乳腺癌细胞p16基因的研究

目的 探讨Ｐ１６基因在乳腺癌中的突变形式和频率以及有效的检测手段。方法 采用细胞２,反复贴壁法纯化人乳腺癌细胞、经多聚酶链反应－单链构象多态性（ＰＣＲ－ＳＳＣＰ）分析技术。对１４例纯化乳腺癌细胞标本,１４例新鲜未纯化标本和２７例石蜡包埋标本及相应的癌旁乳腺组织标本的这６基因突变进行了对比性研究。结果 纯化的癌细胞组Ｐ１６基因突变率为４２．９％,而新鲜未纯化细胞组、石蜡包埋细胞组及癌旁乳腺细胞组的Ｐ

p16 gene deletion and point mutation in highly purified fresh breast neoplasm

OBJECTIVE: To study the form and frequency of p16 gene deletion and point mutation in human breast neoplasm and the effective detection. METHODS: Cell culture was made using repeat-adhering method to purify human breast neoplasm cells and PCR-SSCP analysis in 14 highly purified fresh breast neoplasm, 14 fresh none-purified specimens, 27 paraffin-embedded specimens and corresponding normal tissues beside neoplasms. RESULTS: The rate of p16 gene deletion and point mutation was 42.9% (6/14) in the highly purified group, in which exon 1st - 3rd, showed homozygous deletion (5 cases) and exon 2nd showed mutation (1 case). The rate of p16 gene mutation in the other three groups was 7.1%, 7.4% and 4.9%. The detection rate of mutation was significantly higher in the highly purified group than in the other three groups (P < 0.01), but it was not significantly different in the other three groups (P > 0.05). CONCLUSIONS: p16 gene deletion and point mutation exist in primary breast cancer. Homozygous deletion and small fragment deletion may be an important mechanism of inactivation in human breast neoplasm. Purified cancer cells will enhance the detection rate.