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目的:探讨核糖核酸酶抑制因子(RI)在卵巢癌裸鼠移植瘤治疗中的作用。方法:用重组质粒(PLNCX-RI)转化DH5-α感受态细菌,成功转化后以SDS碱裂解法大量制备纯化质粒DNA,同法处理空质粒载体(PLNCX)。通过琼脂糖电泳、紫外分光光度法鉴定R1基因提取的纯度和浓度。体外培养卵巢癌SKOV3细胞,取对数生长期细胞皮下注射裸鼠,建立卵巢癌裸鼠皮下移植瘤模型。对照组以空质粒载体(PLNCX)为治疗试剂,实验组以质粒DNA(PLNCX-RI)为治疗试剂进行皮下注射。比较两组裸鼠成瘤率、抑瘤率、成瘤时间及瘤体重量。免疫组织化学法分析两组RI基因的表达,HE染色对比观察微血管密度。结果:电泳检测出现了PLNCX-RI和PLNCX的条带。治疗结果显示注射RI基因的实验组与注射空质粒载体的对照组的肿瘤成瘤期分别为(16±3)天、(11±3)天,成瘤率分别为60.00%、80.00%,瘤组织重量分别为(0.27±0.06)g、(2.60±0.59)g,两组比较差异均有统计学意义(P<0.05)。实验组肿瘤抑制率为89.60%,RI基因表达水平较高,肿瘤组织血管密度低于对照组,差异均有统计学意义(P<0.01)。结论:RI抑制了卵巢癌裸鼠皮下移植瘤的生长,其机制可能为RI通过抑制Ang活性途径抑制了肿瘤组织的血管形成。
Objective: To investigate the role of ribonuclease inhibitor (RI) in the treatment of ovarian cancer xenografts in nude mice. Methods: The recombinant plasmid (PLNCX-RI) was used to transform DH5-α competent cells. After successful transformation, plasmid DNA was prepared by SDS alkaline lysis and PLNCX was treated in the same way. The purity and concentration of R1 gene were identified by agarose gel electrophoresis and ultraviolet spectrophotometry. Ovarian cancer SKOV3 cells were cultured in vitro, nude mice were subcutaneously injected with cells in logarithmic growth phase, and subcutaneous xenograft model of ovarian cancer was established. In the control group, empty plasmid vector (PLNCX) was used as the therapeutic reagent, and the experimental group was injected subcutaneously with plasmid DNA (PLNCX-RI) as the therapeutic reagent. The tumor formation rate, tumor inhibition rate, tumor formation time and tumor weight of two groups were compared. The expression of RI gene in the two groups was analyzed by immunohistochemistry, and the microvessel density was observed by HE staining. Results: The bands of PLNCX-RI and PLNCX were detected by electrophoresis. The results of the treatment showed that the tumorigenicity of the experimental group injected with RI gene and the control group injected with empty plasmid vector were (16 ± 3) days and (11 ± 3) days, respectively. The tumorigenic rates were 60.00% and 80.00% Tissue weights were (0.27 ± 0.06) g and (2.60 ± 0.59) g, respectively, with significant differences between the two groups (P <0.05). The tumor inhibition rate of the experimental group was 89.60%, the expression level of RI gene was higher, and the density of tumor tissue was lower than that of the control group (P <0.01). Conclusion: RI inhibits the growth of subcutaneous xenografts in ovarian cancer nude mice. The mechanism may be that RI inhibits the angiogenesis of tumor tissue by inhibiting the activity of Ang.