【摘要】 目的　探讨共刺激分子B7基因能否在体内诱导小鼠抗宫颈癌主动免疫应答。方法　将B7基因转染小鼠宫颈癌细胞株即U14,建立高表达B7的U14细胞株即B7+U14。随后分 4组进行体内实验 :(1)实验A组 :将B7+U14(1× 10 7个细胞 )接种于同系 6 15小鼠的右侧背部皮下 (n =6 )。(2 )实验B组 :用B7+U14(1× 10 6个细胞 )皮下注射免疫 6 15小鼠 ,7d后将U14(1× 10 7个细胞 )接种于免疫后的小鼠背部皮下 (n =6 )。 (3)对照A组 :在 6 15小鼠右侧背部皮下接种U14(1× 10 7个细胞 ) ,其余条件同实验A组。 (4 )对照B组 :用U14(1× 10 6个细胞 )免疫 6 15小鼠 ,其余条件同实验B组。观察 4组小鼠的成瘤情况、生存时间 ;体外分别检测经B7+U14和U14免疫后的小鼠 (n =4× 2 )T淋巴细胞对肿瘤细胞的杀伤作用。结果　 (1)小鼠皮下接种B7+U14后 ,体内的致瘤能力明显降低 (P<0 0 1)。(2 )用B7+U14免疫后再皮下接种U14可有效预防移植瘤产生 (P <0 0 1)。(3)体外检测经B7+U14和U14免疫后的小鼠T淋巴细胞杀伤肿瘤细胞作用 ,前者明显高于后者 (P <0 0 1)。结论　B7基因转染小鼠宫颈癌细胞株U14能诱导机体有效的抗肿瘤主动免疫应答 ,提示应用B7基因转染法可能成为临床治疗宫颈癌的有效方法更多还原

【Abstract】 Objective To investigate the effects of B 7 costimulatory molecule on inducing anti tumor immune response to cervical carcinoma in vivo Methods We transfected mouse B 7 gene into murine cervix carcinoma cell line U14 by electroporation, and obtained several high expressed mB 7 U14 cell clonal strains (B 7 +U14) detected by reverse transcription polymerase chain reaction (RT PCR) In vivo experiments: (1) 1×10 7 B 7 +U14 cells were inoculated into the back of inbred 615 strain mice by subcutaneously injection to determine their tumorigenicity ( n =6) (2) The mice primed by B 7 +U14 (1×10 6) cells were re challenged with 1×10 7 wild type U14 to observe the immune protection of these mice against the wild type U14 ( n =6) As control, (3) wild type U14 cells were inoculated the same as the experimental group (1) ( n =6) (4) The mice both primed and re challenged with 1×10 7 wild type U14 the same as the experimental group (2) ( n =6) All mice lifetime and tumor sizes were recorded In vitro cytotoxiaty assay: the mice were immunized with B 7 +U14 or the wild type U14 by intraperitoneal injection ( n =4×2) and two weeks late those mice spleen cells were obtained and cultured for two days The cytotoxiaty of these cells against the wild U14 was detected by methyl thiazolyl tetrazolium assay Results RT PCR showed positive results in B 7 +U14 cells, while negative in U14 cells In vivo experiment: (1) after the inoculation of the B 7 +U14 cells into the back of inbred mice, they lost their tumorigenicity greatly compared to wild type U14 ( P <0 01) (2) Primed by B 7 +U14, mice protected themselves effectively against re challenged ( P <0 01). In vitro cytotoxiaty assay, the cytotoxic T lymphocytes (CTLs) induced by B 7 +U14 had a higher cytotoxiaty against the wild type U14 than that induced by wild type U14 ( P< 0 01) Conclusions Cervical carcinoma cells transfected with constimulatory molecules B 7 gene can decrease their tumorigenicity greatly, and induce anti tumor immune protection of inbred mice against wild type U14 cells re challenged The results suggest that to transfect B 7 gene into cervical cancer expression may be an effect method for treating cervical cancer in clinical.更多还原