摘要
目的 探讨mi R-339-3P靶向六磷酸肌醇激酶2(IP6K2)对前列腺癌细胞增殖、凋亡、侵袭和上皮间充质转化(Epithelial Mesenchymal Transition EMT)的作用及其机制。方法 应用实时荧光定量PCR(qRT-PCR)分别检测前列腺癌组织和癌旁组织中mi R-339-3P和IP6K2的表达水平。利用细胞转染技术向人前列腺癌细胞PC-3分别转染mi R-339-3P-mimics、mi R-339-3P inhibitors、si-IP6K2(抑制表达)和PcDNA-3.1(+)-IP6K2(过表达),CCK-8检测细胞增殖情况,流式细胞仪检测细胞凋亡及细胞周期情况,细胞划痕检测细胞迁移能力,Western blot检测靶基因IP6K2和EMT相关因子的蛋白表达水平。采用双荧光素酶报告基因实验验证mi R-339-3P和IP6K2的靶向结合关系。结果qRT-PCR的结果显示,前列腺癌组织中mi R-339-3P表达水平低于癌旁组织,IP6K2表达水平高于癌旁组织,差异有统计学意义(P<0.05)。miR-339-3P-mimics抑制IP6K2表达可以显著抑制PC-3细胞增殖与迁移,促进细胞凋亡,Twist1、N-cadherin和Fibronectin蛋白表达降低,E-cadherin蛋白表达增加。miR-339-3P inhibitors过表达IP6K2可以显著促进PC-3细胞增殖、迁移和EMT,并抑制细胞凋亡。结论 过表达mi R-339-3P通过靶向抑制IP6K2基因表达,从而抑制前列腺癌细胞增殖和迁移能力,抑制EMT发生并促进细胞凋亡。
Objective To investigate the effect and the mechanism of mi R-339-3P targeting inositol hexaphosphate kinase 2(IP6K2) on proliferation, apoptosis, invasion and epithelial mesenchymal transformation(EMT) of prostate cancer cells. Methods The expression levels of mi R-339-3P and IP6K2 in prostate cancer tissues and adjacent tissues were detected by real-time fluorescence quantitative PCR(qRT-PCR). Mi R-339-3P-mimics, mi R-339-3P inhibitors, si-IP6K2(inhibited expression) and PcDNA-3. 1(+)-IP6K2(overexpression) were transfected into human prostate cancer cell line PC-3 by the cell transfection technique. CCK-8 was used to detect the cell proliferation;flow cytometry was used to detect the apoptosis and the cell cycle;cell scratch was used to detect the cell migration;and Westernblot was used to detect the protein expression of target gene IP6K2 and EMT related factors. Double luciferase reporter gene assay was used to verify the targeted binding relationship between mi R-339-3P and IP6K2. Results The results of qRT-PCR showed that the expression level of mi R-339-3P in prostate cancer tissues was significantly lower than that in the adjacent tissues, while the expression level of IP6K2 in prostate cancer tissues was significantly higher than that in the adjacent tissues(P<0. 05). MiR-339-3P-mimics inhibition of IP6K2 expression could significantly inhibit the proliferation and migration of PC-3 cells and promote apoptosis. The expression of Twist1, N-cadherin and Fibronectin protein wasdecreased, while the expression of E-cadherin protein wasincreased. On the contrary, miR-339-3P inhibitors over-expression of IP6K2 could significantly promote the proliferation, migration and EMT of PC-3 cells, and itcouldinhibit apoptosis. Conclusion over-expression of miR-339-3P can probably inhibit the proliferation and migration of prostate cancer cells, and it can inhibit the occurrence of EMT and promote apoptosis by targeting inhibition of IP6K2 gene expression.
作者
王文光
王玉杰
侯亚坤
宋鸿文
马军
阿斯木江·阿不拉
WANG Wenguang;WANG Yujie;HOU Yakun;SONG Hongwen;MA Jun;Asimujiang Abula(Center of Urology,The First Affiliated Hospital of Xinjiang Medical University/Xinjiang Clinical Research Center for Urogenital Diseases,Urumqi 830054,China)
出处
《新疆医科大学学报》
CAS
2022年第12期1367-1373,共7页
Journal of Xinjiang Medical University
基金
新疆维吾尔自治区自然科学基金(2019D01C314)。
