郑州大学学报(医学版)
鄭州大學學報(醫學版)
정주대학학보(의학판)
JOURNAL OF ZHENGZHOU UNIVERISTY(MEDICAL SCIENCES)
2010年
6期
1001-1004
,共4页
何炜%刘甲%郭玉琴%刘新郑
何煒%劉甲%郭玉琴%劉新鄭
하위%류갑%곽옥금%류신정
MAGE-1%树突状细胞%细胞毒性T淋巴细胞
MAGE-1%樹突狀細胞%細胞毒性T淋巴細胞
MAGE-1%수돌상세포%세포독성T림파세포
MAGE-1%dendritic cell%cytotoxicity T lymphocyte
目的：研究肿瘤相关抗原MAGE-1的抗肿瘤生物活性。方法：取健康人外周血,分离树突状细胞和T淋巴细胞。用脂质体介导法分别将质粒pcDNA3.1-MAGE-1和pcDNA3.1转染人肝癌SMMC-7721细胞株,设未转染的SMMC-7721细胞株为空白对照组,应用Real-timePCR检测3组细胞中MAGE-1 mRNA的表达;再分别用以上3种细胞的冻融抗原致敏树突状细胞,使其激活T淋巴细胞成为细胞毒性T淋巴细胞（CTL）。应用乳酸脱氢酶释放法检测CTL对野生型SMMC-7721细胞的杀伤活性。结果：pcDNA3.1-MAGE-1转染组MAGE-1 mRNA的表达量为（1.211±0.586）,空质粒转染组及未转染组分别为（0.412±0.021）和（0.452±0.317）,3组比较,差异有统计学意义（F=538.652,P〈0.001）。转染pcDNA3.1-MAGE-1重组质粒、pcDNA3.1空质粒的SMMC-7721细胞和未转染的野生型SMMC-7721细胞的冻融抗原所激活的CTL对野生型SMMC-7721细胞的杀伤活性分别为（48.26±2.47）%、（25.84±2.72）%和（26.27±0.81）%,3组比较,差异有统计学意义（F=139.607,P〈0.001）。结论：肿瘤相关抗原MAGE-1能够在体外诱导细胞抗肿瘤免疫反应。
目的：研究腫瘤相關抗原MAGE-1的抗腫瘤生物活性。方法：取健康人外週血,分離樹突狀細胞和T淋巴細胞。用脂質體介導法分彆將質粒pcDNA3.1-MAGE-1和pcDNA3.1轉染人肝癌SMMC-7721細胞株,設未轉染的SMMC-7721細胞株為空白對照組,應用Real-timePCR檢測3組細胞中MAGE-1 mRNA的錶達;再分彆用以上3種細胞的凍融抗原緻敏樹突狀細胞,使其激活T淋巴細胞成為細胞毒性T淋巴細胞（CTL）。應用乳痠脫氫酶釋放法檢測CTL對野生型SMMC-7721細胞的殺傷活性。結果：pcDNA3.1-MAGE-1轉染組MAGE-1 mRNA的錶達量為（1.211±0.586）,空質粒轉染組及未轉染組分彆為（0.412±0.021）和（0.452±0.317）,3組比較,差異有統計學意義（F=538.652,P〈0.001）。轉染pcDNA3.1-MAGE-1重組質粒、pcDNA3.1空質粒的SMMC-7721細胞和未轉染的野生型SMMC-7721細胞的凍融抗原所激活的CTL對野生型SMMC-7721細胞的殺傷活性分彆為（48.26±2.47）%、（25.84±2.72）%和（26.27±0.81）%,3組比較,差異有統計學意義（F=139.607,P〈0.001）。結論：腫瘤相關抗原MAGE-1能夠在體外誘導細胞抗腫瘤免疫反應。
목적：연구종류상관항원MAGE-1적항종류생물활성。방법：취건강인외주혈,분리수돌상세포화T림파세포。용지질체개도법분별장질립pcDNA3.1-MAGE-1화pcDNA3.1전염인간암SMMC-7721세포주,설미전염적SMMC-7721세포주위공백대조조,응용Real-timePCR검측3조세포중MAGE-1 mRNA적표체;재분별용이상3충세포적동융항원치민수돌상세포,사기격활T림파세포성위세포독성T림파세포（CTL）。응용유산탈경매석방법검측CTL대야생형SMMC-7721세포적살상활성。결과：pcDNA3.1-MAGE-1전염조MAGE-1 mRNA적표체량위（1.211±0.586）,공질립전염조급미전염조분별위（0.412±0.021）화（0.452±0.317）,3조비교,차이유통계학의의（F=538.652,P〈0.001）。전염pcDNA3.1-MAGE-1중조질립、pcDNA3.1공질립적SMMC-7721세포화미전염적야생형SMMC-7721세포적동융항원소격활적CTL대야생형SMMC-7721세포적살상활성분별위（48.26±2.47）%、（25.84±2.72）%화（26.27±0.81）%,3조비교,차이유통계학의의（F=139.607,P〈0.001）。결론：종류상관항원MAGE-1능구재체외유도세포항종류면역반응。
Aim：To observe the anti-tumor effects of tumor associated antigen MAGE-1 in vitro.Methods：DCs and T cells were separated from the peripheral blood of a healthy man. The SMMC-7721 cell line was transfected with the recombinant pcDNA3.1-MAGE-1 and the vector pcDNA3.1 with liposome and the expression level of MAGE-1 was checked by RT-PCR. DCs were divided into 3 groups, presenting the freeze-thaw antigen of SMMC-7721-pcDNA3.1-MAGE-1,SMMC-7721-pcDNA3.1 and wild type SMMC-7721 to naive T cells and induce them into CTLs. Taking wild type SMMC-7721 as target cells, the cytotoxicity of CTLs was detected by CytoTo 96 Non-Radioactive Cytotoxicity Assay.Results：The expression level of MAGE-1 in SMMC-7721 cells transfected with the recombinant pcDNA3.1-MAGE-1 was （1.211±0.586）, which of pcDNA3.1 transfected cells and wild type cells were （0.412±0.021） and （0.452±0.317） （F=538.652,P0.001）, respectively. The cytotoxicity of three groups of CTLs activated by freeze-thaw antigen of SMMC-7721-pcDNA3.1-MAGE-1,SMMC-7721-pcDNA3.1 and wild type SMMC-7721 was （48.26±2.47）%,（25.84±2.72）% and （26.27±0.81）%（F=139.607,P0.001）, respectively.Conclusion： The tumor associated antigen MAGE-1 is able to induce cellular immune responses in vitro.
