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目的:研究SO-Rb50细胞系瘤细胞Rb基因突变的动态变化。方法:1.用 Southern blot杂交法分析 SO-Rb50细胞系第327代瘤细胞DNA。2.用PCR-SSCP法对SO-Rb50细胞系第415代及第713代瘤细胞的Rb基因27个外显子和1个启动子逐个筛查。3.将SO-Rb50细胞系的第775代克隆出3个细胞株:MC2、MC3及MC4。用PCR-SSCP-HA法对MC2、MC3及MC4第11代细胞和MC3第138代细胞的Rb基因的27个外显子逐个筛查。结果:SO-Rb50细胞系第327代细胞DNA缺失3.5Kb、2.9Kb及1.0Kb的三条带,证明SO-Rb50细胞系瘤细胞Rb基因缺失。第415代瘤细胞Rb基因第23外显子少一条单链;第713代第25外显子发生新的突变。SO-Rb50细胞系第775代的3个克隆株MC2、MC3及MC4的第11代细胞,只有MC4第24外显子突变;而MC3第138代第24外显子突变,提示MC3经多次传代后第24外显子发生新突变。结论:SO-Rb50细胞系在长期培养传代过程中,瘤细胞的Rb基因突变有动态变化。眼科学报2001;17:111～113。
Objective: To study the dynamic changes of Rb gene mutation in SO-Rb50 cell line. Methods: 1. Southern blot hybridization analysis SO-Rb50 cell line 327 tumor cells DNA. The 27 exons and one promoter of Rb gene in the 415th and the 713th generation of SO-Rb50 cell line were screened by PCR-SSCP. 3. The 775th generation of SO-Rb50 cell line cloned three cell lines: MC2, MC3 and MC4. 27 exons of Rb gene of MC2, MC3 and MC4 11th generation cells and 138th generation MC3 cells were screened by PCR-SSCP-HA. Results: Three bands of 3.5Kb, 2.9Kb and 1.0Kb were deleted in DNA of 327th generation in SO-Rb50 cell line, which proved that the Rb gene of SO-Rb50 cell line was deleted. The 415th tumor cell Rb gene 23 exon less than a single chain; 713 generation 25th exon new mutation. Only the exon 24 of MC4 was mutated in the 11th generation of MC2, MC3 and MC4 in the 775th generation in SO-Rb50 cell line, and the mutation of exon 24 in 138th generation of MC3, A new mutation occurs in the 24th exon after passage. Conclusion: The mutation of Rb gene in tumor cells in SO-Rb50 cell line has a dynamic change during the passage of long-term culture. Journal of Ophthalmology 2001; 17: 111 ~ 113.