恢复connexin26表达联合载酵母菌胞嘧啶脱氨酶自杀基因纳泡杀灭膀胱癌细胞

陈思乡; 姜庆; 张俊勇

doi:10.16016/j.1000-5404.201710163

您当前的位置：

首页 >

文章列表页 >

恢复connexin26表达联合载酵母菌胞嘧啶脱氨酶自杀基因纳泡杀灭膀胱癌细胞

基础医学 | 更新时间：2026-03-02

- 恢复connexin26表达联合载酵母菌胞嘧啶脱氨酶自杀基因纳泡杀灭膀胱癌细胞
- Efficacy of cationic nanomicrobubbles carrying connexin 26 gene and yeast cytosine deaminase/5-fluorocytosine suicide gene system combined with ultrasonic irradiation in killing bladder cancer cells in vitro
- 第三军医大学学报 2018年40卷第4期页码：322-327
- 作者机构：
  
  400016 重庆, 重庆医科大学附属第二医院泌尿外科Department of Urologic Surgery, the Second Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China
- 作者简介：
- 基金信息：
  
  重庆市卫生计生委重点科研项目(2013-1-020)
- DOI：10.16016/j.1000-5404.201710163
  中图分类号： R394-33;R394.3;R737.14
- 收稿：2017-10-24，
  
  纸质出版：2018-02-28
- 稿件说明：
移动端阅览
陈思乡, 姜庆, 张俊勇. 恢复connexin26表达联合载酵母菌胞嘧啶脱氨酶自杀基因纳泡杀灭膀胱癌细胞[J]. 第三军医大学学报, 2018,40(4):322-327.

Sixiang CHEN, Qing JIANG, Junyong ZHANG. Efficacy of cationic nanomicrobubbles carrying connexin 26 gene and yeast cytosine deaminase/5-fluorocytosine suicide gene system combined with ultrasonic irradiation in killing bladder cancer cells in vitro[J]. Journal of Third Military Medical University, 2018, 40(4): 322-327.
陈思乡, 姜庆, 张俊勇. 恢复connexin26表达联合载酵母菌胞嘧啶脱氨酶自杀基因纳泡杀灭膀胱癌细胞[J]. 第三军医大学学报, 2018,40(4):322-327. DOI： 10.16016/j.1000-5404.201710163.

Sixiang CHEN, Qing JIANG, Junyong ZHANG. Efficacy of cationic nanomicrobubbles carrying connexin 26 gene and yeast cytosine deaminase/5-fluorocytosine suicide gene system combined with ultrasonic irradiation in killing bladder cancer cells in vitro[J]. Journal of Third Military Medical University, 2018, 40(4): 322-327. DOI： 10.16016/j.1000-5404.201710163.

摘要

目的

分析载双基因的阳离子纳泡联合超声靶向微泡破坏技术进行基因转染的有效性，探索恢复或上调膀胱癌细胞的缝隙连接蛋白connexin26(Cx26)表达，能否增强自杀基因系统(yeast cytosine deaminase /5-fluorocytosine，YCD/5-FC)的旁观者效应，提高杀灭肿瘤细胞的效率。

方法

阳离子纳泡结合超声辐照(US)转染人膀胱癌T24细胞，荧光显微镜及流式细胞仪观测转染效率；qRT-PCR和Western blot检测质粒转染后的mRNA或蛋白相对表达量。将实验分为无处理空白对照、载pcDNA3.1-EGFP纳泡组、载Cx26纳泡组、载YCD纳泡组、载YCD+Cx26纳泡组；通过流式细胞术观察恢复Cx26表达后对膀胱癌细胞凋亡的影响。

结果

qRT-PCR、Western blot显示纳泡结合超声辐照成功将目的基因转染并有效表达。恢复Cx26表达后，载YCD+Cx26纳泡组的细胞凋亡率为(60.68±2.61)%，明显高于单载YCD纳泡组的(46.42±2.13)%，差异有统计学意义(

0.01)。

结论

恢复缝隙连接蛋白Cx26表达，可改善细胞间通讯连接，加强自杀基因系统YCD/5-FC的旁观者效应，促进肿瘤细胞凋亡，提高杀灭膀胱癌细胞的效率。

Abstract

Objective

To test the efficacy of cationic nanomicrobubbles carrying connexin 26 gene and yeast cytosine deaminase/5-fluorocytosine (YCD/5-FC) suicide gene system combined with ultrasonic irradiation in enhancing the cell killing effect in bladder cancer cells.

Methods

Human bladder cancer T24 cells were exposed to cationic nanomicrobubbles carrying both connexin 26 plasmid and the YCD/5-FC suicide gene system followed by ultrasonic irradiation. The gene transduction efficiency was assessed using fluorescence microscopy and flow cytometry

and qRT-PCR and Western blotting were used to verify the expression of the target genes. The effect of nanomicrobubble-mediated transduction of YCD/5-FC and connexin 26 genes on apoptosis of bladder cancer cells was detected by flow cytometry.

Results

qRT-PCR and Western blotting showed that the target genes were successfully transduced and efficiently expressed in the cells. The cells with nanomicrobubble-mediated transduction of both connexin 26 and YCD/5-FC genes showed an apoptotic rate of (60.68±2.61)%

which was significantly higher than the rate of (46.42±2.13)% in the cells transduced with YCD gene alone (

0.01).

Conclusion

Restoration of connexin 26 expression in bladder cancer cells can improve the intercellular communication

enhance the bystander effect of YCD/5-FC suicide gene system

promote cell apoptosis

and thus markedly improve the efficiency of cell killing.

关键词

Keywords

references

ANTONI S, FERLAY J, SOERJOMATARAM I, et al . Bladder cancer incidence and mortality: A global overview and recent trends[J ] . Eur Urol, 2017, 71(1): 96-108. DOI: 10.1016/j.eururo.2016.06.010.

MIYAZAKI J, NISHIYAMA H, Epidemiology of urothelial carcinoma[J]. Int J Urol, 2017, 24(10): 730-734.DOI: 10.1111/iju.13376.

NAVARRO S A, CARRILLO E, GRINAN-LISON C, et al . Cancer suicide gene therapy: a patent review[J ] . Expert Opin Ther Pat, 2016, 26(9): 1095-1104. DOI: 10.1080/13543776.2016.1211640.

MA S L, YANG X P, LIU L, et al . Human sulfatase-1 improves the effectiveness of cytosine deaminase suicide gene therapy with 5-fluorocytosine treatment on hepatocellular carcinoma cell line hepG2 in vitro and in vivo[J ] . Chin Med J, 2015, 128(10): 1384. DOI: 10.4103/0366-6999.156800.

YATA V K, GOPINATH P, GHOSH S S. Emerging implications of nonmammalian cytosine deaminases on cancer therapeutics[J]. Appl Biochem Biotech, 2012, 167(7): 2103-2116. DOI: 10.1007/s12010-012-9746-0.

ZHANG J, KALE V, CHEN M.Gene-directed enzyme prodrug therapy[J]. AAPS J, 2015, 17(1): 102-110. DOI: 10.1208/s12248-014-9675-7.

ZHU Y, ZONG L, MEI L, et al . Connexin26 gap junction mediates miRNA intercellular genetic communication in the cochlea and is required for inner ear development[J ] . Sci Rep, 2015, 5: 15647. DOI: 10.1038/srep15647.

DE TOLEDO S M, BUONANNO M, HARRIS A L, et al . Genomic instability induced in distant progeny of bystander cells depends on the connexins expressed in the irradiated cells[J ] . Int J Radiat Biol, 2017, 93(10): 1182-1194. DOI: 10.1080/09553002.2017.1334980.

LI X, SU Y, PAN J, et al . Connexin 26 is down-regulated by KDM5B in the progression of bladder cancer[J ] . Int J Mol Sci, 2013, 14(4): 7866-7879. DOI: 10.3390/ijms14047866.

WANG L, ZHANG M, TAN K, et al . Preparation of n anobubbles carrying androgen receptor siRNA and their inhibitory effects on androgen-independent prostate cancer when combined with ultrasonic irradiation[J ] . PLoS ONE, 2014, 9(5): e96586. DOI: 10.1371/journal.pone.0096586.

CAO S, ZHOU Q, CHEN J L, et al . Enhanced effect of nuclear localization signal peptide during ultrasound-targetedmicrobubble destruction-mediated gene transfection[J ] . Mol Med Rep, 2017, 16(1): 565-572. DOI: 10.3892/mmr.2017.6661.

ZHOU Y, GU H, XU Y, et al . Targeted antiangiogenesis gene therapy using targeted cationic microbubbles conjugated with CD105 antibody compared with untargeted cationic and neutral microbubbles[J ] . Theranostics, 2015, 5(4): 399-417. DOI: 10.7150/thno.10351.

WOOD A K, SEHGAL C M.A review of low-intensity ultrasound for cancer therapy[J]. Ultrasound Med Biol, 2015, 41(4): 905-928.DOI: 10.1016/j.ultrasmedbio.2014.11.019.

SMELSER W W, AUSTENFELD M A, HOLZBEIERLEIN J M, et al . Where are we with bladder preservation for muscle-invasive bladder cancer in 2017[J ] . Indian J Urol, 2017, 33(2): 111-117. DOI: 10.4103/iju.IJU_279_16.

阳孟君, 王洛夫, 郭燕丽, 等, 携载G250单克隆抗体的纳米微泡靶向于肾细胞癌的体内外实验研究[J].第三军医大学学报, 2016, 38(2): 112-118. DOI: 10.16016/j.1000-5404.201508106.

YANG M J, WANG L F, GUO Y L, et al . In vivo and in vitro study of nanoscale microbubbles with G250 monoclonal antibodies targeting to renal cell carcinoma[J ] . J Third Mil Med Univ, 2016, 38(2): 112-118. DOI: 10.16016/j.1000-5404.201508106.

ZHANG W, SHOU W D, XU Y J, et al .Low-frequency ultrasound-induced VEGF suppression and synergy with dendritic cell-mediated anti-tumor immunity in murine prostate cancer cells in vitro[J ] . Sci Rep, 2017, 7(1): 5778. DOI: 10.1038/s41598-017-06242-8.

LI C, YANG Z, DU Y, et al . BCMab1, a monoclonal antibody against aberrantly glycosylated integrin α3β1, has potent antitumor activity of bladder cancer in vivo[J ] . Clin Cancer Res, 2014, 20(15): 4001-4013.DOI: 10.1158/1078-0432.CCR-13-3397.

WU K, YANG L, HUANG Z, et al . A double suicide gene system driven by vascular endothelial growth factor promoter selectively kills human hepatocellular carcinoma cells[J ] . Oncol Lett, 2016, 11(5): 3152-3160. DOI: 10.3892/ol.2016.4360.

XIAO J, ZHANG G, QIU P, et al . Tanshinone ⅡA increases the bystander effect of herpes simplex virus thymidine kinase/ganciclovir gene therapy via enhanced gap junctional intercellular communication. PLoS ONE, 2013. 8(7): e67662. DOI: 10.1371/journal.pone.0067662.

XIAO J, ZHANG G, LI B, et al . Dioscin augments HSV-tk-mediated suicide gene therapy for melanoma by promoting connexin-based intercellular communication[J ] . Onco-target, 2017, 8(1): 798-807. DOI: 10.18632/oncotarget.13655.

SUN P, LIU Y, YING H, et al .Action of db-cAMP on the bystander effect and chemosensitivity through connexin 43 and Bcl-2-mediated pathways in medulloblastoma cells[J ] . Oncol Rep, 2012, 28(3): 969-976. DOI: 10.3892/or.2012.1900.

TAKAHASHI M, VALDES G, HIRAOKA K, et al . Radiosensitization of gliomas by intracellular generation of 5-fluorouracil potentiates prodrug activator gene therapy with a retroviral replicating vector[J ] . Cancer Gene Ther, 2014, 21(10): 405-410. DOI: 10.1038/cgt.2014.38.

浏览量

下载量

CSCD

文章被引用时，请邮件提醒。

提交

工具集

关联资源

暂无数据