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目的了解体外转录法合成的小干扰 RNA(siRNA)沉默 MAT1基因对胰腺癌的体内体外抗癌效应。方法用 siPORT-脂质体(lipid)中性阳离子脂质体转染由体外转录法合成、Cy3荧光标记的双链 siRNA 人胰腺癌 BxPC3细胞,观察对胰腺癌细胞生长和细胞周期曲线的影响,检测 MAT1基因被 siRNA 沉默后 mRNA 和蛋白质的表达水平。在裸鼠胰腺癌皮下移植瘤瘤体内直接注射 siRNA观察抑瘤效应。结果合成的4条 MAT1-siRNA 序列中有一半可显著抑制 BxPC3细胞的生长。体外转染 MAT1-siRNA 72 h 后,BxPC3细胞生长抑制率达34.9%(P<0.01);83.9%的 BxPC3细胞滞留于G_0/G_1期,而空白对照组仅59.86%(P<0.01)。体外转染 MAT1-siRNA 48 h 后 MAT1-mRNA 水平下调了80.12%,72 h 后 MAT1蛋白质水平下调了50.12%,与各对照组相比差异均有统计学意义(均 P<0.01)。MAT1-siRNA 注射组裸鼠移植瘤重量和体积均明显低于对照组(均 P<0.05),抑瘤率达42.53%。结论 siRNA 介导的 MAT1基因沉默在体外实验中显著抑制了胰腺癌细胞的生长,在体内实验中对裸鼠胰腺癌皮下移植瘤产生明显的抑瘤效应。
Objective To investigate the in vitro antitumor effect of MAT1 gene silencing small interfering RNA (siRNA) synthesized by in vitro transcription on pancreatic cancer in vitro and in vivo. Methods The human pancreatic cancer BxPC3 cells were transfected with siPORT-liposome-neutral cationic liposomes by Cy3 fluorescent-labeled double-stranded siRNA in vitro and the effects on the growth and cell cycle curve of pancreatic cancer cells were observed. The mRNA and protein levels of MAT1 gene were silenced by siRNA. Injecting siRNA directly into the subcutaneously transplanted tumor of pancreatic cancer in nude mice to observe the anti-tumor effect. Results Half of the 4 MAT1-siRNA sequences synthesized could significantly inhibit the growth of BxPC3 cells. The growth inhibition rate of BxPC3 cells was up to 34.9% (P <0.01) after transfected with MAT1-siRNA for 72 h in vitro. 83.9% of BxPC3 cells remained in G_0 / G_1 phase compared with 59.86% of the blank control group (P <0.01). The MAT1-mRNA level was down-regulated by 80.12% at 48 h after MAT1-siRNA transfection in vitro. After 72 h, the MAT1 protein level was reduced by 50.12%, which was significantly different from the control group (all P <0.01). The tumor weight and volume of MAT1-siRNA injected group were significantly lower than that of the control group (all P <0.05), and the tumor inhibition rate was 42.53%. Conclusion siRNA-mediated MAT1 gene silencing significantly inhibits the growth of pancreatic cancer cells in vitro, and has obvious anti-tumor effect on subcutaneously transplanted pancreatic cancer in nude mice in vivo.