黄芩苷通过上调miR-485-5p的表达抑制前列腺癌细胞的增殖

Baicalin inhibits the invasion of prostate cancer cells by up-regulating the expression of miR-485-5p

目的:从miRNAs的角度研究黄芩苷对前列腺癌细胞增殖的影响及其作用机制。方法:将不同浓度的黄芩苷作用22RV1细胞,应用MTT检测细胞的增殖情况,筛选最佳药物浓度;应用Real-Time PCR检测不同细胞系中miR-485-5p的表达量;检测黄芩苷处理22RV1细胞后miR-485-5p及JAK3 mRNA的表达;应用Western blot检测JAK3蛋白的表达;应用双荧光素酶基因系统验证miR-485-5p与JAK3的靶向作用关系;应用MTT法检测细胞的生存率。结果:筛选出最佳药物浓度为100μmol·L^(-1);22RV1细胞中miR-485-5p的表达量明显低于正常前列腺细胞;黄芩苷刺激细胞后miR-485-5p的表达上调;黄芩苷或miR-485-5p mimics可抑制前列腺癌细胞的JAK3 mRNA和蛋白的表达,miR-485-5p inhibitor可使JAK3 mRNA及蛋白的表达水平上调;黄芩苷或miR-485-5p均可抑制前列腺癌细胞的增殖。结论:黄芩苷通过上调miR-485-5p的表达,抑制前列腺癌细胞的增殖。

OBJECTIVE To observe the effect of baicalin on proliferation of prostate cancer cells and its mechanism from the view of miRNAs. METHODS Human prostate cancer cell line 22 RV1 was treated with different concentrations of baicalin. The proliferation of prostate cancer cell line 22 RV1 was determined by MTT. The best drug concentration was chosen. Real time PCR was used to examine the expression of miR-485-5 p in different cell lines,and the expressions of miR-485-5 p and JAK3 mRNA after treated by baicalin,and Western blot was used to examine the expression of JAK3 protein. Double luciferase gene system was used to verify the targeting relationship between miR-485-5 p and JAK3. The cell survival rate was detected by MTT. RESULTS The best drug concentration of 100 μmol·L^-1 was optimal,the expression of miR-485-5 p in 22 RV1 cells was significantly lower than that of normal prostate cells,expression of miR-485-5 p was up-regulated in Baicalin treated prostate cancer cells. Baicalin and miR-485-5 p could inhibit the expressions of JAK3 mRNA and protein in prostate cancer cells,expression of JAK3 mRNA and protein were up-regulated by miR-485-5 p inhibitor. Baicalin and miR-485-5 p could inhibit the proliferation of prostate cancer cells. CONCLUSION Baicalin can inhibit the proliferation of prostate cancer cells. The mechanism is related to the up-regulation of miR-485-5 p expression.