RNA干扰对XWLC-05肺癌细胞水通道蛋白3表达的影响

Knockdown the expression of aquaporin-3(AQP3) in XWLC-05 human lung cancer cells by small interfering RNA

目的构建靶向水通道蛋白3(AQP3)的siRNA表达载体,探讨其对XWLC-05肺癌细胞系AQP3表达的影响。方法构建AQP3特异性siRNA表达载体和对照组表达载体,以脂质体法转染XWLC-05肺癌细胞,用RT-PCR检测AQP3 mRNA的表达,用Western blot方法检测APQ3蛋白的表达。结果成功构建靶向AQP3的siRNA表达载体1和2,分别命名为pAQP3-siRNA1、pAQP3-siRNA2。转染48 h后pAQP3-siRNA1和pAQP3-siRNA2转染组AQP3 mRNA的表达量分别是对照组的65%和79%,组间比较有统计学差异(P<0.05);pAQP3-siRNA1和pAQP3-siRNA2转染后的蛋白表达量分别是对照组的53%和73%,pAQP3-siRNA1干扰效果明显高于pAQP3-siR-NA2(P<0.05)。结论 AQP3特异性siRNA能够有效地抑制XWLC-05肺癌细胞系AQP3的表达。

Objective To construct the siRNA expression vector targeting AQP3 and to evaluate its ability to down-regulate the expression of AQP3 in XWLC-05 human lung cancer cells. Methods Two siRNA expression vectors, pAQP3- siRNA1, pAQP3-siRNA2 targeting AQP3, were constructed. After pAQP3-siRNA1 and pAQP3-siRNA2 were transfected into XWLC-05 lung cancer cells, the levels of mRNA and protein expressions in XWLC-O5 human lung cancer ceils were determined by RT-PCR and Western blot analysis respectively. Results Two siRNA expression vectors targeting AQP3 were constructed successfully, named pAQP3-siRNA1 and pAQP3-siRNA2 respectively. Forty-eight hours after transfection, level of AQP3-mRNA was decreased significantly in AQP3-aiRNA1 XWLC-05 ceils （65% of that in control group） compared with AQP3-siRNA2 XWLC-05 cells （79% of that in control group ） （P 〈 0.05 ）. The level of protein expression of AQP3 in AQP3-siRNA1 XWLC-05 cells decreased significantly （53% of that in control group ） compared with AQP3- siRNA2 XWLC-05 cells （73% of that in control group） （ P 〈 0.05）. Conclusions AQP3 specific siRNA can significantly and specifically down-regulate the expression of AQP3 in XWLC-05 cells.