CD44反义寡核苷酸对肿瘤细胞表面抗原和对CD3AK杀伤敏感性的调节作用

Effects of CD44 antisense oligo deoxyribonucleotides(ASODN) on the expression of surface antigen of the tumor cells and susceptibility of target cells to CD3AK attack

目的:观察CD44反义寡核苷酸调节人低分化黏液腺胃癌MGC80-3细胞Fas分子和凋亡抵抗基因bc l-2的表达水平,提高免疫效应细胞杀伤敏感性的作用和机制。方法:RT-PCR法检测CD44、bc l-2 mRNA的表达水平。MTT法检测CD3AK杀伤活性。流式细胞术检测细胞表面CD44及Fas、FasL蛋白表达水平。结果:CD44反义寡核苷酸(1.6μmol/L)处理后,明显地抑制MGC80-3细胞CD44 mRNA和蛋白表达水平,在CD44配体低分子量透明质酸(HA)存在下,使MGC80-3表面下调的Fas分子显著增高,表达率从6.69%提高到16.81%(P<0.01)。CD3AK对反义寡核苷酸处理的MGC80-3细胞杀伤活性显著增高,并呈效靶比依赖效应(P<0.01)。MGC80-3细胞bc l-2 mRNA的相对表达定量值由1.06降至0.32。结论:CD44反义寡核苷酸可通过抑制MGC80-3细胞CD44 mRNA和蛋白表达,上调Fas分子的表达,下调凋亡抵抗基因bc l-2的表达,并提高其对免疫效应细胞的杀伤敏感性。

ObJective：To study the role and mechanism of CD44 ASODN on expression of surface molecule of tumor cells and susceptibility of target cells to immune cells attack. Methods：CIM4 mRNA and bcl-2 mRNA were examined by RT-PCR. The effect of CD44 ASODN on MGCS0-3 ceils sensitivity to lysis by CD3AK effector cells was examined by MTY assay. The expression of CD44 and Fas was examined by FACS. Results：Treated with CD44 ASODN（ 1.6 μmol/L）, there was a marked reduction of expression of CD44 protein and mRNA on MGC80-3 cells. Expression ratio of Fas was enhanced from 6. 69% to 16. 81% （ P 〈 0. 01 ）. Expression of bcl-2 mRNA obviously reduced. After treated with CD44 ASODN,the susceptitility of MGC80-3 cells to lysis by CD3AK was enhanced obviously（ P 〈 0. 01 ） at the different ratio of effeetor to target and was time and dose dependent. Concluslon：CD44 ASODN can block and inhibit expression of CD44 mRNA and proteln, enhance expression of Fas, reduce expression of bcl-2, which agument sensitivity of MGC80-3 ceils to lysis by CD3AK effector cells.