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目的:通过卵巢癌细胞株SKOV-3细胞增殖及动物荷瘤实验探讨siRNA沉默Her2/neu基因的表达效果。方法:靶向Her2/neu的siRNA经脂质体L ipofectAM INE2000介导转染到SKOV-3中,通过嘌呤霉素筛选得到稳定抑制Her2/neu基因表达的SKOV-3/siRNA细胞株,通过RT-PCR和免疫组化方法检测Her2/neu基因抑制效果,运用四甲基偶氮唑蓝(MTT)检测各组细胞增殖水平并将SKOV-3/siRNA细胞接种到裸鼠皮下检测荷瘤情况。结果:建立了稳定抑制Her2/neu基因表达的细胞株SKOV-3/siRNA,RT-PCR和免疫组化结果显示Her2/neu基因表达受到较强抑制,MTT检测结果显示SKOV-3/siRNA组细胞增殖较SKOV-3明显下调(P<0.05),其接种在裸鼠皮下形成的肿瘤生长速度明显减慢(P<0.05)。结论:靶向Her2/neu基因的siRNA可以抑制卵巢癌细胞株SKOV-3在体外和体内增殖,有望成为卵巢肿瘤治疗的新途径。
OBJECTIVE: To investigate the effect of siRNA silencing Her2 / neu gene expression in ovarian cancer cell line SKOV-3 cell proliferation and animal tumor-bearing experiments. Methods: The siRNA targeting Her2 / neu was transfected into SKOV-3 by LipofectAM INE2000. SKOV-3 / siRNA cell lines stably inhibiting the expression of Her2 / neu gene were obtained by puromycin screening. The inhibitory effect of Her2 / neu gene was detected by PCR and immunohistochemistry. The proliferation of each group was detected by MTT assay and the SKOV-3 / siRNA cells were inoculated subcutaneously in nude mice to detect tumor-bearing status. Results: The cell line SKOV-3 / siRNA which stably inhibited the expression of Her2 / neu gene was established. The expression of Her2 / neu gene was strongly inhibited by RT-PCR and immunohistochemistry. The results of MTT assay showed that SKOV- Compared with SKOV-3, the proliferation of SKOV-3 cells was significantly down-regulated (P <0.05). The growth of tumors in nude mice subcutaneously reduced significantly (P <0.05). Conclusion: siRNA targeting Her2 / neu gene can inhibit the proliferation of ovarian cancer cell line SKOV-3 in vitro and in vivo and is expected to become a new approach for the treatment of ovarian tumors.