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研究了卵巢癌中高表达的新基因spindlin1的亚细胞定位及其对小鼠NIH3T3细胞生物学行为的影响 ,并探讨了人spindlin1基因的生物学功能 .采用脂质体转染法将构建的融合蛋白表达载体 pEGFP N1/pEGFP N1 spindlin1瞬时转染COS 7细胞 ,观察spindlin1基因的亚细胞定位 ;同时稳定转染NIH3T3细胞 ,经G4 18抗性筛选后 ,用RT PCR筛选并鉴定表达spindlin1的稳定细胞株 ;通过细胞增殖活性、流式细胞检测、软琼脂克隆形成、迁移实验及裸鼠成瘤等对转染细胞的生物学行为进行检测 .结果表明 ,spindlin1定位于胞核 ;并促进NIH3T3细胞的增殖 ,使其处于G2 /M期的细胞比例显著增加 ;同时证实过表达spindlin1的NIH3T3细胞不仅具明显的克隆形成及迁移能力 ,而且能使裸鼠成瘤 .由此我们得出结论 :spindlin1基因过表达可促使NIH3T3细胞发生恶性转化 ,提示spindlin1可能是一种与肿瘤形成相关的原癌基因 .
We investigated the subcellular localization of spindlin1, a novel gene overexpressed in ovarian cancer, and its effect on the biological behavior of mouse NIH3T3 cells, and explored the biological function of human spindlin1 gene.The fusion protein The expression vector pEGFP N1 / pEGFP N1 spindlin1 was transiently transfected into COS 7 cells to observe the subcellular localization of spindlin1 gene. At the same time, the NIH3T3 cells were stably transfected. After being screened by G4 18, the stable cell lines expressing spindlin1 ; The biological behavior of transfected cells was detected by cell proliferation activity, flow cytometry, soft agar colony formation, migration assay and nude mouse tumorigenesis.The results showed that spindlin1 localized in the nucleus and promoted the proliferation of NIH3T3 cells , So that the proportion of cells in G2 / M phase significantly increased; at the same time, it was confirmed that overexpression of spindlin1 in NIH3T3 cells not only has obvious clonogenicity and migration ability, but also can make nude mice tumor.We concluded that spindlin1 gene Expression can promote the malignant transformation of NIH3T3 cells, suggesting that spindlin1 may be a tumor-associated proto-oncogene.