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目的观察UbcH10基因沉默对多柔比星抑制耐药乳腺癌细胞株MCF-7/ADR体内成瘤作用的影响。方法采用慢病毒系统在MCF-7/ADR细胞中进行UbcH10基因沉默。将基因沉默后的肿瘤细胞及对照肿瘤细胞接种至裸鼠皮下,通过裸鼠尾静脉连续给予多柔比星或生理盐水2周并停药1周,测量肿瘤体积,分析UbcH10沉默对多柔比星抑制MCF-7/ADR体内成瘤作用的影响。采用蛋白质印迹分析检测瘤体中的UbcH10及BCL-2蛋白含量,分析UbcH10与肿瘤细胞化疗敏感性之间的调控关系。结果通过慢病毒实验系统成功建立UbcH10基因沉默细胞株MCF-7/ADR UbcH10-RNAi。经过3周给药处理,多柔比星组肿瘤体积与对照组比较差异无统计学意义,肿瘤抑制率为4.16%;基因沉默+多柔比星组肿瘤体积与对照组差异有统计学意义(P<0.05),肿瘤抑制率为41.8%。瘤体内蛋白含量检测结果显示,对照组、多柔比星组、基因沉默+多柔比星组UbcH10蛋白表达量分别为0.81±0.16、0.78±0.12、0.18±0.04,基因沉默+多柔比星组其余两组比较差异均有统计学意义(P<0.05);BCL-2蛋白表达量组间差异与UbcH10一致,二者之间具有正相关性。结论UbcH10基因沉默可以显著增强多柔比星抑制耐药乳腺癌细胞MCF-7/ADR的体内成瘤作用。
Objective To investigate the effect of UbcH10 gene silencing on in vivo tumorigenesis of doxorubicin-resistant breast cancer cell line MCF-7 / ADR. Methods The lentivirus system was used to silence UbcH10 gene in MCF-7 / ADR cells. Tumor cells and control tumor cells after gene silencing were inoculated subcutaneously into nude mice, and doxorubicin or normal saline was continuously administered to the caudal vein of nude mice for 2 weeks and discontinued for 1 week. Tumor volume was measured, and the effect of UbcH10 silencing on multi- Star suppression of MCF-7 / ADR in vivo tumorigenic effect. The protein content of UbcH10 and BCL-2 in the tumor was detected by Western blot analysis, and the relationship between UbcH10 and chemosensitivity of tumor cells was analyzed. Results The UbcH10 gene silencing cell line MCF-7 / ADR UbcH10-RNAi was successfully established by lentiviral experiment system. After 3 weeks of treatment, the tumor volume in doxorubicin group was not significantly different from that in control group, the tumor inhibition rate was 4.16%. The tumor volume in gene silencing + doxorubicin group was significantly different from that in control group P <0.05), the tumor inhibition rate was 41.8%. The detection results of the protein content in the tumor showed that the expression of UbcH10 protein in the control group, doxorubicin group and gene silencing + doxorubicin group were 0.81 ± 0.16, 0.78 ± 0.12 and 0.18 ± 0.04, respectively. Gene silencing + doxorubicin There was significant difference between the other two groups (P <0.05). The difference of BCL-2 protein expression between the two groups was consistent with that of UbcH10. There was a positive correlation between them. Conclusion UbcH10 gene silencing can significantly enhance the in vivo tumorigenesis of doxorubicin in inhibiting the MCF-7 / ADR-resistant breast cancer cells.