CAJ | 학술논문

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（1×107个细胞）接种于同系615小鼠的右侧背部皮下（n=6）。(2)实验B组：用B7+U14（1×106个细胞）皮下注射免疫615小鼠，7 d后将U14（1×107个细胞）接种于免疫后的小鼠背部皮下（n=6）。(3)对照A组：在615小鼠右侧背部皮下接种U14（1×107个细胞），其余条件同实验A组。(4)对照B组：用U14（1×106个细胞）免疫615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<0.01）。(2)用B7+U14免疫后再皮下接种U14可有效预防移植瘤产生（P<0.01）。(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<0.01）。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。
목적탐토공자격분자B7기인능부재체내유도소서항궁경암주동면역응답。방법장B7기인전염소서궁경암세포주즉U14，건립고표체B7적U14세포주즉B7+U14。수후분4조진행체내실험：(1)실험A조：장B7+U14（1×107개세포）접충우동계615소서적우측배부피하（n=6）。(2)실험B조：용B7+U14（1×106개세포）피하주사면역615소서，7 d후장U14（1×107개세포）접충우면역후적소서배부피하（n=6）。(3)대조A조：재615소서우측배부피하접충U14（1×107개세포），기여조건동실험A조。(4)대조B조：용U14（1×106개세포）면역615소서，기여조건동실험B조。관찰4조소서적성류정황、생존시간；체외분별검측경B7+U14화U14면역후적소서（n=4×2）T림파세포대종류세포적살상작용。결과 (1)소서피하접충B7+U14후，체내적치류능력명현강저（P<0.01）。(2)용B7+U14면역후재피하접충U14가유효예방이식류산생（P<0.01）。(3)체외검측경B7+U14화U14면역후적소서T림파세포살상종류세포작용，전자명현고우후자（P<0.01）。결론 B7기인전염소서궁경암세포주U14능유도궤체유효적항종류주동면역응답，제시응용B7기인전염법가능성위림상치료궁경암적유효방법。
Objective　To investigate the effects of B7 costimulatory molecule on inducing anti-tumor immune response to cervical carcinoma in vivo. Methods　We transfected mouse B7 gene into murine cervix carcinoma cell line U14 by electroporation, and obtained several high-expressed mB7 U14 cell clonal strains (B7+U14) detected by reverse transcription polymerase chain reaction (RT-PCR). In vivo experiments: (1) 1×107 B7+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 B7+U14 (1×106) cells were re-challenged with 1×107 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 1×107 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 B7+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 B7+U14 cells, while negative in U14 cells. In vivo experiment: (1) after the inoculation of the B7+U14 cells into the back of inbred mice, they lost their tumorigenicity greatly compared to wild type U14 (P<0.01). (2) Primed by B7+U14, mice protected themselves effectively against re-challenged (P<0.01). In vitro cytotoxiaty assay, the cytotoxic T lymphocytes (CTLs) induced by B7+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 B7 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 B7 gene into cervical cancer expression may be an effect method for treating cervical cancer in clinical.