CAJ | 학술논문

目的观察Scopadalciol(SDC)在人端粒酶逆转录酶(hTERT)启动子调控腺病毒介导的单纯疱疹病毒胸腺激酶基因/丙氧鸟苷(HSV-tk/GCV)自杀基因系统对人膀胱癌细胞的体外杀伤作用中的增效作用.方法利用重组腺病毒Ad-hTERT-HSV-tk、GCV、SDC的不同组合作用于膀胱癌细胞253J和人成纤维细胞MRC-5,噻唑蓝(MTT)比色法观察细胞存活率;另外,利用携带Ad-hTERT-HSV-tk感染253J细胞和MRC-5细胞,加入不同组合和不同浓度的GCV和SDC,MTT法观察受染细胞的存活率.结果经重组腺病毒Ad-hTERT-HSV-tk感染的253J细胞,应用GCV处理后,253 J细胞能被特异性地杀伤,而MRC-5细胞不被特异性杀伤,253J细胞存活率为25.7%(P＜0.01),联合应用SDC(0.1 μmol)后,253J细胞的存活率又有明显降低,253J细胞存活率为2.3%(P＜0.01);不同剂量SDC对不同配伍浓度的Ad-hTERT-HSV·tk/GCV作用于膀胱肿瘤细胞253J后,随着剂量和浓度的增加,253J细胞的存活率呈降低趋势(P＜0.05),当GCV浓度为1 μmol/L,SDC剂量为0.04 μmol时,253J细胞的存活率为45.7%,当GCV浓度为100 μmol/L,SDC剂量为0.1 μmol时,253J细胞的存活率仅为2.3%,并有旁观者效应.结论在胸腺激酶依赖的GCV可以靶向杀伤人膀胱癌细胞的基础上,联合应用SDC后,对膀胱癌细胞的杀伤作用具有明显增效作用.
목적 관찰Scopadalciol(SDC)재인단립매역전록매(hTERT)계동자조공선병독개도적단순포진병독흉선격매기인/병양조감(HSV-tk/GCV)자살기인계통대인방광암세포적체외살상작용중적증효작용.방법 이용중조선병독Ad-hTERT-HSV-tk、GCV、SDC적불동조합작용우방광암세포253J화인성섬유세포MRC-5,새서람(MTT)비색법관찰세포존활솔;령외,이용휴대Ad-hTERT-HSV-tk감염253J세포화MRC-5세포,가입불동조합화불동농도적GCV화SDC,MTT법관찰수염세포적존활솔.결과 경중조선병독Ad-hTERT-HSV-tk감염적253J세포,응용GCV처리후,253 J세포능피특이성지살상,이MRC-5세포불피특이성살상,253J세포존활솔위25.7%(P＜0.01),연합응용SDC(0.1 μmol)후,253J세포적존활솔우유명현강저,253J세포존활솔위2.3%(P＜0.01);불동제량SDC대불동배오농도적Ad-hTERT-HSV·tk/GCV작용우방광종류세포253J후,수착제량화농도적증가,253J세포적존활솔정강저추세(P＜0.05),당GCV농도위1 μmol/L,SDC제량위0.04 μmol시,253J세포적존활솔위45.7%,당GCV농도위100 μmol/L,SDC제량위0.1 μmol시,253J세포적존활솔부위2.3%,병유방관자효응.결론 재흉선격매의뢰적GCV가이파향살상인방광암세포적기출상,연합응용SDC후,대방광암세포적살상작용구유명현증효작용.
Objective To investigate the enhanced effect of scopadulciol(SDC)on the killing effect on antitumor actions by Ad-hTERT-HSV-tk/GCV suicide gene system on bladder cancer cells in vitro.Methods Bladder cancer cell line 253J and human fibroblast cell line MRC-5 were transduced by Ad-hTERT-HSV-tk.followed by the therapy of the GCV in combination with SDC or the single GCV or the singh SDC.and the combination of GCV and SDC was constituted with different doses and different concentrations.The relative survival rate of cells in the presence of prodrug GCV and SDC was measured by using MTT method.Results GCV inhibited the growth of only 253J infected with Ad-hTERT-HSV-tk,but did not inhibit the growth ofMRC-5 significantly.The survival rate of 253J cells wag 25.7%(P＜0.01)with the difference being significant between the combination therapy and GCV monotherapy.After a combined use of GCV with a higher concentration of SDC(0.1 μmol/L),the survival rate of 253J cells was 2.3%(P＜0.01).,The survival of253J wag correlated with both the doses of SDC and the concentrationof GCV.Whell the dose of SDC was 0.04 ttmoI/L in combination witb 1 μmol/L GCV,the sumval rate of 253J was 45.7%:when the dose ofSDC was 0.1 μmol/L in combination with 100 μmol/L GCV,the surviral rate of 253J wss 2.3%(P＜0.05).The bystander effect was also significantly augmented by the combined treatment of GCV and SDC.Conclusion SDC was shown to be effective in the Ad-hTERT-HSV-tk/GCV administration system and improved the efficiency of the bystander effect of GCV in killing bladder cancer cells.