抗HPV16E6核酶对宫颈癌细胞免疫学特性影响的研究

目的 研究特异性抗体HPV16E6核酶的转染对宫颈癌细胞免疫学特性的影响。方法 以脂质体法将抗HPV16E6-ribozyme、空载体质粒分别导入CaSKi细胞，命名为CaSKi、CaSKi-P细胞。流式细胞仪分别检测3种细胞HLA－1、HL2－2、B7－1和B7－2基因的表达，分析CaSKi细胞转染酶后免疫学特性的变化。诱导制备NK、LAK、CD3AK和肿瘤细胞共激活杀伤细胞，检测各种免疫细胞对CaSKi-R、CaSKi-P、CaSKi细胞的杀伤效应。结果 CaSKi和CaSKi-p细胞中HLA－2、B7－1、B7－2表达都很低，两者差异无显著性。CaSKi-R中HLA－2、B7－1、B7－2表达率均明显升高。3种细胞中HLA－1表达率都很高。NK、LAK、CD3AK细胞对CaSKi-R细胞伤率显著高于CaSKi细胞，CaSKi、CaSKi-R分别与rIL－2共激活杀伤细胞（称CASKI和CASKI－R）的样伤活性无明显差别，对CaSKi-R的杀伤活性高于CaSKi细胞。而各种免疫细胞对CaSKi-P细胞的杀伤率与CaSKi细胞差异无显著性。结论 抗HPV16E6－nrbozyme的导入能使宫颈癌CaSKi细胞易于被机体免疫系统识别杀伤，难于免疫逃避，但不能增强其免疫原性。

The effect of anti-HPV16E6-ribozyme on immune phenotypes of cervical carcinoma cell line

Objective To study the immune characterization of the cultured cervical cancer cell line transfected with anti HPV16E6 ribozyme, and to explore the possibility of using ribozyme in immune therapy and gene therapy of cervical cancer. Methods With the method of lipofectin transfection, the anti HPV16E6 ribozyme and empty eucaryotic expressing plasmids were transfected into CaSKi cell, which are then named as CaSKi R, CaSKi P respectively. The expression of ribozyme in transfected cells was observed. The expression of HLA 1, HLA 2, B7 1, B7 2 genes was studied by flow cytometry. NK, LAK, K lymphocyte stimulated by tumor cells and rIL 2, and CD3AK cells were induced, and their cytotoxicity were detected in CaSKi R, CaSKi P and CaSKi cell. Results Anti HPV16E6 ribozyme could be stably expressed in transfected CaSKi cells. The result of flow cytometric analysis showed that anti HPV16E6 ribozyme could improve the expression of HLA 2, B7 1, B7 2 genes on CaSKi R cells, but not in the CaSKi P cell. HLA 1 was highly expressed in all three kinds of cells. Compared with CaSKi cells, the sensitivity of CaSKi R to NK cells was enhanced. The cytotoxicity of LAK and CD3AK cells to CaSKi R was also much higher than that to CaSKi P and CaSKi cells. The cytotoxicity of killer cells, which were coactived by CaSKi and CaSKi R cells with rIL 2 respectively (CASKI and CASKI R), were similar. The killing rate of CASKI and CASKI R to CaSKi R cells was obviously higher than to CaSKi cells. Conclusion Anti HPV16E6 rivozyme can enhance the expression of HLA 2, B7 1, B7 2 genes, and increase the sensitivity of CaSKi cells to immune cells, and so CaSKi cells would hardly escape from host immune surveillance. However, anti HPV16E6 ricozyme fails to enhance the immunogenicity of CaSKi cells.