siRNA沉默细胞外信号调节激酶1/2基因协同甲亢宁胶囊对大鼠甲状腺细胞-5细胞增殖的影响

摘要

目的：通过甲亢宁胶囊（以下简称“甲亢宁”）协同siRNA干扰大鼠甲状腺细胞-5（fisher rat thyroid cell line-5，FRTL-5）中细胞外信号调节激酶（ERK）1/2基因，观察并探讨甲亢宁对FRTL-5细胞RNA干扰ERK1/2基因表达及对FRTL-5细胞增殖的影响。方法将生长状态良好的FRTL-5细胞分为空白组、阴性对照组、甲亢宁组、siRNA组、甲亢宁协同组。利用siRNA沉默及甲亢宁协同干扰FRTL-5细胞的ERK1/2基因， RT-PCR检测ERK1/2 mRNA表达，Western blot检测ERK1/2、p-ERK1/2蛋白表达，CCK8法检测细胞增殖。结果 RT-PCR检测结果显示，空白组和阴性对照组ERK1/2的mRNA表达无明显差异（P＞0.05）；与空白组比较，siRNA组和甲亢宁组FRTL-5细胞ERK1/2 mRNA表达明显下降（P＜0.01）；与空白组、阴性对照组比较，甲亢宁协同组ERK1/2 mRNA表达明显下降（P＜0.01）；Western blott检测结果显示，与空白组、阴性对照组比较，甲亢宁组、siRNA组、甲亢宁协同组p-ERK1/2、ERK1/2蛋白表达相对量明显下降（P＜0.01）。转染24、48 h后，CCK8法检测结果显示，甲亢宁组、siRNA组、甲亢宁协同组细胞增殖受抑制，与空白组、阴性对照组比较差异有统计学意义（P＜0.01）。结论甲亢宁对 ERK1/2磷酸化起阻断作用，ERK1/2基因被沉默后，对甲状腺细胞增殖有抑制作用，甲亢宁可协同ERK1/2-siRNA抑制FRTL-5细胞增殖。%ObjectiveTo observe and explore the effects ofJiakangning Capsules on the expression of ERK1/2 gene and proliferation of FRTL-5 by studying the effects ofJiakangning Capsules collaborated with siRNA on interfering ERK1/2 gene in FRTL-5.Methods FRTL-5 cells in good conditions were divided into control group, negative control group,Jiakangning Capsules collaborated with siRNA group, siRNA interference group, and Jiakangning Capsules group. RT-PCR was performed to detect the expression of ERK1/2 gene in mRNA levels in FRTL-5; Western blotting was performed to detect the expressions of ERK1/2 and p-ERK1/2; CCK8 method was used to detect cell proliferation.Results RT-PCR results showed that the ERK1/2 mRNA expression of the control group and negative control group were of no significant difference (P>0.05); compared with the control group, siRNA interference group andJiakangning Capsules group could inhibit ERK1/2 gene mRNA expression in FRTL-5 (P<0.01). Compared with the control group and negative control group, the ERK1/2 mRNA expression of Jiakangning Capsules collaborated with siRNA group decreased obviously (P<0.01). Compared with the control group and negative control group, the expression of p-ERK1/2, ERK1/2 ofJiakangning Capsules group, Jiakangning Capsules collaborated with siRNA group, and siRNA interference group decreased obviously (P<0.01). At the time of 24 h and 48 h after transfection, according to the results of CCK8, compared with the control group and negative control group, the cell proliferation ofJiakangning Capsules group, Jiakangning Capsules collaborated with siRNA group, and siRNA interference group was inhibited (P<0.01).Conclusion Jiakangning Capsules have blocking effect on the phosphorylation of ERK1/2. After ERK1/2 gene was silenced, the thyroid cell proliferation was inhibited. Jiakangning Capsules can collaborate with ERK1/2-siRNA to inhibit FRTL-5 cell proliferation.