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目的:研究微小RNA-16(miR-16)调控靶基因CCND1及其蛋白表达产物-细胞周期相关蛋白D1(cyclinD1)在卵巢肿瘤细胞系SKOV3及其耐药株SKOV3/DDP中的表达,并初步探讨其逆转卵巢癌耐药的可能机制。方法:脂质体Lipofectamine2000介导miR-16成体模拟物(mimics)及其阴性对照片段(NC)成功转染人卵巢癌细胞顺铂耐药株SKOV3/DDP;流式细胞仪检测转染前后细胞周期的变化;Western blot法检测转染前后cyclinD1表达的变化。结果:(1)成功转染miR-16后,SKOV3/DDP细胞系G1期细胞所占比例显著升高(P<0.01);(2)SKOV3/DDP细胞的cyclinD1蛋白相对表达量显著高于SKOV3细胞;转染miR-16后,cyclinD1相对表达量显著下降(P<0.01)。结论:miR-16很可能通过下调靶基因CCND1及cyclinD1蛋白的表达,阻止细胞由G1期向S期转换,抑制肿瘤细胞无限增殖与生长。
OBJECTIVE: To study the expression of CCND1, a target gene of microRNA-16 (miR-16), and its cyclinD1 protein in ovarian tumor cell line SKOV3 and its drug-resistant cell line SKOV3 / DDP To explore its possible mechanism of reversing the drug resistance of ovarian cancer. METHODS: Human ovarian cancer cell line SKOV3 / DDP was successfully transfected with mimics and its negative control (mimics) mediated by Lipofectamine 2000. The cells were detected by flow cytometry before and after transfection The changes of cyclinD1 expression before and after transfection were detected by Western blot. Results: (1) The percentage of G1 phase cells in SKOV3 / DDP cell line was significantly increased after miR-16 was successfully transfected (P <0.01). (2) The relative expression of cyclinD1 in SKOV3 / DDP cells was significantly higher than that in SKOV3 After transfection with miR-16, the relative expression of cyclinD1 was significantly decreased (P <0.01). Conclusion: miR-16 is likely to inhibit the proliferation and growth of tumor cells by down-regulating the expression of target genes CCND1 and cyclinD1, and preventing cells from transforming from G1 phase to S phase.