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目的探讨应用小分子干扰RNA(small interfering RNA,siRNA)沉默抑癌基因LKB1对人乳腺癌细胞MDA-MB-231中Hedgehog信号通路相关因子的表达及人乳腺癌裸鼠移植瘤模型的肿瘤生长的影响。方法构建LKB1基因siRNA质粒LKB1-siRNA;建立LKB1表达抑制的MDA-MB-435细胞模型;裸鼠乳晕皮下接种,建立人乳腺癌裸鼠移植瘤动物模型;成瘤后,观察肿瘤体积变化、裸鼠生存时间;并用Western印迹法检测瘤组织中LKB1和Hedgehog信号通路中信号肽Shh、Sufu、膜受体Ptch、Smo、转录因子Gli1、Hip蛋白表达的变化。结果 LKB1-siRNA质粒组裸鼠的肿瘤体积明显增长(P<0.05);肿瘤内LKB1基因表达水平明显下降,而Hedgehog信号通路相关因子Shh、Gli1、Ptch、Smo的表达升高,Hedgehog信号通路抑制因子Sufu、Hip表达下降。结论 LKB1基因siRNA能够明显抑制人乳腺癌裸鼠移植模型的LKB1基因的表达,上调Hedgehog信号通路相关因子的表达,促进肿瘤生长。LKB1基因和Hedgehog信号通路在乳腺癌细胞中呈现负相关表达。
Objective To investigate the expression of Hedgehog signaling pathway-related factors in human breast cancer cell line MDA-MB-231 and the tumor growth of human breast cancer xenografts in nude mice using small interfering RNA (siRNA) silencing of tumor suppressor gene LKB1 influences. Methods LKB1 siRNA siRNA plasmid LKB1-siRNA was constructed. The MDA-MB-435 cell model was established to inhibit the expression of LKB1. The nude mice were subcutaneously inoculated with the areola and the tumor model was established. The survival time of rats was observed. The changes of signal peptide Shh, Sufu, membrane receptors Ptch, Smo, transcription factors Gli1 and Hip in LKB1 and Hedgehog signaling pathways were detected by Western blotting. Results The tumor volume of LKB1-siRNA plasmid group was significantly increased (P <0.05). The expression of LKB1 gene in tumor was significantly decreased, while the expression of Shh, Gli1, Ptch and Smo in Hedgehog signaling pathway was increased and Hedgehog signaling pathway was inhibited Factor Sufu, Hip expression decreased. Conclusion LKB1 gene siRNA can significantly inhibit the expression of LKB1 gene in human breast cancer xenograft model, up-regulate the expression of Hedgehog signaling pathway-related factors and promote tumor growth. The LKB1 gene and the Hedgehog signaling pathway are negatively correlated in breast cancer cells.