细胞凋亡率比较差异无统计学意义。结论:腺病毒介导的hIFNβ基因能诱导体外肝癌细胞发生凋亡。
Abstract:
To study the effects of adenovirusmediated interferon(AdhIFNβ) gene on apoptosis of SMMC7721 cells in vitro.Methods:The HEK293A cells were infected with recombinant adenovirus AdhIFNβ and proceed with a largescale amplification of viral recombinant particles in HEK293A cells. Subsequently the virus was purification and concentrated. The titer of AdhIFNβ was determined by plaque assay in HEK293A cells. SMMC7721 cells were transduced by AdhIFNβ and AdGFP respectively,and the blank group was used as control. The hIFNβ expression was detected by using immunocytochemical method. The effects of recombinant AdhIFNβ gene inducing SMMC7721 cells apoptosis was detected by using Hochest 33342 fluorescent staining and flow cytometry.Results: AdhIFNβ was propagated in HEK293A cells and purified using AdenoXTM virus purification kits. The titers of AdhIFNβ reached 2×1011pfu/mL and more than 95% of SMMC7721 cells could be infected by 40 multiplicity of infection AdhIFNβ. The results of immunocytochemical method showed obvious hIFNβ protein in SMMC7721 cells after transfection. The apoptosis of SMMC7721 cells was obviously observed after transfected with recombinant AdhIFNβ by Hochest 33342 fluorescent staining and flow cytometry compared with that of the control cells, while the apoptosis of other groups cells did not observed obviously.The apoptosis rate of SMMC7721 cells transduced by AdhIFNβ[ (28.27±4.21)%]was higher than that of SMMC7721 cells transduced by AdGFP[ (2.08±0.89)%] and that of SMMC7721 cells [(1.53±0.70)%] (F=377.625,P<0.001), while the difference is no significance in statistics science between the apoptosis rate of the later groups.Conclusion: Adenovirusmediated interferonbeta gene could induce human hepatocellular carcinoma cells apoptosis in vitro.

参考文献/References:

[1]邓怀慈, 张晓智, 张龙. 反式维甲酸和干扰素γ协同作用对人胶质瘤细胞系SHG44增殖、凋亡及侵袭力的影响[J]. 西安交通大学学报:医学版,2004,25(4):401
[2]孟祥伟, 吴扬, 孙逊,等. 慢性丙型肝炎及肝硬化患者血清中干扰素α/β受体蛋白的测定[J].吉林大学学报:医学版,2005,31(6):946
[3]Johns TG, Mackay IR, Callister KA, et al. Antiproliferative potencies of interferons on melanoma cell lines and xenografts: higher efficacy of interferon beta[J]. J Natl Cancer Inst,1992, 84(15):1 185
[4]Shimizu M, Akiyama S, Ito K, et al. Effect on colon cancer cells of human interferonbeta gene entrapped in cationic multilamellar liposomes[J]. Biochem Mol Biol Int,1998,44(6):1 235
[5]Yoshida J, Mizuno M, Fujii M, et al. Human gene therapy for malignant gliomas (glioblastoma multiforme and anaplastic astrocytoma) by in vivo transduction with human βinterferon gene using cationic liposomes[J]. Hum Gene Ther,2004,15(1):77
[6]Kageshida T, Mizuno M, Ono T, et al. Growth inhibition of human malignant melanoma transfected with the human interferonβ gene by means of cationic liposomes[J]. Melanoma Res,2001,11(4):337
[7]Nakanishi H, Mizutani Y, Kawauchi A, et al.Significant antitumoral activity of cationic multilamellar liposomes containing human IFNβ gene against human renal cell carcinoma[J]. Clin Cancer Res,2003,9(3):1 129
[8]Nakahara N, Pollack IF,Storkus WJ, et al. Effective induction of antiglioma cytotoxic T cells by coadministration of interferonbeta gene vector and dendritic cells[J]. Cancer Gene Ther,2003,10(7):549
[9]Natsume A, Tsujimura K, Mizuno M, et al.IFNbeta gene therapy induces systemic antitumor immunity against malignant glioma[J].J Neurooncol,2000,47(2):117
[10]Natsume A, Mizuno M, Ryuke Y, et al.Antitumor effect and cellular immunity activation by murine interferonbeta gene transfer against intracerebral glioma in mouse[J].Gene Ther,1999,6(9):1 626
[11]Gamero AM, Larner AC. Vanadate facilitates interferon alphamediated apoptosis that is dependent on the Jak/Stat pathway[J]. J Biol Chem, 2001, 276(17):13 547
[12]Tomoda K, Ohishi N, Kikkawa F, et al. Cationic multilamellar liposomemediated human interferonbeta gene transfer into cervical cancer cell[J].Anticancer Res, 1998,18(3A):1 367
[13]Nakamoto N, Higuchi H, Kanamori H, et al. Cyclooxygenase2 inhibitor and interferonbeta synergistically induce apoptosis in human hepatoma cells in vitro and in vivo[J].Int J Oncol,2006,29(3):625
[14]Zitzmann K, Brand S, De Toni EN, et al. SOCS1 silencing enhances antitumor activity of type ⅠIFNs by regulating apoptosis in neuroendocrine tumor cells[J].Cancer Res, 2007,67(10):5 025
[15]Micali OC, Cheung HH, Plenchette S, et al. Silencing of the XAF1 gene by promoter hypermethylation in cancer cells and reactivation to TRAILsensitization by IFNbeta[J]. BMC Cancer,2007,7:52

相似文献/References:

[1]李振伟),李月白),王义生)#.靶向PPARγRNA干扰对激素诱导家兔骨髓基质细胞成脂分化的影响*[J].郑州大学学报(医学版),2009,(02):272.
 LI Zhenwei),LI Yuebai),WANG Yisheng).Effects of RNAi for target on PPARγ to suppress adipogenic differentiation of MSCs of rabbit induced by steroid[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2009,(06):272.
[2]张红飞),李军锋),户义),等.重组表达乙肝病毒x蛋白的腺病毒构建及在HepG2细胞的表达*[J].郑州大学学报(医学版),2012,(04):450.
 ZHANG Hongfei,LI Junfeng,HU Yi,et al.Construction and expression of recombinant HBxexpressed adenovirus and HBx expression in HepG2[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2012,(06):450.
[3]程剑△,沈晓洁,刘艳.携带人IL10基因的腺病毒载体的构建及其对大鼠肝细胞损伤的保护作用*[J].郑州大学学报(医学版),2013,(04):477.
 CHENG Jian,SHEN Xiaojie,LIU Yan.Construction of adenovirus vector carrying human interleukin10 gene and its protection effects on rat hepatocytes during liver injury[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2013,(06):477.

备注/Memo

备注/Memo:
*留学人员科技活动项目择优基金资助项目;河南省科技厅重点攻关基金资助项目0523034400 #通讯作者,男,1966年生,博士,副教授,研究方向:消化系统肿瘤,Email:cqguo6610@hotmail.com
更新日期/Last Update: 2010-05-14