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自杀基因系统HSV-TK/GCV联合声动力疗法对人肺癌细胞NCI-446的体外杀伤实验

李洋, 马程远, 里程楠, 钱东华

李洋, 马程远, 里程楠, 钱东华. 自杀基因系统HSV-TK/GCV联合声动力疗法对人肺癌细胞NCI-446的体外杀伤实验[J]. 肿瘤防治研究, 2013, 40(03): 226-231. DOI: 10.3971/j.issn.1000-8578.2013.03.002
引用本文: 李洋, 马程远, 里程楠, 钱东华. 自杀基因系统HSV-TK/GCV联合声动力疗法对人肺癌细胞NCI-446的体外杀伤实验[J]. 肿瘤防治研究, 2013, 40(03): 226-231. DOI: 10.3971/j.issn.1000-8578.2013.03.002
Li Yang, Ma Chengyuan, Li Chengnan, Qian Donghua. Effects of Treatment with Suicide Gene System HSV-TK/GCV Combined with Sonodynamic Therapy on Human Lung Cancer Cell NCI-446 in vitro[J]. Cancer Research on Prevention and Treatment, 2013, 40(03): 226-231. DOI: 10.3971/j.issn.1000-8578.2013.03.002
Citation: Li Yang, Ma Chengyuan, Li Chengnan, Qian Donghua. Effects of Treatment with Suicide Gene System HSV-TK/GCV Combined with Sonodynamic Therapy on Human Lung Cancer Cell NCI-446 in vitro[J]. Cancer Research on Prevention and Treatment, 2013, 40(03): 226-231. DOI: 10.3971/j.issn.1000-8578.2013.03.002

自杀基因系统HSV-TK/GCV联合声动力疗法对人肺癌细胞NCI-446的体外杀伤实验

详细信息
    作者简介:

    李洋(1979-),女,博士,主治医师,主要从事肺癌的分子生物学研究

    通讯作者:

    钱东华,E-mail:qiandonghua6016@163.com

  • 中图分类号: R734.2;R730.58

Effects of Treatment with Suicide Gene System HSV-TK/GCV Combined with Sonodynamic Therapy on Human Lung Cancer Cell NCI-446 in vitro

  • 摘要: 目的 探讨自杀基因系统HSV-TK/GCV联合声动力疗法对人小细胞肺癌NCI-446细胞的体外杀伤作用。方法利用荧光和RT-PCR检测方法,筛选出含HSV-TK基因的阳性克隆株NCI-446/TK。用MTT法检测不同浓度的GCV对NCI-446/TK细胞的杀伤效应,以及不同浓度混合NCI-446和NCI-446/TK细胞后的旁观者效应。同时,将HSV-TK/GCV自杀基因系统与声动力疗法进行联合,通过MTT法测定细胞存活率,并以流式细胞术检测细胞周期及凋亡率。结果当GCV浓度为0.2 mg/L时,筛选出的阳性克隆株NCI-446/TK细胞的生长受到明显的抑制(P<0.01),并且随着GCV浓度的递增,抑制效应越明显。当NCI-446/TK细胞占总细胞的10%时,有46%的混合培养细胞生长受到抑制(P<0.01),其抑制效应随NCI-446/TK细胞所占比例的增加而增加。将HSV-TK/GCV自杀基因系统与声动力疗法进行联合后,其细胞存活率明显低于单一治疗组(P<0.01),并且G0/G1期细胞所占比例及凋亡率均高于单一治疗组。结论HSV-TK/GCV自杀基因系统的杀伤效应随着GCV浓度的递增而增强,并且有明显的旁观者效应。与声动力疗法联合后,其作用效果优于单一治疗组。

     

    Abstract: Objective To observe the effects of suicide gene system HSV-TK/GCV combined with sonodynamic therapy on human lung cancer cell NCI-446 in vitro. Methods NCI-446/TK containing HSV-TK gene has been screened by the method of fluorescence and RT-PCR.MTT detected the damage effects of cells after NCI-446/TK treated with different concentrations of GCV.And bystander effect was also measured through mixed different concentrations of NCI-446 and NCI-446/TK. Meanwhile,combining HSV-TK/GCV suicide gene system and sonodynomic theragy,cell survival rate was tested by MTT,in addition,cell cycle and apoptotic rate were tested by Flow cytometry. Results With 0.2 mg/L of GCV,the growth of NCI-446/TK was inhibited significantly (P<0.01).With increased GCV,the inhibitory effect became obviously.When 10% NCI-446/TK in mixed cells,there was 46% total cells were inhibited.And also the inhibitory effect increased with increasing proportion of NCI-446/TK.After therapeutic alliance with suicide gene system HSV-TK/GCV and SDT,cell survival ratio was significantly lower than the groups of single treatment and the proportion of cells in G0/G1stage,as well as apoptotic rate,were also higher than the groups of single treatment. Conclusion The lethal effect of HSV-TK/GCV suicide gene system was enhanced with increase of GCV concentration and bystander effect was also significant.When combined with SDT,the effect was better than a single treatment group.

     

  • [1] Jemal A,Bray F,Center MM,et al.Global cancer statistics [J].CA Cancer J Clin,2011,61(2):69-90.
    [2] Ferlay J,Autier P,Boniol M,et al.Estimates of the cancer incidence and mortality in Europe in 2006 [J].Ann Oncol,2007,18(3):581-92.
    [3] Granville CA,Dennis PA.An overview of lung cancer genomics and proteomics [J].Am J Respir Cell Mol Biol,2005,32(3):169-76.
    [4] Mulshine JL,Sullivan DC.Clinical practice lung cancer screening [J].N Engl J Med,2005,352(26):2714-20.
    [5] Liu J.The progress of lung cancer treatment [J].Yi Xue Li Lun Yu Shi Jian,2011,24(1):25-7.[刘璟.肺癌治疗的进展[J].医学理论与实践,2011,24(1):25-7.]
    [6] The national cancer research office,national cancer registration center,ministry of health,disease prevention and control bureau.Cancer incidence and death in some city and county of Chinese[M].Volume 3 (1998-2002).Beijing:People's health publishing house,2007:26-164.[全国肿瘤防治研究办公室,全国肿瘤登记中心,卫生部疾病预防控制局.中国部分市、县恶性肿瘤的发病与死亡[M].第3卷(1998-2002).北京:人民卫生出版社,2007:26-164.]
    [7] Wang Q,He XR,Tian JH,et al.A meta analysis of aidi injection plus taxotere and cisplatin in the treatment of non-small cell lung cancer [J].Zhongguo Fei Ai Za Zhi,2010,13(11):1027-34.[王权,何曦冉,田金徽,等.艾迪联合紫杉醇和顺铂治疗晚期非小细胞肺癌的meta分析[J].中国肺癌杂志,2010,13(11):1027-34.]
    [8] Hasani A,Leighl N.Classification and toxicities of vascular disrupting agents [J].Clin Lung Cancer,2011,12(1):18-25.
    [9] Curiel DT,Gerritsen WR,Krul MR.Progress in cancer gene therapy [J].Cancer Gene Ther,2000,7(8):1197-9.
    [10] Tanaka M,Inase N,Mlyake S,et al.Neurons pecific enolasepromoter for suicidegene therapy in small lung cancinoma [J].Anticancer Res,2001,21(1A):291-4.
    [11] Rosenthal I,Sostaric JZ,Riesz P.Sonodynamic therapy-a review of the synergistic effects of drugs and ultrasound [J].Ultrason Sonochem,2004,11(6):349-63.
    [12] Li LZ,Li J.Research progress of sonodymanic therapy on tumour [J].Yi Liao Wei Sheng Zhuang Bei,2008,29(2):37-8.[李陆振,黎静.声动力治疗肿瘤的研究进展[J].医疗卫生装备,2008,29(2):37-8.]
    [13] Qian DH,Ma ZS,Sun XF,et al.Effects of ultrasound combined with hematoporphyrin on NCI-446,PLA-80IC,PLA-801D of human lung cancer cell lines in vitro [J].Jilin Da Xue Xue Bao(Yi Xue Ban),2007,204(2):149-51.[钱东华,马忠森,孙晓峰,等.超声激活血卟啉对体外培养的人肺癌细胞NCI-446、PLA-801C和PLA-801D 的杀伤作用 [J].吉林大学学报(医学版),2007,204(2):149-51.]
    [14] Liang YC,Cheng L,Ye QN.Research development of cancer gene therapy [J].Sheng Wu Ji Shu Tong Xun,2012,23(3):436-9.[梁迎春,程龙,叶棋浓.肿瘤基因治疗的研究进展 [J].生物技术通讯,2012,23(3):436-9.]
    [15] Mori K,Iwata J,Miyazaki M,et al.Bystander killing effect of tymidine kinase gene-transduced adult bone marrow stromal cells with ganciclovir on malignant glioma cells [J].Neurol Med Chir (Tokyo),2010,50(7):545-53.
    [16] Wang Y,Canine BF,Hatefi A.HSV-TK/GCV cancer suicide gene therapy by a designed recombinant multifunctional vector [J].Nanomedicine,2011,7(2):193-200.
    [17] Vlachaki MT,Chhikara M,Aguilar L,et al.Enhanced therapeutic effect of multiple injections of HSV-TK + GCV gene therapy in combination with ionizing radiation in a mouse mammary tumor model [J].Int J Radiat Oncol Biol Phys,2001,51(4):1008-17.
    [18] Van Dillen IJ,Mulder NH,Sluiter WJ,et al.Consequences of chemoresistance for the herpes simplex virus thymidine kinase/ganciclovir-induced bystander effect in a human small cell lung cancer cell line model [J].Anticancer Res,2005,25(1A):255-61.
    [19] Feng R,Wang ZB.Practical ultrasonic therapy [M].Beijing:Scientific and Technical Documentation Press,2002:23-5.[冯若,王志彪.实用超声治疗学[M].北京:科学技术文献出版社,2002:23-5.]
    [20] Miyoshi N,Igarashi T,Riesz P.Evidence against singlet oxygen formation by sonolysis of aqueous oxygen-saturated solutions of Hematoporphyrin and rose bengal.The mechanism of sonodynamic therapy [J].Ultrason Sonochem,2000,7(3):121-4.
    [21] Yumita N,Sakata I,Nakajima S,et al.Ultrasonically induced cell damage and active oxygen generation by 4-formyloximeetylidene-3-hydroxyl-2-vinyl-deuterio-porphynyl (IX)-6-7-diaspartic acid:on them echanism of sonodynamic activation [J].Biochim Biophys Acta,2003,1620(1-3):179-84.
    [22] Jeffers RJ,Feng RQ,Fowlkes JB,et al.Dimethylformamide as an enhancer of cavitation- induced cell lysis in vitro [J].J Acoust Soc Am,1995,97(1):669-76.
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出版历程
  • 收稿日期:  2012-05-14
  • 修回日期:  2012-09-10
  • 刊出日期:  2013-03-24

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