论文部分内容阅读
目的：研究腺病毒／ＨＳＶ１－ｔｋ／ＧＣＶ、ＡＣＶ系统对卵巢上皮癌细胞旁观者效应。方法：常规重组腺病毒扩增纯化后，测病毒滴度。Ｘ－ｇａｌ染色法测定转染效率，ＰＣＲ检测ＨＳＶ１－ｔｋ的表达，混合细胞培养实验观察旁观者效应。结果：重组腺病毒扩增纯化后测定滴度分别为Ａｄ／ｔｋ*“６２×１０９ｐｆｕ／ｍｌ，ＡｄＬａｃＺ*”１４×１０９ｐｆｕ／ｍｌ。当感染复数为１００时，近１００％的细胞可被感染。ＰＣＲ结果显示，Ａｄｔｋ转染ＴＹＫ后，可见４０４ｂｐ的特异性条带，在同一ＭＯＩ中，有１０％的Ａｄｔｋ／ＴＹＫ细胞存在，ＧＣＶ就可杀死近５０％左右的混合细胞。ＡＣＶ的作用约为ＧＣＶ的１／１０。结论：重组腺病毒Ａｄ／ｔｋ可有效地感染人卵巢上皮癌细胞，腺病毒／ＨＳＶ１ｔｋ／ＧＣＶ、ＡＣＶ系统存在着旁观者效应。
Objective: To study the bystander effect of adenovirus / HSV1-tk / GCV and ACV system on epithelial ovarian cancer cells. Methods: After routine amplification of recombinant adenovirus, virus titer was measured. The transfection efficiency was determined by X-gal staining, the expression of HSV1-tk was detected by PCR, and the bystander effect was observed by mixed cell culture. Results: The titer of recombinant adenovirus was Ad / tk * "62 × 109pfu / ml, AdLacZ * 14 × 109pfu / ml, respectively. When the multiplicity of infection is 100, nearly 100% of the cells can be infected. PCR results showed that 404kb specific bands were found after TYK transfection with Adtk, and about 10% Adtk / TYK cells existed in the same MOI. GCV can kill about 50% mixed cells. The role of ACV is about 1/10 of GCV. Conclusion: Ad / tk recombinant adenovirus can effectively infect human ovarian epithelial carcinoma cells, adenovirus / HSV1-tk / GCV, ACV system exists bystander effect.