TRAIL诱导肿瘤细胞凋亡及其与FHIT基因突变关系的初步研究

目的:分析原核重组表达的人可溶性TRAIL分子诱导多种肿瘤细胞的凋亡,并观察凋亡作用与FHIT基因突变的关系.方法:观察人可溶性TRAIL对28株不同来源的肿瘤细胞和正常细胞的直接细胞毒作用,RT-PCR法分析部分细胞的FHIT基因缺失突变情况.结果:1～100 mg/L的重组人可溶性T RAIL蛋白即可诱导19株细胞(包括人肿瘤细胞株)如1990(胰腺导管癌)、MCF-7(乳腺癌)、A5 4 9(肺癌)、U251(星形胶质瘤)、HepG-2(肝细胞癌)、M85(胃癌)、CNE-2(鼻咽癌)、Hep-2( 喉癌)、AT-29(结肠癌)、3AO(卵巢癌)和B16-MB(黑色素瘤),髓性恶性细胞如Jurkat、K56 2、U937、6T-CEM,鼠源性S180(肉瘤)、Ehrlich(腹水瘤)和Lewis(肺癌),以及人内皮细胞株ECV304等发生凋亡;而其他9株细胞,如SK-N-SH(人神经母细胞瘤)、8898(人胰腺导管癌)、HeLa(人宫颈癌)、SMMU7721(人肝癌)、HL60(人白血病)、COS-7(猴肾)、人成纤维细胞、L929(鼠成纤维细胞)、Wish(人羊膜细胞)等需大于100 mg/L的TRAIL才能使其凋亡;观察到上述部分对TRAIL敏感的人源细胞的FHIT基因有缺失突变,而不敏感的人源细胞FHIT 基因完整.结论:原核表达的人可溶性TRAIL具有明显诱导多种肿瘤细胞凋亡的活性,其机制可能与FHIT基因的缺失有关.

Relationship between induction of apoptosis by TRAIL and FHIT gene mutation in various tumor cell lines

Objective:To investigate the induction of apoptosis by hum an soluble TRAIL expressed in prokaryote and its relationship with FHIT gene mutation in various tumor cell lines.Methods:TRAIL was observed for its in vitro direct cytostatic or cytotoxic effects on 2 8cell lines from colon,lung,breast,kidney,brain,skin,myoloblastic cancer and norm al tissues.In some cell lines,the deletions of FHIT gene were detected by RT- PCR.Results:The concentration from 1to 10 0 mg/ L had a m arked apoptosis in 19of 2 8cell lines including human 1990 (pancreatic carcinom a) ,MCF- 7(breast cancer) ,A5 4 9 (lung cancer) ,U 2 5 1(astrogliom a) ,Hep G- 2 (hepatocellular carcinoma) ,M85 (gastric cancer) ,CNE- 2 (nasopharyngeal carcinom a) ,Hep- 2 (laryngeal carcinom a) ,AT- 2 9(colon cancer) ,3AO (ovarian carcinoma) and B16 - MB (m elanoma) ; myeloneoplastic included Jurkat,K5 6 2 ,U 937and 6 T- CEM;murine tum or cells included S180 (sarcom a) ,Ehrlich (ascites carcinom a) and L ewis (lung cancer) ;and also to human endothelial cell line ECV30 4 .The concentration over 10 0 mg/ L was needed for induction of apoptosis to other cell lines including SK- N - SH (human neuroblastoma) ,8898(human pancreatic carcinom a) ,He L a (human cervical carcinoma) ,SMMU772 1(hum an hepatocellular carcinoma) ,HL 6 0 (human promyelocytic leukemia) ,COS- 7(monkey renal) ,human and mouse (L 92 9) fibroblast cells,and Wish(hum an amniotic cell) .The deletion of FHIT was seen in some employed cell lines,while there were no mutations of FHIT gene for those insensitive cells. Conclusion:Human soluble TRAIL has a distinguished apoptosis to various transformed cell lines,and this effect seems to be related with the deletion of FHIT gene in sensitive cells.