摘要
目的 观察反义寡核苷酸封闭鼠淋巴细胞H-2Kd基因后,对其表达的影响及淋巴因子激活杀伤细胞(LAK)的活性改变,探讨淋巴细胞Ⅰ类主要组织相容性抗原(MHC-Ⅰ)在肿瘤免疫中的作用。方法 将人工合成的H-2Kd起始区反义寡核苷酸作用于小鼠脾LAK,流式细胞仪检测作用前后H-2Kd的表达率。噻唑蓝法检测H-2Kd表达降低的LAK对肿瘤杀伤活性。结果 反义寡核苷酸(15μmoL/l)组H-2Kd蛋白的表达率由90.1％±4.4％下降至79.8％±2.5％(P<0.01),H-2Kd降低的LAK杀伤活性明显降低,对K562的杀伤活性由82.3％±3.1％降到60.2％±6.7％(P<0.01),对H22细胞株的杀伤活性由67.4％±2.8％降到53.2％±8.1％(P<0.01)。结论 反义寡核苷酸能够抑制小鼠LAK表面H-2Kd的表达,但不影响LAK增殖活性,因而可导致鼠LAK对同种及异种肿瘤细胞的杀伤活性降低。
Objective In order to understand the effect of MHC-I of host lymphocyte in tumor immune, observing proliferation and cytotoxicity of BALB/c spleen LAK cells which with silent the gene by H-2Kd antisense oligonucleotides ( ASON) . Methods ASON complementary to initiator of H-2Kd was applied to spleen LAK cells. The H-2Kd gene expression of treated and control cells were examined by Flow cytometry(FCM). The proliferation and cytotoxicity for H22 and K562 cell lines were evaluated in vitro with MTT colorimetric assay. Results ASON (15μmol/l) treatment for H-2Kd gene reduced targeted protein from 90. 1%±4.4% to 79. 8% ± 2. 5% (P < 0. 01). The cytotoxicity for K562 reduced from 82. 3% ± 3. 1% to 60. 2% ± 6. 7% ( P < 0. 01), and that for H22 cells reduced from 67.4% ± 2. 8% to 53. 2%± 8. 1% (P< 0. 01 ). Conclusion The reduced expression level of H-2Kd gene silenced by ASON didn't inhibit proliferation in LAK cells, but reduced the cytotoxicity for H22 and K562 cells significantly.
出处
《中华检验医学杂志》
CAS
CSCD
北大核心
2004年第5期327-329,共3页
Chinese Journal of Laboratory Medicine
基金
山东省科技厅重点课题资助项目(1995BB1CJA6)
