微小RNA-375对鼻咽癌细胞侵袭迁移及JAK2/STAT3信号通路的影响

Effects of miR-375 on invasion,migration and JAK2/STAT3 signaling pathway of nasopharyngeal carcinoma cells

目的探讨微小RNA-375(miR-375)调节鼻咽癌细胞侵袭、迁移的作用及对Janus激酶2/信号转导与转录激活子3(JAK2/STAT3)信号通路的靶向调控。方法选取鼻咽癌CNE-1细胞并采用Lipofectamine 2000脂质体分别转染miR-375模拟物(miR-375组)和miR-375阴性对照(NC组),同时设不行转染的CNE-1细胞为对照组,采用实时荧光定量PCR(QPCR)检测转染48 h各组miR-375的表达情况;划痕实验和Transwell小室实验分别检测各组细胞的迁移能力和侵袭能力变化;Western blotting检测JAK2/STAT3信号通路中JAK2、STAT3蛋白的变化情况。结果 QPCR检测显示,miR-375在miR-375组的表达量为10.74±1.21,高于NC组的1.09±0.14和对照组的1.12±0.17,差异均有统计学意义(P<0.05);miR-375组的细胞迁移率为(18.22±1.78)%,低于对照组的(43.67±2.60)%和NC组的(49.70±1.31)%,差异均有统计学意义(P<0.05)。miR-375组的穿膜细胞数为(45.76±4.34)个,亦低于对照组的(144.98±3.65)个和NC组的(126.23±6.32)个,差异均有统计学意义(P<0.05);miR-375组JAK2、STAT3蛋白的表达水平分别为0.31±0.06和0.27±0.05,均低于对照组的0.54±0.05和0.41±0.06以及NC组的0.50±0.06和0.43±0.07(P<0.05),NC组和对照组以上指标的差异均无统计学意义(P>0.05)。结论上调miR-375表达可抑制CNE-1细胞的侵袭和迁移能力,可能与抑制JAK2/STAT3信号通路的激活有关。

Objective To explore the role of microRNA-375 （miR-375） on the invasion and migration of nasopharyngeal carcinoma cells and its targeting regulation of Janus activated kinase 2/sigual transducer and activator of transcription 3 （JAK2/STAT3） signaling pathway. Methods Nasopharyngeal carcinoma CNE-1 ceils were cultured and transfected with miR-375 mimics （miR-375 group） and miR-375 negative control （NC group） via Lipofectamine 2000 liposomes. CNE-1 cells without transfection were used as control group. The expression of miR-375 in each group at 48 h after transfection was detected by real-time quantitative PCR （QPCR）. The migration and invasion ability of the cells were detected by scratch test and Transwell test, respectively. Western blotting was used to detect the changes of JAK2 and STAT3 proteins in JAK2/STAT3 signaling pathway. Results The results of QPCR showed that the expression of miR-375 in miR-375 group was 10. 74±1.21, higher than 1.09±0. 14 of NC group and 1.12±0. 17 of control group （P〈0. 05）. The migration rate of miR-375 group was （ 18.22± 1.78） %, lower than （ 43.67 ±2. 60） % of control group and （ 49.70± 1.31 ） % of NC group （ P 〈0. 05）. The number of transmembrane cells in miR-375 group was 45.76±4. 34, also lower than 144. 98±3.65 of control group and 126. 23±6. 32 of NC group, and the difference was statistically significant （P〈0. 05 ）. The expression of JAK2 and STAT3 protein were 0. 31±0. 06 and 0. 27±0. 05 in miR-375 group, 0. 54±0. 05 and 0. 41±0. 06 in control group, and 0. 50±0. 06 and 0. 43±0. 07 in NC group. The expression of JAK2 and STAT3 in miR-375 group were lower than those in the other two groups （P〈0. 05）. There was no significant difference between NC group and control group （P〉0. 05）. Conclusion Up-regulation of miR-375 expression can inhibit the invasion and migration of CNE-1 cells. It may be related to the inhibition of JAK2/STAT3 signaling pathway.