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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 siRNA expression vector targeting aquaporin 3 (AQP3) and investigate its effect on the expression of AQP3 in XWLC-05 lung cancer cell line. Methods AQP3 specific siRNA expression vector and control group were constructed and transfected into XWLC-05 lung cancer cells by lipofectamine 2000. The expression of AQP3 mRNA was detected by RT-PCR and the expression of APQ3 protein was detected by Western blot. Results The AQP3-targeting siRNA expression vectors 1 and 2 were successfully constructed and named as pAQP3-siRNA1 and pAQP3-siRNA2 respectively. After 48 h of transfection, the expression of AQP3 mRNA in pAQP3-siRNA1 and pAQP3-siRNA2 transfected groups was 65% and 79% of the control group, respectively, with statistical significance (P <0.05); pAQP3-siRNA1 and pAQP3- siRNA2 transfected protein expression levels were 53% and 73% of the control group, pAQP3-siRNA1 interference was significantly higher than pAQP3-siR-NA2 (P <0.05). Conclusion AQP3-specific siRNA can effectively inhibit the expression of AQP3 in XWLC-05 lung cancer cell line.