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Effect on proliferation and apoptosis of t24 cell lines via silencing DNMt1 with RNA interference

Frontiers of Medicine 2008年第4期2卷 374-379页

作者： Shilong ZHANG Fuqing ZENG Shibo PENG Liang WANG Department of Urology Union HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430022China

Expression of DNA methyltransferase 1(DNMt1),which plays an important role on aberrantly methylated CpG in the promoter regions of tumor sup-pressor genes(tSGs),is higher in bladder cancer cells than in normal bladder ***,its overexpression is closely related to tumor *** this study,the eukaryotic vector pshRNA-DNMt1 was constructed and transfected into t24 *** of DNMt1 mRNA and protein were detected by reverse transcription-polymerase chain reaction(Rt-PCR)and western *** to the blank control at the 24th,48th and 72nd hour after transfection of pshRNA-DNMt1,the inhibitory rates of DNMt1 mRNA levels in t24 cells were 28.44%,52.48%,70.91%,*** of DNMt1 proteins were 24.27%,57.79%,and 77.74%,*** and apoptosis were assayed by Mtt and flow cytometry with Annexin-V-FItC/PI *** growth inhibition rates of pshRNA-DNMt1 at the 24th,48th and 72nd hour after transfection of pshRNA-DNMt1 were(4.34¡0.76)%,(9.87¡1.54)%and(13.78¡1.93)%,*** were statistically significant differ-ences between pshRNA-DNMt1 and the control blank at each time points(P,0.01);24,48 and 72 hours after t24 cells were transfected by pshRNA-DNMt1,the apoptosis rates of pshRNA-DNMt1 were(3.87¡0.81)%,(8.69¡1.23)%and(11.46¡1.24)%,respectively(P,0.01 vs blank control).Based on this case,our conclu-sion is that the recombinant plasmid pshRNA-DNMt1 can silence the expression of gene DNMt1 mRNA and protein effectively,and to some extent,it also can inhibit the proliferation of bladder cancer cell and promote the cellular apoptosis.

关键词： DNA methyltransferase 1 RNA interference apoptosis t24 cell line proliferation

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Apoptosis of bladder transitional cell carcinoma t24 cells induced by adenovirus-mediated inducible nitric oxide synthase gene transfection

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Chinese Journal of Cancer Research 2013年第5期25卷 593-599页

作者： Jing tan Qing Zeng Xian-Zheng Jiang Le-Ye He Jin-Rong Wang Kun Yao Chang-Hui Wang Department of Urology The Third Xiangya Hospital of Central South University Department of Urology The Third Xiangya HospitalYue-lu District

Objectives：to investigate the effects of adenovirus-mediated inducible nitric oxide synthase gene transfection on bladder transitional cell carcinoma t24 cells,and to provide novel insights and approaches to clinical therapies against bladder transitional cell ***：Firstly,construct recombinant adenovirus vector pAd-iNOS of iNOS,followed by transfection of pAd-iNOS into HECK293 packaging ***,harvest recombinant adenovirus rAd-iNOS after amplification and purification ***,transfect the recombinant adenovirus rAd-iNOS into human bladder carcinoma t24 cells and examine the effect of rAd-iNOS transfection on apoptosis of t24 and possible ***：As shown by this study,the recombinant adenovirus rAd-iNOS was constructed *** virus titer was 5.8×108 PFU/mL and recombinant was verified by PCR *** of adenovirus rAd-iNOS into t24 cells could induce secretion of high NO concentration,P53 protein expression upregulation,as well as promotion oft24 cell ***：the transfection of human bladder carcinoma t24 cells from recombinant adenovirus rAdiNOS was confirmed to induce intracellular iNOS over-expression,high production of NO,up-regulation of intracellular P53 expression and promotion of cell apoptosis.

关键词： Bladder cancer t24 cell line iNOS gene gene therapy

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