反义RNA抑制人黑色素瘤细胞表面粘附分子CD44的表达及抑瘤效应

Inhibition of CD44 Expression and Tumorigenicity of Human Melanoma Cell Line by Antisense RNA

将细胞表面粘附分子ＣＤ４４Ｓ的ｃＤＮＡ反向插入到真核细胞表达载体ｐＭＡＭｎｅｏ－ＣＡＴ和ＭＭＴＶ－ＬＴＲ启动子下游，构成ＣＤ４４Ｓ的反义ＲＮＡ载体．将其用电击法导入ＣＤ４４＋的人黑色素瘤细胞系ＨＭＭ２３９，转录出的反义ＲＮＡ能不同程度地抑制ＨＭＭ２３９表面ＣＤ４４的表达．ＣＤ４４的表达被抑制后，瘤细胞与透明质酸的结合力下降，细胞的体外生长速率不受影响．将其接种裸鼠皮下。

CD44 is a widely distribulated cell surface glycoprotein involved in cell adhesion,lymphocyte activation and lymphocyte homing.It is also implicated in the regulation of tumor growth and metastasis,but the mechanism remains unclear.It was investigated that the role of CD44 on human melanoma cell line HMM239 with antisense RNA technique.The antisense RNA plasmid vector was constructed and electroporated into HMM239 cells.FACS analysis revealed that the expression of CD44 on the surface of transfected HMM239 cells could be blocked by antisense RNA.When CD44 on the cells was almost completely blocked,the cells lost the binding activity to HA.When the tumor cells were injected subcutaneously into a thymic nude mice,the tumorigenicity of CD44 negative tumor cells decreased greatly as compared with CD44 positive tumor cells,but there was no difference in their growth rate in vitro.Therefore,CD44 ligand interaction in vivo may be necessary for tumor growth.